RU2842948C1 - Modulating content of reducing sugars in a plant (inv) - Google Patents

Abstract

FIELD: biotechnology; genetic engineering.

SUBSTANCE: present invention describes a plant cell comprising (i) a polynucleotide comprising, consisting or substantially consisting of a sequence, characterized by at least 81 % sequence identity with SEQ ID NO: 5 (NtINV4-S) or at least 62 % sequence identity with SEQ ID NO: 7 (NtINV4-T); (ii) a polypeptide encoded by the polynucleotide indicated in (i); (iii) a polypeptide comprising, consisting or substantially consisting of a sequence having at least 85 % sequence identity with SEQ ID NO: 6 (NtINV4-S) or at least 85 % sequence identity with SEQ ID NO: 8 (NtINV4-T); or (iv) a construct, vector or expression vector comprising the isolated polynucleotide referred to in (i), wherein said plant cell provides at least one modification that provides modulation of (a) the expression or activity of the polynucleotide or (b) the expression or activity of the polypeptide, compared to those in the cell of a control plant in which the expression or activity of the polynucleotide or polypeptide has not been modified.

EFFECT: invention allows to produce a tobacco plant with new taste-and-flavour and organoleptical properties while maintaining commercially acceptable yields and characteristics.

18 cl, 5 dwg, 5 tbl, 9 ex

Description

FIELD OF TECHNOLOGY TO WHICH THE INVENTION RELATES

The present invention relates to plant cells and the like with modulated expression or activity of invertase (INV).

BACKGROUND OF THE INVENTION

To produce tobacco products, different types of tobacco are blended in varying ratios to create blends with specific flavor profiles. Flue-cured tobacco (e.g., Virginia) is the most widely grown tobacco and has a high sugar to nitrogen ratio but a limited flavor profile. Other types of tobacco, such as air-cured tobacco (e.g., Burley, Maryland, and Galpao) or fire-cured tobacco (e.g., Dark), offer alternative flavor profiles. These different flavor profiles are important in producing blended tobacco products.

Flavor properties are the result of specific flavor compounds or precursors of such compounds that are present in specific amounts in the tobacco plant. For example, altered sugar levels in cured tobacco can result in different perceptions of tobacco flavor and aroma. In aerosol and smoke, glucose and, to a lesser extent, fructose can form Amadori compounds via the Maillard reaction. This can result in bready, nutty, or popcorn-like flavor properties.

However, since the number of commercially available tobacco varieties is limited, this means that the ability to develop tobacco products with different flavour and aroma profiles is also limited. This is equally true for the production of reconstituted tobacco material, which is used in heated tobacco sticks in reduced risk products.

There remains a need in the art for improved capabilities to create tobacco that offers consumers new flavor and sensory experiences while maintaining commercially acceptable yields and characteristics. The present invention addresses these and other needs.

BRIEF DESCRIPTION OF THE INVENTION

Disclosed herein are INV polynucleotide and polypeptide sequences from Nicotiana tabacum . Although many different genes are thought to encode potential INVs in plants based on structural identity, the point at which these genes become active for their function as INVs in plants is generally unknown. In particular, very little is known about INV gene expression in tobacco, especially during curing. The present inventors have identified certain NtINV polynucleotides in plants that are functionally expressed during curing. Surprisingly, it has been found that modulating the expression of certain NtINV genes or the activity of the protein they encode can alter the pool of reducing sugars and sucrose formed during leaf curing. Surprisingly, it has been found that changes in certain INV genes can result in changes in the levels of reducing sugars and sucrose formed during leaf curing. Advantageously, this now provides the opportunity to create tobacco blends with new flavor and aroma properties. This may also result in a different flavor or sensory perception of the aerosol or smoke produced by heating the tobacco mixture. Likewise, liquid extracts obtained from tobacco may have a different flavor or sensory perception. Changing the balance of reducing sugars and sucrose may also affect the release of acrylamide into the aerosol and smoke.

This document describes several INV genomic polynucleotide sequences from Nicotiana tabacum , namely NtINV3-S (SEQ ID NO: 1), NtINV3-T (SEQ ID NO: 3), NtINV4-S (SEQ ID NO: 5), and NtINV4-T (SEQ ID NO: 7). Also described herein are several INV polypeptide sequences from Nicotiana tabacum , namely NtINV3-S (SEQ ID NO: 2), NtINV3-T (SEQ ID NO: 4), NtINV4-S (SEQ ID NO: 6), and NtINV4-T (SEQ ID NO: 8). In particular, NtINV4-S and NtINV4-T are shown to play a role in sugar metabolism during drying. During drying, flue-cured tobacco (e.g. Virginia) typically contains at least eight times more reducing sugars than air-cured tobacco (e.g. Burley), which is mainly due to its genetic predisposition to accumulate high levels of starch. After harvesting and during the aging process (yellowing phase), most of the starch is first converted to sucrose and then to reducing sugars, presumably with the help of INV.

The polypeptide sequences of NtINV4-S and NtINV4-T are very similar, sharing 96% identity. The situation is similar for the polypeptide sequences of NtINV3-S and NtINV3-T, which are also very similar, sharing 96% identity. Notably, the pair of NtINV4-S and NtINV4-T and the pair of NtINV3-S and NtINV3-T have low identity, only about 60%, suggesting different function or regulation. Surprisingly, only NtINV4-S and NtINV4-T are overexpressed during drying, while NtINV3-S and NtINV3-T are not overexpressed during drying. In certain embodiments, the expression or activity of NtINV3-S and NtINV3-T is not modulated. NtINV3-S and NtINV3-T are likely involved in other metabolic pathways and changes in their expression may result in a phenotype that may be unfavorable from an agronomic point of view (e.g., slow growth). Knowing which INV genes are overexpressed during desiccation advantageously allows selection of plants with changes in only the relevant genes and reduces potential negative effects on other metabolic processes.

Modifications of the expression or activity of one or more INVs can be combined with modifications of the expression or activity of one or more sucrose synthases (SUS) to providing additional modulation of sugar levels in dried leaves. Accordingly, a combination of modifications of INVs and SUS is disclosed. For example, reducing the expression or activity of one or more INVs as described herein can provide for an increase or decrease in glucose or fructose levels, or a combination thereof, and an increase in sucrose levels in a dried leaf. As a further example, increasing the expression or activity of one or more INVs as described herein can provide for an increase in glucose or fructose levels, or a combination thereof, and a decrease in sucrose levels in a dried leaf. As a further example, reducing the expression or activity of one or more INVs and one or more SUS as described herein can provide for a further decrease in glucose or fructose levels, or a combination thereof, in a dried leaf, compared to reducing the expression or activity of one or more INVs alone. As a further example, increasing the expression or activity of one or more INVs and one or more SUS, as described herein, can provide a further increase in glucose or fructose levels, or a combination thereof, in a dried leaf compared to increasing the expression or activity of one or more INVs alone.

This document disclosesNtSUS1-S(SEQ ID NO: 10), NtSUS1-T(SEQ ID NO: 12), NtSUS2-S(SEQ ID NO: 14),NtSUS2-T(SEQ ID NO: 16), NtSUS3-S(SEQ ID NO: 18), NtSUS3-T(SEQ ID NO:20), NtSUS4-S(SEQ ID NO: 22), NtSUS4-T(SEQ ID NO:24), NtSUS5-S(SEQ ID NO: 26), NtSUS5-T(SEQ ID NO:28), NtSUS6-S(SEQ ID NO: 30) AndNtSUS6-T(SEQ ID NO: 32). Also disclosed are the corresponding predicted polypeptide sequences for NtSUS1-S (SEQ ID NO: 11), NtSUS1-T (SEQ ID NO: 13), NtSUS2-S (SEQ ID NO: 15), NtSUS2-T (SEQ ID NO: 17), NtSUS3-S (SEQ ID NO: 19), NtSUS3-T (SEQ ID NO: 21), NtSUS4-S (SEQ ID NO: 23), NtSUS4-T (SEQ ID NO: 25), NtSUS5-S (SEQ ID NO: 27), NtSUS5-T (SEQ ID NO: 29), NtSUS6-S (SEQ ID NO: 31), and NtSUS6-T (SEQ ID NO: 33).NtSUS2-S,NtSUS2-T, NtSUS3-S,NtSUS3-T,NtSUS4-S And NtSUS4-Tmay play a role in sugar metabolism during drying. In particular,NtSUS2-S,NtSUS3-S,NtSUS3-T And NtSUS4-Smay play a role in sugar metabolism during drying.

In one aspect, a plant cell is provided, comprising (i) a polynucleotide comprising, consisting of, or essentially consisting of a sequence having at least 81% sequence identity to SEQ ID NO: 5 ( NtINV4-S ) or at least 62% sequence identity to SEQ ID NO: 7 ( NtINV4-T ); (ii) a polypeptide encoded by the polynucleotide set forth in (i); (iii) a polypeptide comprising, consisting of, or essentially consisting of a sequence having at least 84% or at least 85% sequence identity to SEQ ID NO: 6 (NtINV4-S) or at least 85% sequence identity to SEQ ID NO: 8 (NtINV4-T); or (iv) a construct, vector or expression vector comprising the isolated polynucleotide referred to in (i), wherein said plant cell comprises at least one modification that provides for modulation of (a) the expression or activity of the polynucleotide or (b) the expression or activity of a polypeptide derived from the polynucleotide, compared to that of a control plant cell in which the expression or activity of the polynucleotide or polypeptide has not been modified.

Preferably, the modulated expression or modulated activity provides for a modulation of the level of one or more reducing sugars in a dried leaf of a plant containing a plant cell, compared to the level of one or more reducing sugars in a dried leaf of a control plant containing a control plant cell, wherein preferably the reducing sugar is glucose or fructose or a combination thereof.

Preferably, the modulated expression or modulated activity also provides for modulation of the sucrose level in the dried plant leaf containing the plant cell.

Preferably, the dried leaf has reduced glucose levels of at least about 63% compared to a control dried leaf.

Preferably, the dried leaf has reduced fructose levels of at least about 43% compared to the control dried leaf. Preferably, the dried leaf has reduced glucose and fructose levels of at least about 63% and at least about 43%, respectively, compared to the control dried leaf.

Preferably, the dried leaf refers to a leaf obtained from the middle part of the plant.

Preferably, the effect on the phenotype of the plant containing the plant cell is negligible. For example, the phenotype of the plant may be unchanged.

Preferably, there is no variation in the total free amino acid content compared to the control plant containing the control plant cell.

Preferably, the at least one modification is at least one modification in the genome of the plant cell, or at least one modification in the construct, vector or expression vector, or at least one transgenic modification.

Preferably, at least one modification is a genetic mutation in the polynucleotide.

Preferably the plant is Nicotiana tabacum .

Preferably, the plant cell further comprises at least one modification in the polynucleotide.NtSUSor the polypeptide encoded by it, more preferably, where the polynucleotideNtSUSor the polypeptide encoded by it is selected from the group consisting ofNtSUS2-T,NtSUS3-S,NtSUS3-T,NtSUS4-S,NtSUS4-Tor a combination of two or more of them, more preferably, where polynucleotideNtSUSor the polypeptide encoded by it is selected from the group consisting ofNtSUS2-S,NtSUS3-S,NtSUS3-T And NtSUS4-Sor a combination of two or more of them.

Preferably, the plant cell contains at least one mutation in the polynucleotide(s)NtINV4or NtINV4 polypeptide(s) and at least one mutation in the polynucleotide(s)NtSUSor polypeptide(s), encoded by him/her.

In a further aspect, a plant or part thereof is provided comprising a plant cell as described herein.

A plant material, dried plant material or homogenized plant material originating or obtained from a plant or a part thereof are disclosed in a further aspect, wherein preferably the plant material is selected from the group consisting of biomass, seed, stem, flowers or leaves or a combination of two or more of them. The dried plant material may be selected from the group consisting of flue-dried plant material, sun-dried plant material or air-dried plant material or a combination of two or more of them.

In a further aspect, a tobacco product is provided comprising a plant cell, plant part or plant material.

In a further aspect, there is provided a method for producing a plant as described herein, comprising the steps of (a) obtaining a plant cell comprising at least one modification as described herein; and (b) propagating the plant cell to produce the plant.

Preferably, in step (a), at least one modification is introduced by genome editing; preferably, the genome editing methods are selected from CRISPR-mediated genome editing, zinc finger nuclease-mediated mutagenesis, chemical or radiation mutagenesis, homologous recombination, oligonucleotide-directed mutagenesis and meganuclease-mediated mutagenesis.

Preferably, in step (a), at least one modification is introduced using a polynucleotide to provide interference or by introducing at least one mutation or a combination thereof.

In a further aspect, a method is disclosed for producing dried plant material with an altered amount of reducing sugars compared to a control plant material, comprising the following steps: (a) obtaining a plant or part thereof or plant material described herein; (b) collecting plant material therefrom; and (c) drying the plant material.

In a further aspect, a method for producing a liquid tobacco extract is provided, wherein the method

comprises the steps of (a) obtaining a tobacco starting material from a plant or part thereof comprising a plant cell comprising at least one modification that provides for modulation of the expression or activity of NtINV as described herein; (b) heating the tobacco starting material at a suitable extraction temperature; (c) collecting volatile compounds released from the tobacco starting material during heating; and (d) combining the collected volatile compounds released from the tobacco starting material and forming a liquid tobacco extract.

In a further aspect, a method for producing a liquid tobacco extract is disclosed, the method comprising the steps of (a) obtaining a first tobacco starting material from a plant or a portion thereof comprising a plant cell in which the expression or activity of NtINV is modified as described herein; (b) obtaining a second tobacco starting material from a plant or a portion thereof comprising a plant cell in which the expression or activity of NtSUS is modified as described herein; (c) heating the first tobacco starting material at a first extraction temperature; (d) heating a second tobacco starting material at a second extraction temperature; (e) collecting volatile compounds released from the first tobacco starting materials and the second tobacco starting materials during the heating; and (f) combining the collected volatile compounds released from the first and second tobacco starting materials and forming a liquid tobacco extract from the combined volatile compounds.

In a further aspect, a liquid tobacco extract is disclosed, formed, obtained or obtainable by means of a method for producing a liquid tobacco extract as described herein.

In a further aspect, a plant cell is disclosed, comprising (i) a polynucleotide comprising, consisting of, or essentially consisting of a sequence having at least 60% sequence identity to SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, and SEQ ID NO: 7; a polynucleotide comprising, consisting of, or essentially consisting of a sequence having at least 50% sequence identity to SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8; (ii) a polypeptide encoded by the polynucleotide set forth in (i); (iii) a polypeptide comprising, consisting of, or essentially consisting of a sequence having at least 80% sequence identity to SEQ ID NO: 2 or SEQ ID NO: 4, 80% sequence identity to SEQ ID NO: 6, or SEQ ID NO: 8; (iv) a construct, vector or expression vector comprising the isolated polynucleotide referred to in (i), wherein said plant cell comprises at least one modification that provides for modulation of the expression or activity of the polynucleotide or polypeptide compared to that of a control plant cell in which the expression or activity of the polynucleotide or polypeptide has not been modified.

SOME ADVANTAGES

Primarily, changes in the sugar-amino acid balance in tobacco can affect the release of flavor compounds and acrylamide (a carcinogenic compound formed when glucose (fructose) reacts with asparagine) when heated in aerosol and smoke.

Preferably, reducing sugars are required for the proper production of a molded sheet of reconstituted tobacco material for heated tobacco sticks. The present invention may involve influencing the content and balance of sugars, thereby affecting the production of a molded sheet.

Preferably, non-genetically modified plants can be created that may be more acceptable to consumers.

Advantageously, the present invention is not limited to the use of plants obtained by EMS mutagenesis.

The present invention can be applied to various plant or crop varieties. Normally, senescent leaves (source leaves) produce sucrose as a carbon source and asparagine as an assimilated nitrogen resource for absorbent leaves and seeds. Therefore, sucrose and asparagine must be transported first from the parenchymatous (photosynthetic) senescent leaf cells to the phloem and then to the upper absorbent tissues. Manipulation of NtINV or the polypeptide encoded by it can thus affect the level of reducing sugars such as glucose and fructose, with little effect on free amino acids. Such an approach may provide the possibility of developing new tobacco varieties with lower glucose and fructose content and higher sucrose content.

Advantageously, the present invention can be combined with modulation of the expression of other genes, such as NtSUS or the polypeptide encoded therefrom, as described herein.

BRIEF DESCRIPTION OF GRAPHIC MATERIALS

Figure 1 shows a series of graphs depicting (A) the dynamics of reducing sugars (glucose and fructose) during flue-curing of Virginia tobacco; (B) the expression (Tobarray-Affymetrix) of NtINV3-S , NtINV3-T , NtINV4-S , and NtINV4-T during flue-curing of Virginia tobacco; and (C) the expression of the aging-associated gene SAG12 during flue-curing of Virginia tobacco.

Figure 2 shows a series of graphs showing (A) the dynamics of glucose levels; (B) the dynamics of fructose levels; and (C) the dynamics of sucrose levels during air-curing of dark tobacco. The data were obtained from Metabolon metabolomics analyses (no units available).

Figure 3 shows a graph showing the results regarding the silencing of NtINV4-S and NtINV4-T using the GATEWAY vector and the measurement of the expression of NtINV4-S and NtINV4-T in Virginia tobacco leaf after 48 h of drying (qPCR). T0-INV4 is the generated transgenic line and CT0-INV4 are the corresponding control lines.

Figure 4 shows a series of graphs showing the contents of (A) glucose, (B) fructose, and (C) sucrose in dried leaves silenced with 35S:INV4-RNAi (INV4-T0) and dried control leaves (CT0) (CT0, n=4 and T0, n=4). Box plots are shown, as well as statistical analysis by T-test.

Onfigure 5presented A bar chart showing for each variety the reducing sugar content after picking (mature leaves), after two days of drying (48 hours drying), and at the end of drying in Burley, Virginia, and Oriental tobaccos.

DETAILED DESCRIPTION

The section headings used in this disclosure are for organizational purposes and are not intended to be limiting.

1. Definitions

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. In case of conflict, this document, including definitions, will control. Preferred methods and materials are described below, although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. The materials, methods, and examples disclosed herein are illustrative only and are not intended to be limiting.

The terms “provide for,” “include,” “having,” “has,” “may,” “contain,” and variations thereof are intended to be open-ended transitional phrases, terms, or words that do not exclude the possibility of additional actions or structures.

Singular forms include references to the plural unless the context clearly indicates otherwise.

The present invention provides other embodiments "comprising," "consisting of," and "consisting essentially of" the embodiments or elements presented herein, whether or not expressly stated.

When numerical ranges are set forth in this document, each intermediate number therein is explicitly provided with the same degree of precision. For example, in the case of the range 6-9, in addition to 6 and 9, the numbers 7 and 8 are provided, and in the case of the range 6.0-7.0, the numbers 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, and 7.0 are explicitly provided.

The following terms used throughout this description and claims have the following meanings.

The terms "coding sequence" or "coding polynucleotide" mean the nucleotides (RNA or DNA molecules) that make up a polynucleotide that codes for a polypeptide. The coding sequence may further comprise initiation and termination signals operably linked to regulatory elements, including a promoter and a polyadenylation signal, capable of directing expression in cells of an individual or mammal to which the polynucleotide is administered. The coding sequence may be codon-optimized.

The terms "complementary sequence" or "complementary" can refer to Watson-Crick formation (e.g. A-T/U and C-G) or Hoogsteen pairing between nucleotides or nucleotide analogues. "Complementarity" refers to the property shared by two polynucleotides that when they are aligned antiparallel to each other, the nucleotide bases at each position are complementary to each other.

The term "construct" refers to a double-stranded fragment of a recombinant polynucleotide comprising one or more polynucleotides. The construct comprises a "template strand" whose bases are paired with a complementary "sense or coding strand". The construct may be inserted into a vector in two possible orientations: either in the same (or sense) orientation or in the opposite (or antisense) orientation to the orientation of a promoter located in a vector, such as an expression vector.

The term "control" in the context of a control plant or control plant cells means a plant or plant cells in which the expression, function or activity of one or more genes or polypeptides has not been modified (e.g. increased or decreased) and can therefore provide a comparison with a plant in which the expression, function or activity of one or more of the same genes or polypeptides has been modified. "Control plant" means a plant that is substantially equivalent to a test plant or modified plant in all respects except the respects being tested. For example, in the case of a plant into which a polynucleotide has been introduced, the control plant is an equivalent plant into which such polynucleotide has not been introduced. The control plant may be an equivalent plant into which a control polynucleotide has been introduced. In such cases, the control polynucleotide is a polynucleotide for which the phenotypic effect it causes on the plant is expected to be negligible or absent. The control plant may contain an empty vector. The control plant may correspond to a wild-type plant. The control plant may be a null segregant, in which case the T1 segregant no longer contains the transgene.

The term "reduction" or "reduced" refers to a reduction of from about 10% to about 99%, or a reduction of at least 10%, at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 90%, at least 95%, at least 98%, at least 99%, or at least 100%, or at least 150%, or at least 200%, or more, in an amount or function, such as polypeptide function, transcriptional function, or polypeptide expression. The term "reduced" or the expression "reduced amount" may refer to an amount or feature that is less than what would be found in a plant or product of the same plant variety processed in the same way that has not been modified. Thus, in some cases, a wild-type plant of the same variety that has been processed in the same way is used as a control by which to measure whether a reduction in amount has been achieved.

The terms "donor DNA" or "donor template" refer to a fragment or molecule of double-stranded DNA that contains at least a portion of a gene of interest. Donor DNA may encode a functional polypeptide.

The term "endogenous gene or polypeptide" refers to a gene or polypeptide that originates from the genome of an organism and has not undergone change, such as loss, gain, or replacement of genetic material. An endogenous gene undergoes normal gene transfer and gene expression. An endogenous polypeptide undergoes normal expression.

The term "enhancer sequences" refers to sequences that can mediate increased gene expression. These sequences can be located upstream, within introns, or downstream of the transcribed region. The transcribed region consists of exons and introns located between them, from the promoter to the transcription termination region. Increased gene expression can occur through a variety of mechanisms, including increased transcription efficiency, stabilization of mature mRNA, and increased translation.

The term "expression" refers to the production of a functional product. For example, expression of a polynucleotide fragment may refer to transcription of the polynucleotide fragment (e.g., transcription resulting in mRNA or functional RNA) or translation of mRNA to produce a precursor polypeptide or a mature polypeptide, or a combination thereof.

"Overexpression" refers to the production of a gene product in transgenic organisms at levels that exceed the levels of production in a null-segregant (or non-transgenic) organism from the same experiment.

The term "functional" describes a polypeptide that has a biological function or activity. The term "functional gene" refers to a gene that is transcribed to produce mRNA that is translated to produce a functional or active polypeptide.

The term "genetic construct" refers to DNA or RNA molecules that contain a polynucleotide that encodes a polypeptide. The coding sequence may contain initiation and termination signals operably linked to regulatory elements, including a promoter and a polyadenylation signal, capable of directing expression.

"Genome editing" generally refers to the process by which a change is made to the genomic nucleic acid in a cell. This may be done, for example, by deleting, inserting, or replacing one or more nucleotides in the genomic nucleic acid. Endonucleases can be used to create specific breaks or single-strand breaks at specific locations in the genome, and are further described herein.

The terms "homology" or "similarity" refer to the degree of similarity between the sequences of two polypeptides or two polynucleotide molecules compared by sequence alignment. The degree of homology between two individual polynucleotides being compared is a function of the number of identical or matching nucleotides at the positions being compared. Homology or similarity can be determined over the entire length of the sequence in question.

The terms "identical" or "identity" in the context of two or more polynucleotides or polypeptides mean that the sequences are characterized by having a specified percentage of residues that are the same within a specified region. The percentage can be calculated by optimally aligning the two sequences, comparing the two sequences within a specified region, determining the number of positions at which identical residues occur in both sequences to yield the number of matching positions, dividing the number of matching positions by the total number of positions within the specified region, and multiplying the result by 100 to yield the percentage sequence identity. In cases where the two sequences are of different lengths or the alignment creates one or more asymmetric ends and the specified comparison region includes only one sequence, the residues of the single sequence are included in the denominator rather than the numerator in the calculation. Thymine (T) and uracil (U) can be considered equivalent when comparing DNA and RNA. Identity can be determined manually or using a computer algorithm for working with sequences, such as ClustalW, ClustalX, BLAST, FASTA, or the Smith-Waterman algorithm. Suitable parameters for ClustalW are as follows. For polynucleotide alignments: gap-open penalty=15.0, gap-extension penalty=6.66, and matrix=identity. For polypeptide alignments: gap-open penalty=10.0, gap-extension penalty=0.2, and matrix=Gonnet. For DNA and protein alignments: ENDGAP=-1 and GAPDIST=4.

The terms "increase" or "increased" refer to an increase of from about 10% to about 99%, or an increase of at least 10%, at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 90%, at least 95%, at least 98%, at least 99%, at least 100%, at least 150%, or at least 200% or more, in quantity or function or activity, such as, but not limited to, one or more of a polypeptide function or activity, a transcriptional function, or a polypeptide activity and expression. The term "increased" or the expression "increased amount" may refer to an amount or function or activity in a plant or plant product that is greater than that found in a plant or plant product of the same plant variety treated in the same way that has not been modified. Thus, in some cases, a wild-type plant of the same variety as a plant that has been treated in the same way is used as a control by which to measure whether an increase in amount has been achieved.

The term "inhibit" or "inhibited" refers to a reduction of from about 98% to about 100%, or a reduction of at least 98%, at least 99%, and in particular 100%, in an amount or function or activity, such as, but not limited to, one or more of a polypeptide function or activity, a transcriptional function, or the activity and expression of a polypeptide.

The term "introduced" means the delivery of a polynucleotide (e.g., a construct) or a polypeptide into a cell. The term "introduced" includes reference to the incorporation of a polynucleotide into a eukaryotic cell, where the polynucleotide may be incorporated into the cell's genome, and also includes reference to the transient introduction of a polynucleotide or polypeptide into a cell. The term "introduced" includes references to stable or transient transformation methods, as well as sexual mating. Thus, the term "introduced" when applied to the insertion of a polynucleotide (e.g., a recombinant construct/expression construct) into a cell means "transfection" or "transformation" or "transduction" and includes reference to the insertion of a polynucleotide into a eukaryotic cell, where the polynucleotide may be incorporated into the cell's genome (e.g., chromosomal, plasmid, plastid, or mitochondrial DNA), converted into an autonomous replicon, or expressed transiently (e.g., transfected mRNA).

The terms "isolated" or "purified" refer to a material that is substantially or substantially free of components that normally accompany it as found in its native state. Typically, purity and homogeneity are determined by analytical chemistry techniques such as polyacrylamide gel electrophoresis or high performance liquid chromatography. A polypeptide that is the predominant molecule present in a preparation is substantially purified. In particular, an isolated polynucleotide is separated from open reading frames that flank the desired gene and encode polypeptides other than the desired polypeptide. The term "purified" means that the polynucleotide or polypeptide yields substantially a single band in an electrophoretic gel. In particular, this means that the polynucleotide or polypeptide is at least 85% pure, more preferably at least 95% pure, and most preferably at least 99% pure. The isolated polynucleotides may be purified from the host cell in which they naturally occur. Conventional nucleic acid purification techniques known to those skilled in the art may be used to obtain the isolated polynucleotides. The term also includes recombinant polynucleotides and chemically synthesized polynucleotides.

"Liquid tobacco extract" describes the direct product of an extraction process performed on a tobacco starting material. The extraction method for producing a liquid tobacco extract may include heating the tobacco starting material under specific heating conditions and collecting the volatile compounds formed. The liquid tobacco extract may contain a mixture of compounds originating from the tobacco starting material and removed during the extraction procedure, typically in combination with a liquid carrier or solvent.

The terms "modulate" or "modulation" refer to providing or facilitating a qualitative or quantitative change, adjustment or modification of a process, pathway, function or activity of interest. Without limitation, such a change, adjustment or modification may be an increase or decrease in the level of the corresponding process, pathway, function or activity of interest. For example, gene expression or polypeptide expression or polypeptide function or activity may be modulated. Typically, the relative change, adjustment or modification is determined by comparison with a control.

The term "non-naturally occurring" describes an object, such as a polynucleotide, a genetic mutation, a polypeptide, a plant, a plant cell, and plant material, that is not naturally formed or does not exist in nature. Such non-naturally occurring objects or artificial objects can be created, synthesized, initiated, modified, interfered with, or manipulated by the methods described herein or known in the art. Such non-naturally occurring objects or artificial objects can be created, synthesized, initiated, modified, interfered with, or manipulated by a human. Thus, as an example, a non-naturally occurring plant, non-naturally occurring plant cell or non-naturally occurring plant material can be created using traditional plant breeding techniques such as backcrossing or using gene manipulation technologies such as antisense RNA, interfering RNA, meganuclease, etc. As a further example, a non-naturally occurring plant, non-naturally occurring plant cell or non-naturally occurring plant material can be created by introgression or by transferring one or more genetic mutations (e.g., one or more polymorphisms) from a first plant or plant cell to a second plant or plant cell (which itself may be naturally occurring), such that the resulting plant, plant cell or plant material or progeny thereof comprises a genetic structure (e.g., a genome, chromosome or segment thereof) that is not naturally occurring or that does not exist in nature. The resulting plant, plant cell or plant material is thus artificial or non-naturally occurring. Accordingly, an artificial or non-naturally occurring plant or plant cell can be created by modifying a genetic sequence in a first naturally occurring plant or plant cell, even if the resulting genetic sequence is naturally occurring in a second plant or plant cell that has a genetic background different from that of the first plant or plant cell. In certain embodiments, the mutation is not a naturally occurring mutation that naturally exists in a polynucleotide or polypeptide, such as a gene or polypeptide. Differences in genetic background can be detected by phenotypic differences or by molecular biology techniques known in the art, such as sequencing a polynucleotide, determining the presence or absence of genetic markers (e.g., microsatellite RNA markers).

The terms "oligonucleotide" or "polynucleotide" mean at least two nucleotides covalently linked together. The description of a single strand also defines the sequence of the complementary strand. Thus, a polynucleotide also encompasses a strand complementary to the described single strand. Many variants of a polynucleotide can be used for the same purpose as the specified polynucleotide. Thus, a polynucleotide also encompasses substantially identical polynucleotides and their complementary sequences. A single strand is a probe that can hybridize to a given sequence under stringent hybridization conditions. Thus, a polynucleotide also encompasses a probe that hybridizes under stringent hybridization conditions. Polynucleotides may be single-stranded or double-stranded, or may contain portions of both double-stranded and single-stranded sequences. A polynucleotide may be DNA, either genomic or cDNA, RNA, or a hybrid molecule, wherein a polynucleotide may contain combinations of deoxyribo- and ribonucleotides, as well as combinations of bases, including uracil, adenine, thymine, cytosine, guanine, inosine, xanthine, hypoxanthine, isocytosine, and isoguanine. Polynucleotides may be produced by chemical synthetic methods or by recombinant methods.

The specificity of single-stranded DNA for hybridization to complementary fragments is determined by the "stringency" of the reaction conditions (Sambrook et al ., Molecular Cloning and Laboratory Manual, Second Ed., Cold Spring Harbor (1989)). For hybridization under "stringent conditions," hybridization protocols are described in which polynucleotides that are at least 60% homologous to each other remain hybridized. Typically, stringent conditions are chosen such that the temperature is approximately 5ºC lower than the melting point (Tm) temperature for a particular sequence at specified ionic strength and pH. Tm is the temperature (at specified ionic strength, pH, and polynucleotide concentration) at which 50% of the probes complementary to a given sequence hybridize to that sequence at equilibrium. Since the sequences are usually present in excess, 50% of the probes are occupied at equilibrium at Tm.

Stringent conditions typically include: (1) low ionic strength and high temperature washes, e.g. 15 mM sodium chloride, 1.5 mM sodium citrate, 0.1% sodium dodecyl sulfate at 50ºC; (2) the presence of a denaturing agent during hybridization, e.g. 50% (v/v) formamide, 0.1% bovine serum albumin, 0.1% Ficoll, 0.1% polyvinylpyrrolidone, 50 mM sodium phosphate buffer (750 mM sodium chloride, 75 mM sodium citrate; pH 6.5) at 42ºC or (3) the presence of 50% formamide. Typically, washes also include 5x SSC (0.75 M NaCl, 75 mM sodium citrate), 50 mM sodium phosphate (pH 6.8), 0.1% sodium pyrophosphate, 5x Denhardt's solution, sonicated salmon milt DNA (50 μg/ml), 0.1% SDS, and 10% dextran sulfate at 42ºC with a 42ºC wash in 0.2x SSC (sodium chloride/sodium citrate) and 50% formamide at 55ºC followed by a high stringency wash of 0.1x SSC containing EDTA at 55ºC. Preferably, the conditions are such that sequences that are at least about 65%, 70%, 75%, 85%, 90%, 95%, 98%, or 99% homologous to each other generally remain hybridized to each other.

"Moderate stringency conditions" use wash solutions and hybridization conditions that are less stringent, such that the polynucleotide hybridizes to the entire polynucleotide of interest, its fragments, derivatives, or analogs. One example is hybridization in 6x SSC, 5x Denhardt's solution, 0.5% SDS, and 100 μg/ml denatured salmon milt DNA at 55ºC, followed by one or more washes in 1x SSC, 0.1% SDS at 37ºC. Temperature, ionic strength, etc., can be adjusted to accommodate experimental factors such as probe length. Other moderately stringent conditions have been described (see Ausubel et al., Current Protocols in Molecular Biology, Volumes 1–3, John Wiley & Sons, Inc., Hoboken, N.J. (1993); Kriegler, Gene Transfer and Expression: A Laboratory Manual, Stockton Press, New York, N.Y. (1990); Perbal, A Practical Guide to Molecular Cloning, 2nd edition, John Wiley & Sons, New York, N.Y. (1988)).

Under "low stringency conditions" washing solutions and hybridization conditions are used that are less stringent than those under moderate stringency, such that the polynucleotide hybridizes to the entire polynucleotide in question, its fragments, derivatives, or analogs. A non-limiting example of low stringency hybridization conditions is hybridization in 35% formamide, 5x SSC, 50 mM Tris-HCl (pH 7.5), 5 mM EDTA, 0.02% PVP, 0.02% Ficoll, 0.2% BSA, 100 μg/ml denatured salmon milt DNA, 10% (w/v) dextran sulfate at 40ºC followed by one or more washes in 2x SSC, 25 mM Tris-HCl (pH 7.4), 5 mM EDTA, and 0.1% SDS at 50ºC. Other low stringency conditions, such as those for interspecies hybridization variants, have been well described (see Ausubel et al., 1993; Kriegler, 1990).

The term "operably linked" means that the expression of a gene is under the control of a promoter to which it is spatially linked. A promoter may be located 5' (upstream) or 3' (downstream) of the gene that is under its control. The distance between a promoter and a gene may be approximately the same as the distance between the promoter and the gene that it controls in the gene from which the promoter is derived. As is known in the art, this distance can be varied in a coordinated manner without loss of promoter function. The term "operably linked" refers to the association of polynucleotide fragments in one fragment such that the function of one is regulated by the other. For example, a promoter is operably linked to a polynucleotide fragment if it is capable of regulating the transcription of that polynucleotide fragment.

The term "plant" refers to any plant at any stage of its life cycle or development and its progeny. In one embodiment, the plant is a tobacco plant, which refers to a plant belonging to the genus Nicotiana. The term includes reference to whole plants, plant organs, plant tissues, plant propagules, plant seeds, and plant cells and their progeny. Plant cells include, but are not limited to, cells from seeds, suspension cultures, embryos, meristematic regions, callus tissue, leaves, roots, shoots, gametophytes, sporophytes, pollen, and microspores. Suitable tobacco plant species, cultivars, hybrids, and varieties are described herein.

"Plant material" includes leaf, root, sepal, root tip, petal, flower, shoot, stem, seed, and petiole. Plant material may be viable or non-viable plant material.

The terms "polynucleotide," "polynucleotide sequence," or "polynucleotide fragment" are used interchangeably herein and refer to a polymer of RNA or DNA that is single- or double-stranded and optionally contains synthetic, non-natural, or corrected nucleotide bases. The polynucleotides of the present invention are provided in the attached sequence listing.

The terms "polypeptide" or "polypeptide sequence" refer to a polymer of amino acids in which one or more amino acid residues are an artificial chemical analog of the corresponding naturally occurring amino acid, as well as to naturally occurring polymers of amino acids. The terms also include modifications including, but not limited to, glycosylation, lipid addition, sulfation, gamma-carboxylation of glutamic acid residues, hydroxylation, and ADP-ribosylation. The polypeptides of the present invention are provided in the attached sequence listing.

The term "promoter" means a synthetic or naturally occurring molecule that is capable of promoting, activating or enhancing the expression of a polynucleotide in a cell. The term refers to an element/sequence of a polynucleotide, typically located upstream of and operably linked to a portion of a double-stranded polynucleotide. Promoters may be derived entirely from regions near a native gene of interest, or may consist of different elements derived from different native promoters or segments of a synthetic polynucleotide. A promoter may contain one or more specific transcriptional regulatory sequences to further enhance expression, or to alter spatial expression, or to alter temporal expression. A promoter may also contain distal enhancer or repressor elements, which may be located up to several thousand base pairs from the transcription start site. A promoter may be derived from sources including viruses, bacteria, fungi, plants, insects and animals. A promoter may regulate the expression of a gene component constitutively or differentially with respect to the cell, tissue, or organ in which expression occurs, or with respect to the developmental stage at which expression occurs, or in response to external stimuli such as physiological stress, pathogens, metal ions, or inducing agents.

The terms "tissue-specific promoter" and "tissue-preferred promoter" as used interchangeably herein refer to a promoter that is expressed predominantly, but not necessarily exclusively, in one organ or tissue, but may also be expressed in one particular cell. The term "developmentally regulated promoter" refers to a promoter whose function is determined by developmental events. The term "constitutive promoter" refers to a promoter that causes expression of a gene in most cell types under most circumstances. An "inducible promoter" causes selective expression of an operably linked DNA sequence in response to the presence of endogenous or exogenous stimuli, such as chemical compounds (chemical inducers), or in response to environmental, hormonal, chemical, or developmental signals, or a combination of two or more thereof. Examples of inducible or regulated promoters include promoters regulated by light, heat, stress, flooding or drought, pathogens, phytohormones, wounding, or chemicals such as ethanol, jasmonate, salicylic acid, or antidotes.

The term "recombinant" refers to an artificial combination of two otherwise separated segments of a sequence, obtained, for example, by chemical synthesis or by manipulation of isolated segments of polynucleotides using genetic engineering techniques. The term also includes reference to a cell or vector that has been modified by the introduction of a heterologous polynucleotide, or a cell derived from a cell so modified, but does not encompass adjustment of the cell or vector by naturally occurring events (e.g., by spontaneous mutation, natural transformation, or transduction, or transposition), such as those that occur without deliberate human intervention.

The term "recombinant construct" refers to a combination of polynucleotides that are not normally found together in nature. Accordingly, a recombinant construct may contain regulatory sequences and coding sequences obtained from different sources, or regulatory sequences and coding sequences obtained from the same source but arranged differently from what is normally found in nature. A recombinant construct may be a recombinant DNA construct.

The terms "regulatory sequences" and "regulatory elements" are used interchangeably herein to refer to polynucleotide sequences located upstream (5' non-coding sequences), within, or downstream (3' non-coding sequences) of a coding sequence that affect transcription, processing, or stability of RNA or translation of an associated coding sequence. Regulatory sequences include promoters, translational leader sequences, introns, and polyadenylation recognition sequences. The terms "regulatory sequence" and "regulatory element" are used interchangeably herein.

The term "tobacco" is used in a collective sense to refer to agricultural tobacco crops (e.g., a plurality of field-grown tobacco plants and tobacco grown in a manner other than hydroponically), tobacco plants, and parts thereof, including but not limited to roots, stems, leaves, flowers, and seeds, prepared or obtained as described herein. It is understood that "tobacco" includes Nicotiana tabacum plants and products thereof.

The term "tobacco products" refers to consumer tobacco products, including, but not limited to, smoking materials (e.g., cigarettes, cigars, and pipe tobacco), snuff, chewing tobacco, chewing gum, and lozenges, as well as components, materials, and ingredients for the manufacture of consumer tobacco products. These tobacco products are preferably manufactured from tobacco leaves and stems that have been harvested from the tobacco plant and cut, cured, dried, or fermented in accordance with conventional tobacco production techniques.

The terms "transcription terminator", "termination sequences" or "terminator" refer to DNA sequences located downstream of the coding sequence, including recognition sequences of polyadenylation and other sequences encoding regulatory signals capable of affecting mRNA processing or gene expression. The polyadenylation signal is typically characterized by the addition of polyadenylic acid tracts to the 3' end of the mRNA precursor.

The term "transgenic" refers to any cell, cell line, callus, tissue, plant part, or plant whose genome has been adjusted by the presence of a heterologous polynucleotide such as a recombinant construct, including the original transgenic event as well as those obtained by sexual crossing or asexual propagation procedures from the original transgenic event. The term does not cover adjustment of the genome (chromosomal or extrachromosomal) by conventional plant breeding techniques or by naturally occurring events such as random cross-pollination, infection by a non-recombinant virus, transformation by non-recombinant bacteria, non-recombinant transposition, or spontaneous mutation.

The term "transgenic plant" refers to a plant that contains one or more heterologous polynucleotides in its genome, i.e., a plant that contains recombinant genetic material not normally found in the plant and that has been introduced into the plant in question (or into the ancestors of the plant) by human manipulation. For example, a heterologous polynucleotide may be stably integrated into the genome in such a way that the polynucleotide is passed on to subsequent generations. The heterologous polynucleotide may be integrated into the genome alone or as part of a recombinant construct. Commercial development of genetically improved germplasm has also advanced to the stage of introducing multiple traits into crop plants, often referred to as the gene pyramiding approach. In this approach, multiple genes conferring different characteristics of interest may be introduced into the plant. Gene pyramiding may be accomplished in a variety of ways, including, but not limited to, cotransformation, retransformation, and crossing lines with different transgenes. Thus, a plant grown from a plant cell into which recombinant DNA has been introduced by transformation is a transgenic plant, as are all progeny of that plant that contain the introduced transgene (whether produced sexually or asexually). It is understood that the term "transgenic plant" covers the entire plant or tree and parts of the plant or tree, such as grains, seeds, flowers, leaves, roots, fruits, pollen, stems, etc. Each heterologous polynucleotide may impart a distinct trait to the transgenic plant.

The term "transgene" refers to a gene or genetic material containing a gene sequence that has been isolated from one organism and introduced into another organism. This non-native segment of DNA may retain the ability to produce RNA or a polypeptide in the transgenic organism, or it may provide a correction to the normal function of the transgenic organism's genetic code.

The term "variant" with respect to a polynucleotide means: (i) a portion or fragment of a polynucleotide; (ii) a sequence complementary to the polynucleotide or a portion thereof; (iii) a polynucleotide substantially identical to the reference polynucleotide or a sequence complementary thereto; or (iv) a polynucleotide that hybridizes under stringent conditions to the reference polynucleotide, a sequence complementary thereto, or a polynucleotide substantially identical thereto.

The term "variant" when used with respect to a peptide or polypeptide means a peptide or polypeptide that differs in sequence by insertion, deletion, or conservative substitution of amino acids but retains at least one biological function or activity. A variant may also mean a polypeptide that retains at least one biological function or activity. A conservative amino acid substitution, i.e., replacing an amino acid with another amino acid with similar properties (e.g., degree of hydrophilicity and distribution of charged sites), is understood in the art to generally involve a minor change.

The term cultivar refers to a population of plants that have consistent characteristics that distinguish them from other plants of the same species. Although a cultivar has one or more distinguishing characteristics, it is further characterized by very little overall variation between individuals within the cultivar. A cultivar is often sold commercially.

The term "vector" refers to a polynucleotide delivery vehicle that contains a combination of polynucleotide components to provide for the transport of polynucleotides, polynucleotide constructs and polynucleotide conjugates, etc. The vector may be a viral vector, a bacteriophage, a bacterial artificial chromosome or a yeast artificial chromosome. The vector may be a DNA or RNA vector. Suitable vectors include episomes capable of extrachromosomal replication, such as circular plasmids of double-stranded nucleotide sequence; linearized plasmids of double-stranded nucleotide sequence; and other vectors of any origin. An "expression vector" is a polynucleotide delivery vehicle that contains a combination of polynucleotide components to provide for the expression of polynucleotide(s), polynucleotide constructs and polynucleotide conjugates, etc. Suitable expression vectors include episomes capable of extrachromosomal replication, such as circular plasmids of double-stranded nucleotide sequence; linearized plasmids from a double-stranded nucleotide sequence and other functionally equivalent expression vectors of any origin. The expression vector comprises at least a promoter located upstream of and operably linked to a polynucleotide, polynucleotide constructs or a polynucleotide conjugate as defined below.

Unless otherwise defined herein, scientific and technical terms used in connection with the present disclosure shall have meanings that are commonly understood by those of ordinary skill in the art. For example, any nomenclature systems and techniques used in connection with cell or tissue culture, molecular biology, immunology, microbiology, genetics, and polypeptide and polynucleotide chemistry and hybridization that are described herein are well known and commonly used in the art. The meaning and scope of terms should be clear, but in the event of any hidden ambiguity, the definitions provided herein take precedence over any dictionary or unrelated definition. Furthermore, unless the context otherwise requires, singular terms shall include the plural, and plural terms shall include the singular.

2. Polynucleotides

An isolated polynucleotide is disclosed that comprises, consists of, or essentially consists of a sequence having at least 60% sequence identity to any of the sequences described herein, including any of the polynucleotides shown in the sequence listing. Preferably, the isolated polynucleotide comprises, consists of, or essentially consists of a sequence having at least 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 75%, 80%, 85%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity thereto.

Preferably, the polynucleotide(s) described herein encode(s) an active polypeptide that has at least about 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, 100% or more of the function or activity of the polypeptide(s) shown in the sequence listing.

In another embodiment, an isolated NtINV polynucleotide is provided, comprising, consisting of, or consisting essentially of a polynucleotide having at least 60% sequence identity to SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, or SEQ ID NO: 9.

In another embodiment, an isolated NtSUS polynucleotide is provided, comprising, consisting of, or consisting essentially of a polynucleotide having at least 60% sequence identity to SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO: 22, or SEQ ID NO: 24; SEQ ID NO: 26 , SEQ ID NO: 28, SEQ ID NO: 30, or SEQ ID NO: 32.

Preferably, the isolated polynucleotide comprises, consists of, or consists essentially of a sequence having at least about 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 75%, 80%, 85%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, or 100% sequence identity to SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7 or SEQ ID NO: 9.

Preferably, the isolated polynucleotide comprises, consists of, or consists essentially of a sequence having at least about 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 75%, 80%, 85%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, or 100% sequence identity to SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO: 22 or SEQ ID NO: 24; SEQ ID NO: 26 , SEQ ID NO: 28, SEQ ID NO: 30 or SEQ ID NO: 32.

Preferably, the isolated polynucleotide comprises, consists of, or essentially consists of a sequence having at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, or 100% sequence identity to SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, or SEQ ID NO: 9.

Preferably, the isolated polynucleotide comprises, consists of, or consists essentially of a sequence having at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, or 100% sequence identity to SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO: 22 or SEQ ID NO: 24; SEQ ID NO: 26 , SEQ ID NO: 28, SEQ ID NO: 30 or SEQ ID NO: 32.

Preferably, the isolated polynucleotide comprises, consists of, or essentially consists of a sequence having at least about 95%, 96%, 97%, 98%, 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, or 100% sequence identity to SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, or SEQ ID NO: 9.

Preferably, the isolated polynucleotide comprises, consists of, or essentially consists of a sequence having at least about 95%, 96%, 97%, 98%, 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, or 100% sequence identity to SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO: 22, or SEQ ID NO: 24; SEQ ID NO: 26 , SEQ ID NO: 28, SEQ ID NO: 30, or SEQ ID NO: 32.

In another embodiment, polynucleotides are provided that comprise, consist of, or consist essentially of polynucleotides with a substantial degree of homology (i.e., sequence similarity) or a substantial degree of identity to SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, or SEQ ID NO: 9.

In another embodiment, polynucleotides are provided that comprise, consist of, or essentially consist of polynucleotides with a substantial degree of homology (i.e., sequence similarity) or a substantial degree of identity to SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO: 22, or SEQ ID NO: 24; SEQ ID NO: 26 , SEQ ID NO: 28, SEQ ID NO: 30, or SEQ ID NO: 32.

In another embodiment, fragments of SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, or SEQ ID NO: 9 are provided with a substantial degree of homology (i.e., sequence similarity) or a substantial degree of identity thereto that are characterized by at least about 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 75%, 80%, 85%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9% or 100% sequence identity to the corresponding fragments of SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7 or SEQ ID NO: 9.

In another embodiment, fragments of SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO: 22, or SEQ ID NO: 24 are provided; SEQ ID NO: 26 , SEQ ID NO: 28, SEQ ID NO: 30 or SEQ ID NO: 32 with a substantial degree of homology (i.e., sequence similarity) or with a substantial degree of identity thereto, which are characterized by at least about 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 75%, 80%, 85%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9% or 100% sequence identity to the corresponding fragments of SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO: 22 or SEQ ID NO: 24; SEQ ID NO: 26 , SEQ ID NO: 28, SEQ ID NO: 30 or SEQ ID NO: 32.

In another embodiment, polynucleotides are provided that provide a sufficient or substantial degree of identity or similarity to SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, or SEQ ID NO: 9, which encode a polypeptide that functions as an INV.

In another embodiment, polynucleotides are provided that provide a sufficient or substantial degree of identity or similarity to SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO: 22 or SEQ ID NO: 24; SEQ ID NO: 26 , SEQ ID NO: 28, SEQ ID NO: 30 or SEQ ID NO: 32, which encode a polypeptide that functions as SUS.

In another embodiment, a polymer is provided that is a polynucleotide that comprises, consists of, or essentially consists of a polynucleotide designated herein as SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, or SEQ ID NO: 9.

In another embodiment, a polymer is provided that is a polynucleotide that comprises, consists of, or essentially consists of a polynucleotide designated herein as SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO: 22, or SEQ ID NO: 24; SEQ ID NO: 26 , SEQ ID NO: 28, SEQ ID NO: 30, or SEQ ID NO: 32.

The polynucleotides described herein preferably encode members of the INV family or the SUS family, which are characterized by INV activity or SUS activity, respectively.

A polynucleotide may comprise a polymer of nucleotides, which may be unmodified or modified deoxyribonucleic acid (DNA) or ribonucleic acid (RNA). Accordingly, a polynucleotide may be, without limitation, genomic DNA, complementary DNA (cDNA), mRNA, or antisense RNA, or fragment(s) thereof. In addition, a polynucleotide may be single-stranded or double-stranded DNA, DNA that is a mixture of single-stranded and double-stranded regions, a hybrid molecule containing DNA and RNA, or a hybrid molecule with a mixture of single-stranded and double-stranded regions, or fragment(s) thereof. Additionally, a polynucleotide may be composed of triple-stranded regions containing DNA, RNA, or both, or fragment(s) thereof. A polynucleotide may contain one or more modified bases, such as phosphorothioates, and may be a peptide nucleic acid. Polynucleotides may generally be assembled from isolated or cloned fragments of cDNA, genomic DNA, oligonucleotides, or individual nucleotides, or a combination thereof. Although the polynucleotides described herein are presented as DNA sequences, they include their corresponding RNA sequences and their complementary (e.g., fully complementary) DNA or RNA sequences, including their reverse complementary sequences.

Fragments of a polynucleotide can range from at least about 25 nucleotides, about 50 nucleotides, about 75 nucleotides, about 100 nucleotides, about 150 nucleotides, about 200 nucleotides, about 250 nucleotides, about 300 nucleotides, about 400 nucleotides, about 500 nucleotides, about 600 nucleotides, about 700 nucleotides, about 800 nucleotides, about 900 nucleotides, about 1000 nucleotides, about 1100 nucleotides, about 1200 nucleotides, about 1300 nucleotides, or about 1400 nucleotides, and up to a full-length polynucleotide encoding the polypeptides described herein.

A polynucleotide typically contains phosphodiester linkages, although in some cases polynucleotide analogs are included that may have alternative backbones containing, for example, phosphoramidate, phosphorothioate, phosphorodithioate, or O-methylphosphoramidite linkages; and peptide polynucleotide backbones and linkages. Other polynucleotide analogs include polynucleotides with positively charged backbones, nonionic backbones, and ribose-free backbones. Modifications to the ribose phosphate backbone can be made for a variety of reasons, such as to improve the stability and half-life of such molecules in physiological environments or as probes on a biochip. Mixtures of naturally occurring polynucleotides and analogs can be prepared; alternatively, mixtures of different polynucleotide analogs and mixtures of naturally occurring polynucleotides and analogs can be prepared.

Numerous polynucleotide analogs are known, including, for example, those with phosphoramidate, phosphorothioate, phosphorodithioate, or O-methylphosphoramidite linkages and peptide polynucleotide backbones and linkages. Other polynucleotide analogs include those with positively charged backbones, nonionic backbones, and ribose-free backbones. Polynucleotides containing one or more carbocyclic sugars are also included.

Other analogues include peptide polynucleotides, which are those based on peptide analogues of polynucleotides.

Among the uses of the disclosed polynucleotides and fragments thereof is the use of the fragments as probes in hybridization-based assays or as primers for use in amplification-based assays. Such fragments typically comprise at least about 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 or more contiguous nucleotides from a DNA sequence. In other embodiments, a DNA fragment comprises at least about 10, 15, 20, 30, 40, 50, or 60 or more contiguous nucleotides from a DNA sequence. Thus, according to one aspect, a method for detecting a polynucleotide is also provided, comprising using probes or primers, or both.

The main parameters influencing the choice of hybridization conditions and guidelines for developing suitable conditions are described in Sambrook, J., E. F. Fritsch, and T. Maniatis (1989, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.). Using the genetic code data in combination with the polypeptide sequences described herein, sets of degenerate oligonucleotides can be generated. Such oligonucleotides are useful as primers, for example, in polymerase chain reactions (PCR), which isolate and amplify DNA fragments.

At least one modification (e.g., mutation) may be included in one or more of NtINV3-S , NtINV3-T , NtINV4-S , and NtINV4-T .

At least one modification (eg, mutation) may be included in one or more of NtINV4-S and NtINV4-T .

At least one modification (e.g., mutation) may be included in one or more of NtINV3-S , NtINV3-T , NtINV4-S , and NtINV4-T . Optionally, at least one or more additional modifications (e.g., mutations) may be included in one or more of NtSUS1-S, NtSUS1-T, NtSUS2-S, NtSUS2-T, NtSUS3-S, NtSUS3-T, NtSUS4-S, NtSUS4-T, NtSUS5-S, NtSUS5-T, NtSUS6-S, and NtSUS6-T, preferably in one or more of NtSUS2-S, NtSUS2-T, NtSUS3-S, NtSUS3-T, NtSUS4-S, and NtSUS4-T .

At least one modification (e.g., mutation) can be included in one or more of NtINV3-S , NtINV3-T , NtINV4-S , and NtINV4-T , and at least one modification (e.g., mutation) can be included in one or more of NtSUS2-S, NtSUS2-T, NtSUS3-S, NtSUS3-T, NtSUS4-S, and NtSUS4-T , whereas the modification(s) (e.g., mutation(s)) is(are) not included in one or more of NtSUS1-S, NtSUS1-T, NtSUS5-S, NtSUS5-T, NtSUS6-S, and NtSUS6-T .

At least one modification (e.g., mutation) can be included in one or more of NtINV3-S , NtINV3-T , NtINV4-S , and NtINV4-T , and at least one modification (e.g., mutation) can be included in one or more of NtSUS2-S, NtSUS3-S, NtSUS3-T , and NtSUS4-S , whereas the modification(s) (e.g., mutation(s)) is(are) not included in one or more of NtSUS1-S, NtSUS1-T, NtSUS2-T, NtSUS4-T, NtSUS5-S, NtSUS5-T, NtSUS6-S, and NtSUS6-T .

At least one modification (e.g., mutation) may be included in one or more ofNtINV4-S And NtINV4-T, and by at least one modification (e.g. mutation) may be included in one or more of theNtSUS2-S, NtSUS2-T, NtSUS3-S, NtSUS3-T, NtSUS4-S And NtSUS4-T, whereas the modification(s) (e.g. mutation(s)) is(are) not included in one or more ofNtSUS1-S, NtSUS1-T, NtSUS5-S, NtSUS5-T, NtSUS6-SAndNtSUS6-T.

At least one modification (e.g., mutation) can be included in one or more of NtINV4-S and NtINV4-T , and at least one modification (e.g., mutation) can be included in one or more of NtSUS2-S, NtSUS3-S, NtSUS3-T and NtSUS4-S , whereas the modification(s) (e.g., mutation(s)) is(are) not included in one or more of NtSUS1-S, NtSUS1-T, NtSUS2-T, NtSUS4-T, NtSUS5-S, NtSUS5-T, NtSUS6-S and NtSUS6-T .

3. Polypeptide

Also provided is an isolated polypeptide comprising, consisting of, or consisting essentially of a polypeptide having at least 60% sequence identity to any of the polypeptides described herein, including any of the polypeptides listed in the sequence listing. Preferably, the isolated polypeptide comprises, consists of, or essentially consists of a sequence having at least 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 75%, 80%, 85%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, or 100% sequence identity thereto.

Also provided is an NtINV polypeptide comprising, consisting of, or consisting essentially of a sequence characterized by at least 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9% or 100% sequence identity to SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6 or SEQ ID NO: 8.

Also provided is a polypeptide comprising, consisting of, or essentially consisting of a sequence having at least 80%, 81%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, or 100% sequence identity to SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, or SEQ ID NO: 8.

Also provided is a polypeptide comprising, consisting of, or essentially consisting of a sequence having at least 95%, 96%, 97%, 98%, 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, or 100% sequence identity to SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, or SEQ ID NO: 8.

Also provided is a polypeptide encoded by SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, or SEQ ID NO: 8.

The polypeptide may provide sequences that have a sufficient or substantial degree of identity or similarity to SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, or SEQ ID NO: 8 to perform the function of INV.

Also provided is an NtSUS polypeptide comprising, consisting of, or consisting essentially of a sequence characterized by at least 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9% or 100% sequence identity to SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23, SEQ ID NO: 25, SEQ ID NO: 27, SEQ ID NO: 29 or SEQ ID NO: 31; or SEQ ID NO: 33.

Also provided is a polypeptide comprising, consisting, or consisting essentially of a sequence having at least 80%, 81%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, or 100% sequence identity to SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23, SEQ ID NO: 25, SEQ ID NO: 27, SEQ ID NO: 29 or SEQ ID NO: 31; or SEQ ID NO: 33.

Also provided is a polypeptide comprising, consisting or essentially consisting of a sequence having at least 95%, 96%, 97%, 98%, 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9% or 100% sequence identity to SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23, SEQ ID NO: 25, SEQ ID NO: 27, SEQ ID NO: 29 or SEQ ID NO: 31; or SEQ ID NO: 33.

Also provided is a polypeptide encoded by SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23, SEQ ID NO: 25, SEQ ID NO: 27, SEQ ID NO: 29, or SEQ ID NO: 31; or SEQ ID NO: 33.

The polypeptide may provide sequences having a sufficient or substantial degree of identity or similarity to SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23, SEQ ID NO: 25, SEQ ID NO: 27, SEQ ID NO: 29, or SEQ ID NO: 31, or SEQ ID NO: 33 to perform the SUS function.

Typically, fragments of the polypeptide(s) retain some or all of the function or activity of the full-length sequence, such as INV or SUS activity. Fragments of the polypeptide can range from at least about 25 amino acids, about 50 amino acids, about 75 amino acids, about 100 amino acids, about 150 amino acids, about 200 amino acids, about 250 amino acids, about 300 amino acids, about 400 amino acids, about 500 amino acids, and up to the full-length polypeptide described herein.

Polypeptides also include mutant variants obtained by introducing any type of adjustments (e.g., insertion, deletion, or substitution of amino acids; changes in glycosylation status; changes that affect refolding or isomerization, three-dimensional structures, or self-association states), which may be intentionally engineered or naturally isolated, provided that they still retain some or all of their functionality or activity. Preferably, this function or activity is modulated.

A deletion refers to the removal of one or more amino acids from a polypeptide. An insertion refers to one or more amino acid residues introduced into a predetermined location in a polypeptide. Insertions may involve intrasequence insertions of one or more amino acids. A substitution refers to the replacement of amino acids of a polypeptide with other amino acids having similar properties (such as similar hydrophobicity, hydrophilicity, antigenicity, propensity to form or break α-helical structures or β-sheet structures). Amino acid substitutions typically involve a single residue, but may be grouped depending on the functional constraints imposed on the polypeptide and may range from about 1 to about 10 amino acids. Amino acid substitutions are preferably conservative amino acid substitutions, as described below. Amino acid substitutions, deletions, or insertions can be accomplished using peptide synthesis techniques, such as solid phase peptide synthesis, or by manipulation of recombinant DNA. Methods for manipulating DNA sequences to produce substitution, insertion or deletion variants of a polypeptide are well known in the art. A variant may be characterized by the presence of adjustments that cause a "silent" change to occur and result in a functionally equivalent polypeptide. Intentional amino acid substitutions may be made based on similarity of residues with respect to polarity, charge, solubility, hydrophobicity, hydrophilicity and amphipathic nature, so long as the binding that determines the secondary structure of the substance is maintained. For example, negatively charged amino acids include aspartic acid and glutamic acid, positively charged amino acids include lysine and arginine, and amino acids with uncharged polar end groups characterized by similar hydrophilicity values include leucine, isoleucine, valine, glycine, alanine, asparagine, glutamine, serine, threonine, phenylalanine and tyrosine. Conservative substitutions can be made, for example, according to the table below. Amino acids in the same block in the second column and preferably in the same row in the third column can be substituted for one another:

ALIPHATIC Non-polar Gly Ala Pro Ile Leu Val Polar - uncharged Cys Ser Thr Met Asn Gly Polar - charged Asp Glu Lys Arg AROMATIC His Phe Trp Tyr

The polypeptide may be a mature polypeptide, or an immature polypeptide, or a polypeptide derived from an immature polypeptide. The polypeptides may be in linear form or cyclized using known methods. The polypeptides typically contain at least 10, at least 20, at least 30, or at least 40 contiguous amino acids.

At least one modification (eg, mutation) may be included in one or more of NtINV3-S, NtINV3-T, NtINV4-S, and NtINV4-T.

At least one modification (eg, mutation) may be included in one or more of NtINV4-S and NtINV4-T.

At least one modification (e.g., mutation) may be included in one or more of NtINV3-S, NtINV3-T, NtINV4-S, and NtINV4-T. Optionally, at least one or more additional modifications (e.g., mutations) may be included in one or more of NtSUS1-S, NtSUS1-T, NtSUS2-S, NtSUS2-T, NtSUS3-S, NtSUS3-T, NtSUS4-S, NtSUS4-T, NtSUS5-S, NtSUS5-T, NtSUS6-S, and NtSUS6-T, preferably in one or more of NtSUS2-S, NtSUS2-T, NtSUS3-S, NtSUS3-T, NtSUS4-S, and NtSUS4-T.

At least one modification (e.g., mutation) can be included in one or more of NtINV3-S, NtINV3-T, NtINV4-S, and NtINV4-T, and at least one modification (e.g., mutation) can be included in one or more of NtSUS2-S, NtSUS2-T, NtSUS3-S, NtSUS3-T, NtSUS4-S, and NtSUS4-T, whereas the modification(s) (e.g., mutation(s)) is(are) not included in one or more of NtSUS1-S, NtSUS1-T, NtSUS5-S, NtSUS5-T, NtSUS6-S, and NtSUS6-T.

At least one modification (e.g., mutation) can be included in one or more of NtINV3-S, NtINV3-T, NtINV4-S, and NtINV4-T, and at least one modification (e.g., mutation) can be included in one or more of NtSUS2-S, NtSUS3-S, NtSUS3-T, and NtSUS4-S, whereas the modification(s) (e.g., mutation(s)) is(are) not included in one or more of NtSUS1-S, NtSUS1-T, NtSUS2-T, NtSUS4-T, NtSUS5-S, NtSUS5-T, NtSUS6-S, and NtSUS6-T.

At least one modification (e.g., mutation) can be included in one or more of NtINV4-S and NtINV4-T, and at least one modification (e.g., mutation) can be included in one or more of NtSUS2-S, NtSUS2-T, NtSUS3-S, NtSUS3-T, NtSUS4-S and NtSUS4-T, whereas the modification(s) (e.g., mutation(s)) is(are) not included in one or more of NtSUS1-S, NtSUS1-T, NtSUS5-S, NtSUS5-T, NtSUS6-S and NtSUS6-T.

At least one modification (e.g., mutation) can be included in one or more of NtINV4-S and NtINV4-T, and at least one modification (e.g., mutation) can be included in one or more of NtSUS2-S, NtSUS3-S, NtSUS3-T and NtSUS4-S, whereas the modification(s) (e.g., mutation(s)) is(are) not included in one or more of NtSUS1-S, NtSUS1-T, NtSUS2-T, NtSUS4-T, NtSUS5-S, NtSUS5-T, NtSUS6-S and NtSUS6-T.

4. Plant modification

Transformation

Recombinant constructs can be used to transform plants or plant cells to modulate the expression, function, or activity of a polypeptide. A recombinant polynucleotide construct can comprise a polynucleotide encoding one or more polynucleotides described herein operably linked to a regulatory region suitable for expression of the polypeptide. Thus, a polynucleotide can comprise a coding sequence that encodes a polypeptide described herein. Plants or plant cells in which the expression, function, or activity of a polypeptide is modulated can include mutant, non-naturally occurring, transgenic, man-made, or genetically engineered plants or plant cells. Preferably, the transgenic plant or plant cell comprises a genome that has been corrected by the stable integration of recombinant DNA. Recombinant DNA comprises DNA obtained by genetic engineering techniques and constructed outside a cell, and comprises DNA comprising naturally occurring DNA, or cDNA, or synthetic DNA. A transgenic plant may include a plant regenerated from an initially transformed plant cell and transgenic plants that are progeny of later generations or hybrids of the transformed plant. Preferably, the transgenic modification provides for an adjustment in the expression or function or activity of the polynucleotide or polypeptide described herein compared to that of a control plant.

The polypeptide encoded by the recombinant polynucleotide may be a native polypeptide or may be heterologous to the cell. In some cases, the recombinant construct comprises a polynucleotide that provides expression modulation operably linked to a regulatory region. Examples of suitable regulatory regions are described herein.

Also provided are vectors containing recombinant polynucleotide constructs such as those described herein. Suitable vector bases include, for example, those commonly used in the art, such as plasmids, viruses, artificial chromosomes, bacterial artificial chromosomes, yeast artificial chromosomes, or bacteriophage artificial chromosomes. Suitable expression vectors include, but are not limited to, plasmids and viral vectors derived from, for example, bacteriophage, baculoviruses, and retroviruses. Numerous vectors and expression systems are commercially available.

Vectors may contain, for example, origins of replication, nuclear matrix binding sites, or markers. A marker gene may confer a selectable phenotype on a plant cell. For example, a marker may confer resistance to biocidal agents, such as resistance to an antibiotic (e.g., kanamycin, G418, bleomycin, or hygromycin) or to an herbicide (e.g., glyphosate, chlorsulfuron, or phosphinothricin). Additionally, the expression vector may contain a tag sequence designed to facilitate manipulation or detection (e.g., purification or localization) of the expressed polypeptide. Tag sequences, such as luciferase, beta-glucuronidase, green fluorescent polypeptide, glutathione S-transferase, polyhistidine, c-myc, or hemagglutinin sequences, are typically expressed as a fusion fragment to the encoded polypeptide. Such tags can be inserted anywhere within the polypeptide, including at the carboxyl or amino terminus.

A plant or plant cell can be transformed by integrating a recombinant polynucleotide into its genome to ensure its stable transformation. The plant or plant cell described herein can be stably transformed. Stably transformed cells typically retain the introduced polynucleotide with each cell division. The plant or plant cell can also be transiently transformed, such that the recombinant polynucleotide is not integrated into its genome. Transiently transformed cells typically lose all or some portion of the introduced recombinant polynucleotide with each cell division, such that the introduced recombinant polynucleotide cannot be detected in daughter cells after a sufficient number of cell divisions.

A number of techniques are available in the art for plant cell transformation, including biolistics; gene gun techniques; Agrobacterium-mediated transformation; viral vector-mediated transformation; freeze-thaw-based methods; microparticle bombardment; direct DNA uptake; ultrasonic treatment; microinjection; plant virus-mediated transfer; and electroporation.

If a cell or cultured tissue is used as a recipient tissue for transformation, plants can be regenerated from the transformed cultures, if desired, using methods known to those skilled in the art.

The selection of regulatory regions to be included in a recombinant construct depends on several factors, including but not limited to efficiency, selectability, inducibility, desired expression level, and preferential expression in certain cells or tissues. A routine task for one skilled in the art is to modulate the expression of a coding sequence by properly selecting regulatory regions and placing them relative to the coding sequence. Transcription of a polynucleotide can be modulated in a similar manner. Some suitable regulatory regions initiate transcription exclusively or preferentially in certain cell types. Methods for identifying and characterizing regulatory regions in plant genomic DNA are well known in the art.

Exemplary promoters include tissue-specific promoters recognized by tissue-specific factors present in different tissues or cell types (e.g., root-specific promoters, shoot-specific promoters, xylem-specific promoters), or present at different stages of development, or present in response to different environmental conditions. Suitable promoters include constitutive promoters that can be activated in most cell types without requiring specific inducers. Examples of promoters that can be used to control expression of a polypeptide include the cauliflower mosaic virus 35S (CaMV/35S) promoter, SSU, OCS, lib4, usp, STLS1, B33, nos, or ubiquitin or phaseolin gene promoters. Those skilled in the art can create many variations of recombinant promoters.

Tissue-specific promoters are transcriptional control elements that are active only in certain cells or tissues at certain times during plant development, such as vegetative tissues or reproductive tissues. Examples of tissue-specific promoters under developmental control include promoters that can initiate transcription only (or primarily only) in certain tissues, such as vegetative tissues, such as roots or leaves, or reproductive tissues such as fruits, ovules, seeds, pollen, stamens, flowers, or any embryonic tissue. Promoters specific to reproductive tissues may be, for example, anther-specific, ovule-specific, embryo-specific, endosperm-specific, integument-specific, seed and seed coat-specific, pollen-specific, petal-specific, sepal-specific, or combinations thereof.

Illustrative leaf-specific promoters include the pyruvate phosphate dikinase (PPDK) gene promoter from a C4 plant (maize), the cab-m1Ca+2 promoter from maize, the myb-related gene promoter from Arabidopsis thaliana (Atmyb5), ribulose bisphosphate carboxylase (RBCS) promoters (e.g., the tomato RBCS 1, RBCS2, and RBCS3A genes expressed in leaves and light-grown seedlings, RBCS1 and RBCS2 expressed in developing tomato fruit, or the ribulose bisphosphate carboxylase gene promoter expressed at high levels almost exclusively in mesophyll cells of leaf blades and leaf axils).

Exemplary senescent tissue-specific promoters include a tomato promoter active during fruit ripening, senescence and leaf abscission, a promoter of the gene encoding maize cysteine protease, the 82E4 promoter and a promoter of the SAG genes. Exemplary anther-specific promoters can be used. Exemplary promoters for preferential expression in the root, known to those skilled in the art, can be selected. Exemplary promoters for preferential expression in seeds include both seed-specific promoters (promoters active during seed development, such as seed storage polypeptide promoters) and germinating seed promoters (promoters active during seed germination).

Examples of inducible promoters include those responsive to pathogens, anaerobic conditions, elevated temperature, light, drought, low temperature, or high salt concentrations. Pathogen-inducible promoters include promoters of pathogenesis-related polypeptides (PR polypeptides), which are induced following pathogen infection (e.g., PR polypeptides, SAR polypeptides, beta-1,3-glucanase, chitinase).

In addition to plant promoters, other suitable promoters may be of bacterial origin, such as the octopine synthase gene promoter , the nopaline synthase gene promoter, and other Ti plasmid-derived promoters, or they may be derived from viral promoters (e.g., the 35S and 19S RNA promoters of cauliflower mosaic virus (CaMV), the constitutive promoters of tobacco mosaic virus, the 19S and 35S promoters of cauliflower mosaic virus (CaMV), or the 35S promoter of figwort mosaic virus).

Mutation

Disclosed is a plant or plant cell comprising at least one mutation in one or more polynucleotides or polypeptides described herein, wherein said mutation results in modulation of the function or activity of NtINV or the polypeptide(s) encoded therefor, or modulation of the function or activity of NtINV and NtSUS or the polypeptides encoded therefor. Combinations of such mutations are discussed herein.

A method is provided for modulating the level of an NtINV polypeptide or an NtINV polypeptide and an NtSUS polypeptide in a (dried) plant or in (dried) plant material, wherein said method comprises introducing into the genome of said plant one or more mutations that provide for modulation of the expression of at least one NtINV gene or at least one NtINV gene and at least one NtSUS gene, wherein said at least one gene is selected from any sequence according to the present invention.

Also provided is a method for identifying a plant with modulated levels of reducing sugars, said method comprising screening a polynucleotide sample from a plant of interest for the presence of one or more mutations in sequences according to the present disclosure, such as NtINV or NtINV and NtSUS or a combination thereof, and optionally comparing the identified mutation(s) with mutation(s) known to provide modulation of reducing sugar levels.

Also disclosed is a plant or plant cell that is heterozygous or homozygous for one or more mutations in the NtINV gene or the NtINV gene and the NtSUS gene according to the present invention, wherein said mutation results in modulated expression of the gene, or function, or activity of the NtINV polypeptide or the NtINV and NtSUS polypeptides encoded by them.

A variety of approaches, including sexual crossing, can be used to combine mutations in a single plant. A plant characterized by the presence of one or more favorable mutations in a heterozygous or homozygous state in a gene according to the present invention that modulate the expression of the gene or the function or activity of the polypeptide encoded by it can be crossed with a plant characterized by the presence of one or more favorable mutations in a heterozygous or homozygous state in one or more genes that modulate their expression or the function or activity of the polypeptide encoded by them. In one embodiment, the crosses are carried out to introduce one or more favorable mutations in a heterozygous or homozygous state into a gene in accordance with the present invention in the same plant.

The function or activity of one or more polypeptides of the present invention in a plant is increased or decreased if the function or activity is lower or higher than the function or activity of the same polypeptide(s) in a plant that has not been modified to suppress the function or activity of that polypeptide and that has been cultured, harvested, and dried using the same protocols.

In some embodiments, the mutation(s) are introduced into the plant or plant cell using a mutagenesis approach, and the introduced mutation is identified or selected for using methods known to those skilled in the art, such as Southern blot analysis, DNA sequencing, PCR analysis, or phenotypic analysis. Mutations that affect gene expression or that disrupt the function of the encoded polypeptide can be identified using methods well known in the art. Insertional mutations in exons of a gene typically result in non-functional mutant variants. Mutations in conserved residues can be particularly effective in suppressing the metabolic function of the encoded polypeptide. For example, it is understood that a mutation in one or more highly conserved regions will likely provide an adjustment in the function of the polypeptide, while a mutation outside of these highly conserved regions will likely have little or no effect on the function of the polypeptide. Additionally, a mutation in a single nucleotide can result in the creation of a stop codon, the presence of which will result in a truncated polypeptide and, depending on the degree of truncation, loss of function.

Methods for producing mutant polynucleotides and polypeptides are also disclosed. Any plant of interest, including a plant cell or plant material, can be genetically modified by a variety of methods known to induce mutagenesis, including site-directed mutagenesis, oligonucleotide-directed mutagenesis, chemically induced mutagenesis, ionizing radiation-induced mutagenesis, modified base mutagenesis, gapped DNA duplex mutagenesis, double-strand break mutagenesis, reparation-deficient host strain mutagenesis, whole gene synthesis mutagenesis, DNA shuffling, and other equivalent methods.

Mutations in the polynucleotides and polypeptides described herein may include mutations introduced by man, or artificial mutations, or mutations created by genetic engineering. Mutations in the polynucleotides and polypeptides described herein may be mutations that are obtained or that can be obtained by a method that includes a manipulation step in vitro or in vivo . Mutations in the polynucleotides and polypeptides described herein may be mutations that are obtained or that can be obtained by a method that involves human intervention. The function or activity of a mutant version of a polypeptide may be higher, lower, or approximately the same as that of a polypeptide that has not been subjected to mutation.

Methods by which a mutation is introduced randomly into a polynucleotide may include chemical mutagenesis and radiation mutagenesis. Chemical mutagenesis involves the use of exogenously added chemicals, such as mutagenic, teratogenic, or carcinogenic organic compounds, to induce mutations. Mutagens that create primarily point mutations and short deletions, insertions, missense mutations, simple repeats, transversions, or transitions, including chemical mutagens or radiation, may be used to create mutations. Mutagens include ethyl methanesulfonate, methyl methanesulfonate, N-ethyl-N-nitrosourea, triethylmelamine, N-methyl-N-nitrosourea, procarbazine, chlorambucil, cyclophosphamide, diethyl sulfate, acrylamide monomer, melphalan, nitrogen mustard, vincristine, dimethylnitrosamine, N-methyl-N'-nitro-nitrosoguanidine, nitrosoguanidine, 2-aminopurine, 7,12-dimethylbenz(a)anthracene, ethylene oxide, hexamethylphosphoramide, bisulfan, diepoxyalkanes (diepoxyoctane, diepoxybutane, etc.), 2-methoxy-6-chloro-9[3-(ethyl-2-chloroethyl)aminopropylamino]acridine dihydrochloride and formaldehyde.

Spontaneous mutations at the locus are also contemplated, which may not be directly caused by the mutagen, provided that they result in the desired phenotype. Suitable mutagenic agents may also include, for example, ionizing radiation, such as X-rays, gamma rays, fast neutron radiation, and ultraviolet radiation. The dose of the mutagenic chemical or radiation is determined experimentally for each type of plant tissue so as to obtain a mutation rate that is below the threshold level characterized by lethality or reproductive sterility. Any method for producing a plant polynucleotide known to those skilled in the art can be used to produce a plant polynucleotide for screening for mutations.

The mutation process may involve one or more stages of plant crossing.

After introducing a mutation, screening can be performed to identify mutations that result in genes with premature stop codons or otherwise non-functional genes. After introducing a mutation, screening can be performed to identify mutations that result in functional genes that are capable of being expressed at increased or decreased levels. Screening for mutant variants can be performed by sequencing or by using one or more probes or primers specific for a given gene or polypeptide. Polynucleotides can also be designed to have specific mutations that can result in modulation of gene expression, modulation of mRNA stability, or modulation of polypeptide stability. Such plants are referred to herein as "non-naturally occurring" or "mutant" plants. Typically, mutant or non-naturally occurring plants contain at least a portion of a foreign or synthetic or man-made nucleotide (e.g., DNA or RNA) that was not present in the plant prior to manipulation. The foreign nucleotide may be one nucleotide, two or more nucleotides, two or more contiguous nucleotides, or two or more non-contiguous nucleotides, such as at least 10, 20, 30, 40, 50, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000, 1100, 1200, 1300, 1400, or 1500 or more contiguous or non-contiguous nucleotides.

Transgenics and genome editing

In order to modulate the expression or function or activity of one or more polynucleotides or polypeptides described herein, one can use sequence-specific polynucleotides that can disrupt the transcription of one or more endogenous genes; sequence-specific polynucleotides that can disrupt the translation of RNA transcripts (e.g., double-stranded RNA, siRNA, ribozymes); sequence-specific polynucleotides that can disrupt the stability of one or more polypeptides; sequence-specific polynucleotides that can disrupt the enzymatic function of one or more polypeptides or the binding function of one or more polypeptides with respect to substrates or regulatory polypeptides; antibodies that exhibit specificity for one or more polypeptides; small molecules that can disrupt the stability of one or more polypeptides or the enzymatic function of one or more polypeptides or the binding function of one or more polypeptides; zinc finger polypeptides that bind one or more polynucleotides; and meganucleases that function with respect to one or more polynucleotides. Genome editing technologies are well known in the art and are discussed further below.

Zinc finger nucleases

Zinc finger polypeptides can be used to modulate the expression or function or activity of one or more of the NtINV or NtINV and NtSUS polynucleotides described herein. The use of zinc finger nucleases is described in Nature Rev. Genet. (2010) 11(9):636-646).

e. Meganucleases

Meganucleases, such as I-CreI, can be used to modulate the expression or function or activity of one or more of the NtINV or NtINV and NtSUS polynucleotides described herein. The use of meganucleases is described in Curr Gene Ther. (2011) Feb;11(1):11-27 and Int J Mol Sci. (2019) 20(16), 4045.

f. TALEN

Transcription activator-like effector nucleases (TALENs) can be used to modulate the expression or function or activity of one or more of the NtINV or NtINV and NtSUS polynucleotides described herein. The use of TALENs is described in Nature Rev. Mol. Cell Biol . (2013) 14: 49–55 and Int J Mol Sci. (2019) 20(16), 4045.

g. CRISPR

The CRISPR system can be used to modulate the expression or function or activity of one or more of the NtINV or NtINV and NtSUS polynucleotides described herein, and is a preferred method. This technology is described, for example, in Plant Methods (2016) 12:8; Front Plant Sci. (2016) 7: 506; Biotechnology Advances (2015) 33, 1, p41-52; Acta Pharmaceutica Sinica B (2017) 7, 3, p292-302; Curr. Op. in Plant Biol. (2017) 36, 1-8 and Int J Mol Sci (2019) 20(16), 4045. As is well known in the art, a CRISPR editing system typically comprises two components: a CRISPR-associated endonuclease (Cas) (e.g., Cas9) and a guide RNA (gRNA). Cas forms a double-stranded DNA break at a site in the genome that is determined by the sequence of the gRNA molecule bound to Cas. The site of the DNA break by Cas is determined by the unique sequence of the gRNA bound to it. The gRNA is a specially designed RNA sequence that recognizes a site of interest in the target DNA and directs the Cas nuclease there for editing. It consists of two parts: (i) tracrRNA, which serves as a scaffold for the binding of the Cas nuclease; and (ii) crisprRNA (crRNA) - a 17-20 nucleotide sequence complementary to the target DNA. The exact region of DNA to be targeted will depend on the particular application. For example, to provide activation or repression of a target polynucleotide, a gRNA can be targeted to a promoter that drives expression of the target polynucleotide. Methods for constructing gRNA are well known in the art, including methods using Harvard's Chop Chop.

Applications of Cas9-based genome editing in Arabidopsis and tobacco are described, for example, in Methods Enzymol. (2014) 546:459–72 and Plant Physiol Biochem . (2018) 131:37–46. CRISPR technology has been widely applied in plants (see, for example, WO 2015/189693).

In addition to Cas9, other RNA-guided nucleases have been described for use in the CRISPR system, including: Casl, CaslB, Cas2, Cas3, Cas4, Cas5, Cas6, Cas7, Cas8, CaslO, Cpfl, Csyl, Csy2, Csy3, Csel, Cse2, Cscl, Csc2, Csa5, Csn2, Csm2, Csm3, Csm4, Csm5, Csm6, Cmrl, Cmr3, Cmr4, Cmr5, Cmr6, Csbl, Csb2, Csb3, Csxl7, Csxl4, CsxlO, Csxl6, CsaX, Csx3, Csxl, Csxl5, Csfl, Csf2, Csf3 and Csf4. In certain embodiments, use of Cas9 is preferred.

The present disclosure further provides a CRISPR-based genome editing system comprising an RNA-guided nuclease and a gRNA, wherein the CRISPR-based genome editing system modulates the activity of one or more polynucleotides described herein. The present invention also provides a method of cleaving one or more polynucleotides in a plant cell, comprising introducing a gRNA and an RNA-guided nuclease into the plant cell, wherein the gRNA acts in combination with the RNA-guided nuclease to create a strand break in one or more polynucleotides described herein. Also disclosed is a CRISPR construct comprising: (i) a polynucleotide encoding a CRISPR-associated endonuclease; and (ii) a gRNA comprising a polynucleotide sequence (typically about 17-20 nucleotides) complementary to the DNA of a polynucleotide described herein that is to be targeted.

h. Antisense modification

Antisense oligonucleotide technology is another well-known method that can be used to modulate the expression or activity of one or more NtINV polypeptides or one or more NtINV and NtSUS polypeptides. See, e.g., Gene (1988) 10;72(1-2):45-50.

i. Mobile genetic elements

Alternatively, targeting of genes for inactivation can be accomplished by introducing transposons (e.g., IS elements) into the genomes of plants of interest. See, e.g., Cytology and Genetics (2006) 40(4):68–81.

j. Ribozymes

Alternatively , NtINV or NtINV and NtSUS polynucleotides can be targeted for inactivation by introducing ribozymes derived from a number of small circular RNAs that are capable of self-cleavage and replication in plants. See, for example, FEMS Microbiology Reviews (1999) 23, 3, 257-275.

5. Plants

Mutant or non-naturally occurring plants or plant cells may contain any combination of one or more modifications (e.g., mutations) in one or more of NtINV or NtINV and NtSUS , one or more of NtINV or NtINV and NtSUS that result in modulation of the expression or function or activity of these polynucleotides or products of such polynucleotides. For example, mutant or non-naturally occurring plants or plant cells may have one modification in one NtINV or in one NtINV and one NtSUS polynucleotide or polypeptide; multiple modifications in one NtINV or one NtINV and one NtSUS polynucleotide or polypeptide; one modification in two or more, or three or more, or four or more NtINV or NtINV and NtSUS polynucleotides or polypeptides; or multiple modifications in two or more or three or more or four or more NtINV or NtINV and NtSUS polynucleotides or polypeptides. As a further example, mutant or non-naturally occurring plants or plant cells may comprise one or more modifications in a specific portion of the NtINV or NtINV and NtSUS polynucleotide(s) or polypeptide(s), such as in a region of NtINV or NtINV and NtSUS that encodes the active site of the NtINV or NtSUS polypeptide or a portion thereof. As a further example, mutant or non-naturally occurring plants or plant cells may comprise one or more modifications in a region outside of one or more NtINV or NtINV and NtSUS polynucleotide(s) or polypeptide(s), such as in a region upstream or downstream of the NtINV or NtINV and NtSUS polynucleotide(s) it regulates, so long as they modulate the function or expression of the NtINV or NtINV and NtSUS polypeptide(s). Upstream elements may include promoters, enhancers, or transcription factors. Some elements, such as enhancers, may be upstream or downstream of the gene they regulate. The element(s) are not necessarily located adjacent to the gene they regulate, as some elements have been found to be located several thousand base pairs upstream or downstream of the gene they regulate. Mutant or non-naturally occurring plants or plant cells may have one or more modifications located within the first 100 nucleotides of the gene(s), within the first 200 nucleotides of the gene(s), within the first 300 nucleotides of the gene(s), within the first 400 nucleotides of the gene(s), within the first 500 nucleotides of the gene(s), within the first 600 nucleotides of the gene(s), within the first 700 nucleotides of the gene(s), within the first 800 nucleotides of the gene(s), within the first 900 nucleotides of the gene(s), within the first 1000 nucleotides of the gene(s), within the first 1100 nucleotides of the gene(s), within the first 1200 nucleotides of the gene(s), within the first 1300 nucleotides of gene(s), within the first 1400 nucleotides of the gene(s), or within the first 1500 nucleotides of the gene(s). Mutant or non-naturally occurring plants or plant cells may have one or more modifications located within the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth, thirteenth, fourteenth, or fifteenth set of 100 nucleotides of the gene(s), or a combination thereof. Disclosed are mutant or non-naturally occurring plants or plant cells (e.g., mutant, non-naturally occurring, or transgenic plants or plant cells, etc., as described herein) comprising variants of a mutant polypeptide.

In one embodiment, plant seeds are mutagenized and then grown into first generation mutant plants. The first generation plants are then allowed to self-pollinate and second generation plants are grown from the seeds of the first generation plants and then tested for mutations at their loci. Although the mutated plant material can be screened for mutations, an advantage of screening second generation plants is that all somatic mutations correspond to germline mutations. One skilled in the art will appreciate that various plant material, including but not limited to seeds, pollen, plant tissue, or plant cells, can be mutagenized to create mutant plants. However, the type of plant material mutagenized can be important when a plant polynucleotide is screened for mutations. For example, if pollen is mutagenized prior to pollination of an unmutagenesis plant, first generation plants are grown from the seeds obtained from that pollination. Every cell in the first generation of plants will contain the mutations that arose in the pollen; thus, these first generation plants can then be screened for mutations instead of waiting for the second generation to emerge.

6. Obtaining modified plants, screening and crossing

NtINV or NtINV and NtSUS polynucleotides obtained from individual plants, plant cells or plant material may optionally be pooled to expedite screening for mutations in a population of plants derived from mutagenized plant tissue, plant cells or plant material. One or more subsequent generations of plants, plant cells or plant material may be screened. The size of the optionally pooled group depends on the sensitivity of the screening method used.

After the samples are optionally pooled, they can be analyzed using polynucleotide-specific amplification techniques such as PCR. Any one or more primers or probes specific for a gene or sequences immediately adjacent to a gene can be used to amplify sequences within the optionally pooled sample. Preferably, one or more primers or probes are designed to amplify regions of the locus where useful mutations are most likely to occur. Most preferably, the primer is designed to detect mutations in regions of the polynucleotide. Additionally, it would be preferable for the primer(s) and probe(s) to avoid known polymorphic sites to facilitate screening for point mutations. To facilitate detection of the amplification products, one or more primers or probes can be labeled using any conventional labeling method. Primer(s) or probe(s) can be designed based on the sequences described herein using methods that are well known in the art.

To facilitate detection of the amplification products, the primer(s) or probe(s) may be labeled using any conventional labeling method. They may be designed based on the sequences described herein using methods that are well known in the art.

Polymorphisms can be identified using tools known in the art and some described in the literature.

In some embodiments, a plant can be regenerated or grown from a plant, plant tissue, or plant cell. Any suitable method for regenerating or growing a plant from a plant cell or plant tissue can be used, such as, but not limited to, tissue culture or protoplast regeneration. Preferably, plants can be regenerated by growing transformed plant cells on callus induction medium, shoot induction medium, or root induction medium. See, for example, McCormick et al., Plant Cell Reports 5:81-84 (1986). These plants can then be grown and either pollinated with the same transformed line or different lines and the resulting hybrid identified as having expression of the desired phenotypic characteristic. Two or more generations can be grown to ensure that expression of the desired phenotypic characteristic is stably maintained and heritable, and then the seeds collected to ensure that expression of the desired phenotypic characteristic has been achieved. Thus, the term "transformed seeds" refers to seeds that contain a nucleotide construct stably integrated into the plant genome.

Accordingly, in a further aspect, a method for producing a mutant plant is provided. The method comprises obtaining at least one plant cell comprising a NtINV or NtINV and NtSUS gene encoding a functional polynucleotide as described herein (or any combination thereof as described herein). The at least one plant cell is then treated under conditions effective to modulate the function of the polynucleotide(s). The at least one mutant plant cell is then propagated to produce a mutant plant, wherein the mutant plant has modulated levels of the NtINV or NtINV and NtSUS polypeptide(s) as described herein (or any combination thereof as described herein) as compared to those of control plants. In one embodiment of the method for producing a mutant plant, the treating step comprises exposing the at least one cell to a chemical mutagenic agent as described above and under conditions effective to produce at least one mutant plant cell. In another embodiment of the method, the treating step comprises exposing at least one cell to a source of ionizing radiation under conditions effective to produce at least one mutant plant cell. The term "mutant plant" includes mutant plants in which the genotype is modified compared to the genotype of a control plant, preferably by methods other than genetic engineering and genetic modification.

In certain embodiments, the mutant plant, mutant plant cell, or mutant plant material may comprise one or more mutations that occur naturally in another plant, plant cell, or plant material and provide the desired trait. The mutation may be introduced (e.g., by introgression) into another plant, plant cell, or plant material (e.g., a plant, plant cell, or plant material with a genetic background different from that of the plant from which the mutation originates) to provide the trait therein. Thus, by way of example, a mutation that occurs naturally in a first plant, can be introduced into a second plant, such as a second plant with a genetic background that differs from that of the first plant. In this way, one skilled in the art can search for and identify a plant that naturally carries in its genome one or more mutant alleles of the genes described herein that provide the desired trait. The mutant allele(s) that occur in nature can be transferred to the second plant by various methods, including selection, backcrossing, and introgression to produce lines, varieties, or hybrids that have one or more mutations in the genes described herein. The same technique can also be used to introgresse one or more non-naturally occurring mutation(s) from the first plant into the second plant. Plants that exhibit the desired trait can be selected from a pool of mutant plants. Preferably, the selection is performed using the polynucleotide data described herein. Accordingly, selection can be performed for a genetic trait compared to a control. Such a screening approach may involve the use of conventional amplification or hybridization techniques as discussed herein. Thus, a further aspect of the present invention relates to a method for identifying a mutant plant comprising the steps of (a) obtaining a sample containing one or more polynucleotides fromNtINV or NtINV And NtSUSfrom a plant; and (b) determining the sequence of the polynucleotide(s), wherein a difference in the sequence of the polynucleotide(s) compared to the polynucleotide(s) of a control plant indicates that said plant is a mutant plant. In another aspect, there is provided a method of identifying a mutant plant that accumulates increased or decreased levels of reducing sugar(s) compared to a control plant, comprising the steps of (a) obtaining a sample from a plant to be screened; (b) determining whether said sample contains one or more mutations in one or more polynucleotidesNtINV or NtINV And NtSUS, described herein; and (c) determining the level of at least one reducing sugar in said plant, preferably glucose or fructose or a combination thereof. Preferably, the level of at least one reducing sugar is determined in dried leaves. In another aspect, there is provided a method for producing a mutant plant that has increased or decreased levels of at least one reducing sugar, preferably glucose or fructose or a combination thereof, compared to a control plant, comprising the steps of (a) obtaining a sample from a first plant; (b) determining whether said sample contains one or more mutations in one or more polynucleotidesNtINV or NtINV And NtSUS, described herein, which result in modulated levels of at least one reducing sugar; and (c) transferring one or more mutations to a second plant. Preferably, the level of at least one reducing sugar is determined in dried leaves. The mutation(s) can be transferred to the second plant by various methods that are known in the art, such as genetic engineering, gene manipulation, introgression, plant breeding, backcrossing, etc. In one embodiment, the first plant is a naturally occurring plant. In one embodiment, the second plant has a genetic background that is different from that of the first plant. In another aspect, there is provided a method of producing a mutant plant that has increased or decreased levels of at least one reducing sugar, preferably glucose or fructose or a combination thereof, compared to a control plant, comprising the steps of (a) obtaining a sample from the first plant; (b) determining whether the said sample contains one or more mutations in one or more of the polynucleotidesNtINV or NtINV And NtSUS, described herein, which result in modulated levels of at least one reducing sugar; and (c) introgression of one or more mutations into a second plant. Preferably, the level of at least one reducing sugar is determined in dried leaves. In one embodiment, the introgression step comprises plant selection, optionally including backcrossing, etc. In one embodiment, the first plant is a naturally occurring plant. In one embodiment, the second plant has a genetic background that is different from that of the first plant. In one embodiment, the first plant is not a cultivar or elite cultivar. In one embodiment, the second plant is a cultivar or elite cultivar. A further aspect relates to a mutant plant (including a mutant plant that is a cultivar or elite cultivar) obtained or obtainable by the methods described herein. In certain embodiments, a mutant plant may contain one or more mutations localized only to a specific region of the plant, such as within the sequence of one or more polynucleotides.NtINV or NtINV And NtSUS, described herein. According to this embodiment, the remaining genomic sequence of the mutant plant will be the same or substantially the same as that of the plant prior to mutagenesis.

In certain embodiments, the mutant plants may comprise one or more mutations located in more than one region of the plant's genome, such as within the sequence of one or more of the NtINV or NtINV and NtSUS polynucleotides described herein and in one or more additional regions of the genome. In this embodiment, the remainder of the genomic sequence of the mutant plant will not be the same or substantially the same as that of the plant prior to mutagenesis. In certain embodiments, the mutant plants may lack one or more mutations in one or more, two or more, three or more, four or more, or five or more exons of the NtINV or NtINV and NtSUS polynucleotide(s) described herein; or may lack one or more mutations in one or more, two or more, three or more, four or more, or five or more introns of the NtINV or NtINV and NtSUS polynucleotide(s) described herein; or may lack one or more mutations in the promoter of the NtINV or NtINV and NtSUS polynucleotide(s) described herein; or may lack one or more mutations in the 3'-untranslated region of the NtINV or NtINV and NtSUS polynucleotide(s) described herein; or may lack one or more mutations in the 5'-untranslated region of the NtINV or NtINV and NtSUS polynucleotide(s) described herein; or may lack one or more mutations in the coding region of the NtINV or NtINV and NtSUS polynucleotide(s) described herein; or may lack one or more mutations in the non-coding region of the NtINV or NtINV and NtSUS polynucleotide(s) described herein; or any combination of two or more, three or more, four or more, five or more, or six or more portions thereof.

In a further aspect, there is provided a method for identifying a plant, plant cell or plant material comprising a mutation in a gene encoding an NtINV or NtINV and NtSUS polynucleotide described herein, comprising (a) subjecting a plant, plant cell or plant material to mutagenesis; (b) obtaining a sample from said plant, plant cell or plant material or descendants thereof; and (c) determining the polynucleotide sequence of the NtINV or NtINV and NtSUS gene(s), or a variant or fragment thereof, wherein a difference in said sequence is indicative of the presence of one or more mutations therein. This method also enables the selection of plants having mutation(s) that occur(s) in genomic regions that affect the expression of the NtINV or NtINV and NtSUS gene in the plant cell, such as the transcription initiation site, start codon, intron region, exon-intron boundary, terminator or stop codon.

7. Plant families, species, varieties, seeds and tissue culture

Plants suitable for use in the present invention include monocotyledonous and dicotyledonous plants and plant cell systems and may include members of the genera Camellia, Cannabis or Nicotiana. Suitable species of Camellia and Cannabis include Camellia sinensis (tea), Cannabis sativa , Cannabis indica and Cannabis ruderalis .

Various embodiments relate to mutant tobacco, non-naturally occurring tobacco, or transgenic tobacco plants or tobacco plant cells and can be applied to any species of the genus Nicotiana , including N. rustica and N. tabacum (e.g., LA B21, LN KY171, TI 1406, Basma, Galpao, Perique, Beinhart 1000-1, and Petico). Other species include N. acaulis, N. acuminata, N. africana, N. alata, N. ameghinoi, N. amplexicaulis, N. arentsii, N. attenuata, N. azambujae, N. benavidesii, N. benthamiana, N. bigelovii, N. bonariensis, N. cavicola, N. clevelandii, N. cordifolia, N. corymbosa, N. debneyi, N. excelsior, N. forgetiana, N. fragrans, N. glauca, N. glutinosa, N. goodspeedii, N. gossei, N. hybrida, N. ingulba, N. kawakamii, N. knightiana, N. langsdorffii, N. linearis, N. longiflora, N. maritima, N. megalosiphon, N. miersii, N. noctiflora, N. nudicaulis, N. obtusifolia, N. occidentalis, N. occidentalis subsp. hesperis, N. otophora, N. paniculata, N. pauciflora, N. petunioides, N. plumbaginifolia, N. quadrivalvis, N. raimondii, N. repanda, N. rosulata, N. rosulata subsp. ingulba, N. rotundifolia, N. setchellii, N. simulans, N. solanifolia, N. spegazzinii, N. stocktonii, N. suaveolens, N. sylvestris, N. thyrsiflora, N. tomentosa, N. tomentosiformis, N. trigonophylla, N. umbratica, N. undulata, N. velutina, N. wigandioides, and N. × sanderae. In one embodiment, the plant is N. tabacum.

The use of tobacco varieties and tobacco cultivars is also provided for in this document. A transgenic, non-naturally occurring or mutant plant may therefore be a tobacco variety or tobacco cultivar that contains one or more transgenes or one or more genetic mutations or a combination thereof. The genetic mutation(s) (e.g., one or more polymorphisms) may be mutations that do not naturally occur in a particular tobacco variety or tobacco cultivar (e.g., tobacco cultivar) or may be genetic mutation(s) that do occur in nature, provided that the mutation does not naturally occur in a particular tobacco variety or tobacco cultivar (e.g., tobacco cultivar).

Specifically, applicable varieties of Nicotiana tabacum include Burley type tobacco, dark type tobacco, flue-cured tobacco and oriental type tobacco. Non-limiting examples of varieties or cultivars are: BD 64, CC 101, CC 200, CC 27, CC 301, CC 400, CC 500, CC 600, CC 700, CC 800, CC 900, Coker 176, Coker 319, Coker 371 Gold, Coker 48, CD 263, DF911, DT 538 LC Galpao tobacco, GL 26H, GL 350, GL 600, GL 737, GL 939, GL 973, HB 04P, HB 04P LC, HB3307PLC, Hybrid 403LC, Hybrid 404LC, Hybrid 501 LC, K 149, K 326, K 346, K 358, K394, K 399, K 730, KDH 959, KT 200, KT204LC, KY10, KY14, KY 160, KY 17, KY 171, KY 907, KY907LC, KY14xL8 LC, Little Crittenden, McNair 373, McNair 944, msKY 14xL8, Narrow Leaf Madole, Narrow Leaf Madole LC, NBH 98, N-126, N-777LC, N-7371LC, NC 100, NC 102, NC 2000, NC 291, NC 297, NC 299, NC 3, NC 4, NC 5, NC 6, NC7, NC 606, NC 71, NC 72, NC 810, NC BH 129, NC 2002, Neal Smith Madole, OXFORD 207, PD 7302 LC, PD 7309 LC, PD 7312 LC, Perique tobacco, PVH03, PVH09, PVH19, PVH50, PVH51, R 610, R 630, R 7-11, R 7-12, RG 17, RG 81, RG H51, RGH 4, RGH 51, RS 1410, Speight 168, Speight 172, Speight 179, Speight 210, Speight 220, Speight 225, Speight 227, Speight 234, Speight G-28, Speight G-70, Speight H-6, Speight H20, Speight NF3, TI 1406, TI 1269, TN 86, TN86LC, TN 90, TN 97, TN97LC, TN D94, TN D950, TR (Tom Rosson) Madole, VA 309, VA359, AA 37-1, B13P, Xanthi (Mitchell-Mor), Bel-W3, 79-615, Samsun Holmes NN, KTRDC number 2 hybrid 49, Burley 21, KY8959, KY9, MD 609, PG01, PG04, PO1, PO2, PO3, RG11, RG 8, VA509, AS44, Banket A1, Basma Drama B84/31, Basma I Zichna ZP4/B, Basma Xanthi BX 2A, Batek, Besuki Jember, C104, Coker 347, Criollo Misionero, Delcrest, Djebel 81, DVH 405, Galpão Comum, HB04P, Hicks Broadleaf, Kabakulak Elassona, Kutsage E1, LA BU 21, NC 2326, NC 297, PVH 2110, Red Russian, Samsun, Saplak, Simmaba, Talgar 28, Wislica, Yayaldag, Prilep HC-72, Prilep P23, Prilep PB 156/1, Prilep P12-2/1, Yaka JK-48, Yaka JB 125/3, TI-1068, KDH-960, TI-1070, TW136, Basma, TKF 4028, L8, TKF 2002, GR141, Basma xanthi, GR149, GR153, Petit Havana. Sub-varieties of the above with low conversion of nicotine to nornicotine are also provided, even if they are not specifically listed in this document.

Embodiments also relate to compositions and methods for producing mutant plants, non-naturally occurring plants, hybrid plants, or transgenic plants that have been modified to modulate the expression or function of one or more of the NtINV or NtINV and NtSUS polynucleotide(s) described herein (or any combination thereof described herein). Advantageously, the resulting mutant plants, non-naturally occurring plants, hybrid plants, or transgenic plants can be similar in general appearance to, or substantially the same as, control plants. Various phenotypic characteristics, such as maturity, number of leaves per plant, stem height, leaf ingrowth angle, leaf size (width and length), internode distance, and blade-to-vein ratio can be assessed by field observation.

One aspect relates to a seed of a mutant plant, a non-naturally occurring plant, a hybrid plant or a transgenic plant described herein. Preferably, the seed is a tobacco seed. A further aspect relates to pollen or an ovule of a mutant plant, a non-naturally occurring plant, a hybrid plant or a transgenic plant described herein. Furthermore, there is provided a mutant plant, a non-naturally occurring plant, a hybrid plant or a transgenic plant described herein that further comprises a polynucleotide that provides for the development of male sterility.

Also provided is a tissue culture of regenerated cells of a mutant plant, a non-naturally occurring plant, a hybrid plant, or a transgenic plant or part thereof, as described herein, wherein the culture regenerates plants capable of expressing all the morphological and physiological characteristics of the parent. The regenerated cells include cells from leaves, pollen, embryos, cotyledons, hypocotyls, roots, root tips, anthers, flowers and parts thereof, ovules, shoots, stems, petioles, pith, and seed pods or callus or protoplasts derived therefrom.

The plant material described herein may be a cured tobacco material, such as a cured Virginia or Oriental tobacco material. The cured tobacco material may be a flue-cured, sun-cured, or air-cured tobacco material.

The CORESTA recommendation for tobacco curing is described in CORESTA Handbook No. 17, April 2016, Sustainability in Leaf Tobacco Production.

8. Modulation of sugar content

The mutant, transgenic or non-naturally occurring plants or parts thereof of the present invention exhibit modulated levels of at least one reducing sugar, such as glucose or fructose or a combination thereof, in plant material, such as dried leaves. In certain embodiments, when glucose or fructose levels or a combination thereof are reduced, sucrose levels may be increased.

Preferably, the modulated levels of at least one reducing sugar are observed in at least dried leaves, preferably in fully dried leaves. Preferably, the dried leaves are obtained from leaves obtained from the middle part of the plant. Preferably, there is no or little effect on the phenotype, such as the visual fitness of the plant compared to a control plant. Preferably, there is no or little variation in the total free amino acid content compared to a control plant. In certain embodiments, the acrylamide levels in the smoke obtained by heating the dried leaves or a product obtained therefrom can be modulated.

A further aspect relates to a mutant, non-naturally occurring or transgenic plant or cell as described herein, which is characterized by at least 5% reduced levels of at least one reducing sugar compared to a control plant in which the expression or function of said NtINV or NtINV and NtSUS polypeptide(s) has not been modulated.

In certain embodiments, glucose or fructose levels, or a combination thereof, are reduced by about 30% or more, such as about 40%, or about 50%, or about 60%, or about 70%, or about 80%, or about 90% or more, compared to a control plant.

In certain embodiments, glucose or fructose levels, or a combination thereof, are reduced by about 40% or more, such as about 50%, or about 60%, or about 70%, or about 80%, or about 90% or more, compared to a control plant.

In certain embodiments, the glucose level is reduced by at least 60% or more, or at least 63% or more, and the fructose level is reduced by at least about 40% or more, or at least 63% or more, compared to a control plant.

In certain embodiments, the glucose level is reduced by at least 60% or more, or at least 63% or more, and the fructose level is reduced by at least about 40% or more, or at least 63% or more, and the sucrose level is increased by at least 2-fold, at least 3-fold, or at least 4-fold compared to a control plant.

In certain embodiments, glucose and fructose levels are increased compared to a control plant.

In certain embodiments, the glucose level is increased, the fructose level is increased, and the sucrose level is decreased compared to a control plant.

The increase may be approximately 25%, 50%, 100%, 250%, or 500% or more compared to the control plant. The decrease may be approximately 25%, 50%, or 75% or more compared to the control plant.

Yet another further aspect relates to a dried plant material, such as dried leaf or dried tobacco, obtained or obtainable from a mutant, non-naturally occurring or transgenic plant or cell, wherein the expression of one or more of the NtINV or NtINV and NtSUS polynucleotides described herein, or the function of the NtINV or NtINV and NtSUS polypeptide encoded therefor, is modulated, and wherein the level of glucose, fructose and optionally sucrose is modulated, as discussed above, compared to a control plant.

Embodiments are also directed to compositions and methods for producing mutant, non-naturally occurring, or transgenic plants or plant cells that have been modified to modulate the expression or function of one or more of the NtINV or NtINV and NtSUS polynucleotides or NtINV or NtINV and NtSUS polypeptides described herein, which can result in plants, or plant components (e.g., leaves, such as desiccated leaves), or plant cells with modulated glucose, fructose, and optionally sucrose content.

In one embodiment, the phenotype of the mutant, non-naturally occurring, or transgenic plant is substantially the same as that of the control plant. In one embodiment, the leaf weight of the mutant, non-naturally occurring, or transgenic plant is substantially the same as that of the control plant. In one embodiment, the number of leaves of the mutant, non-naturally occurring, or transgenic plant is substantially the same as that of the control plant. In one embodiment, the leaf weight and the number of leaves of the mutant, non-naturally occurring, or transgenic plant are substantially the same as that of the control plant. In one embodiment, the stem height of the mutant, non-naturally occurring, or transgenic plants is substantially the same as that of the control plants, such as one, two, or three or more months after transplantation to the field or 10, 20, 30, or 36 or more days after topping. For example, the stem height of the mutant, non-naturally occurring, or transgenic plants is not less than the stem height of the control plants. In another embodiment, the chlorophyll content of the mutant, non-naturally occurring, or transgenic plants is substantially the same as that of the control plants. In another embodiment, the stem height of the mutant, non-naturally occurring, or transgenic plants is substantially the same as that of the control plants, and the chlorophyll content of the mutant, non-naturally occurring, or transgenic plants is substantially the same as that of the control plants. In other embodiments, the size or shape or number or color of the leaves of the mutant, non-naturally occurring, or transgenic plants is substantially the same as that of the control plants.

In another aspect, there is provided a method for modulating the amount of at least one reducing sugar in at least a portion of a plant (e.g., leaves, such as dried leaves, or in tobacco), comprising the steps of (i) modulating the expression or function of one or more of the NtINV or NtINV and NtSUS polypeptides described herein (or any combination thereof described herein), wherein preferably the NtINV or NtINV and NtSUS polypeptide(s) is/are encoded by the corresponding NtINV or NtINV and NtSUS polynucleotide(s) described herein; (ii) measuring the level of at least one reducing sugar (e.g. glucose and fructose) and optionally at least one non-reducing sugar, such as sucrose, in at least a portion (e.g. leaves, such as dried leaves, or in tobacco, or in smoke) of the mutant, non-naturally occurring or transgenic plant obtained in step (i); and (iii) identifying a mutant, non-naturally occurring or transgenic plant in which the level of at least one reducing sugar and optionally at least one non-reducing sugar has been modulated compared to a control plant.

In another aspect, there is provided a method of modulating the amount of at least one reducing sugar in at least a portion of a dried plant material, such as a dried leaf, comprising the steps of (i) modulating the expression or function of one or more of the NtINV or NtINV and NtSUS polypeptides (or any combination thereof described herein), wherein preferably the NtINV or NtINV and NtSUS polypeptide(s) is/are encoded by the corresponding NtINV or NtINV and NtSUS polynucleotide(s) described herein; (ii) collecting the plant material, such as one or more leaves, and drying for a period of time; (iii) measuring the level of at least one reducing sugar (e.g., glucose or fructose) and optionally at least one non-reducing sugar, such as sucrose, in at least a portion of the dried plant material obtained in or during step (ii); and (iv) identifying a dried plant material in which the level of at least one reducing sugar and optionally at least one non-reducing sugar has been modulated compared to a control plant.

The increase in expression compared to a control can be from about 5% to about 100%, or the increase is at least 10%, at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 90%, at least 95%, at least 98%, or 100% or more, such as 200%, 300%, 500%, 1000% or more, which includes an increase in transcriptional function, or the expression level of the NtINV polynucleotide or NtINV and NtSUS , or the expression level of the NtINV or NtINV and NtSUS polypeptide, or a combination thereof.

The increase in function or activity compared to a control can be from about 5% to about 100%, or the increase is at least 10%, at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 90%, at least 95%, at least 98%, or 100% or more, such as 200%, 300%, 500%, 1000% or more, which includes an increase in transcriptional function, or the expression level of an NtINV polynucleotide or of NtINV and NtSUS , or the expression level of an NtINV or NtINV and NtSUS polypeptide, or a combination thereof.

The reduction in expression level compared to a control can be from about 5% to about 100%, or the reduction is at least 10%, at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 90%, at least 95%, at least 98% or 100%, which includes a reduction in transcriptional function, or the expression level of the NtINV polynucleotide or NtINV and NtSUS , or the expression level of the NtINV or NtINV and NtSUS polypeptide, or a combination thereof.

The decrease in function or activity compared to a control can be from about 5% to about 100%, or the decrease is at least 10%, at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 90%, at least 95%, at least 98%, or 100%, which includes a decrease in transcriptional function or expression level of an NtINV or NtINV and NtSUS polynucleotide, or expression level of an NtINV or NtINV and NtSUS polypeptide, or a combination thereof.

The polynucleotides and recombinant constructs described herein can be used to modulate the expression or function or activity of the NtINV or NtINV and NtSUS polynucleotides or the NtINV or NtINV and NtSUS polypeptides described herein in a plant species of interest, preferably tobacco.

A variety of polynucleotide-based methods can be used to increase gene expression levels in plants and plant cells. As an example, a construct, vector, or expression vector compatible with a plant to be transformed can be produced that comprises a gene of interest together with an upstream promoter capable of driving overexpression of the gene in a plant or plant cell. Exemplary promoters are described herein. Following transformation and growth under appropriate conditions, the promoter can drive expression to modulate levels of NtINV or NtINV and NtSUS in a plant or a particular tissue thereof. In one exemplary embodiment, a vector is generated that carries one or more of the NtINV or NtINV and NtSUS polynucleotides described herein (or any combination thereof as described herein) to drive overexpression of a gene in a plant or plant cell. The vector carries a suitable promoter, such as the cauliflower mosaic virus CaMV 35S promoter, upstream of the transgene, which drives its constitutive expression in all plant tissues. The vector also carries an antibiotic resistance gene to allow selection of transformed calli and cell lines.

Expression of sequences from promoters can be enhanced by incorporating expression control sequences, which are well known in the art. Separate mention should be made of signals associated with aging and signals that are active during the drying procedure.

Various embodiments are thus directed to methods of modulating the expression level of one or more NtINV or NtINV and NtSUS polynucleotides described herein (or any combination thereof as described herein) by integrating multiple copies of the NtINV or NtINV and NtSUS polynucleotide into a plant genome, wherein the methods comprise transforming a plant host cell with an expression vector that comprises a promoter operably linked to one or more NtINV or NtINV and NtSUS polynucleotides described herein. The polypeptide encoded by the recombinant polynucleotide may be a native polypeptide or may be heterologous with respect to the cell.

In one embodiment, the plant for use in the present invention is a plant that has been flue-dried, since such plants are characterized by a high content of reducing sugars (when grown in the field, more than about 14% of the dry weight at the end of drying). Mutant, transgenic, or non-naturally occurring plants or parts thereof that have been flue-dried may be characterized by a content of reducing sugars that is less than about 14% of the dry weight when grown in the field at the end of drying, such as less than about 10% of the dry weight when grown in the field at the end of drying, or less than about 5% of the dry weight when grown in the field at the end of drying, or less than about 1% of the dry weight when grown in the field at the end of drying.

In one embodiment, the plant for use in the present invention is a plant that has been sun-dried (since such plants have a reducing sugar content greater than about 6.8% of the dry weight when grown in the field at the end of drying). Mutant, transgenic, or non-naturally occurring plants or parts thereof that have been sun-dried may have a reducing sugar content of less than about 5% of the dry weight when grown in the field at the end of drying, such as less than about 2.5% of the dry weight when grown in the field at the end of drying, or less than about 1% of the dry weight when grown in the field at the end of drying.

In one embodiment, the plant for use in the present invention is an air-dried plant. Such plants are characterized by a reducing sugar content of greater than about 1.7% of the dry weight when grown in the field at the end of drying. Mutant, transgenic, or non-naturally occurring plants or parts thereof that are sun-dried may be characterized by a reducing sugar content of less than about 1.5% of the dry weight when grown in the field at the end of drying, such as less than about 1% of the dry weight when grown in the field at the end of drying, or less than about 0.5% of the dry weight when grown in the field at the end of drying.

In certain embodiments, it is preferred to use flue-dried or sun-dried plants.

9. Selection

A plant carrying a mutant allele of one or more NtINV or NtINV and NtSUS polynucleotides described herein (or any combination thereof described herein) can be used in a plant breeding program to create useful lines, varieties, and hybrids. In particular, the mutant allele can be introgressed into the commercially valuable varieties described above. Thus, plant breeding methods are provided that involve crossing a mutant plant, a non-naturally occurring plant, or a transgenic plant as described herein with a plant having different genetic characteristics. The method can further comprise crossing the progeny plant with another plant, and optionally repeating the cross until a progeny with the desired genetic traits or genetic background is obtained. One purpose for which such breeding methods are used is to introduce a desired genetic trait into other varieties, breeding lines, hybrids or cultivars, especially those of commercial interest. Another purpose is to facilitate the accumulation of genetic modifications of various genes in individual varieties, lines, hybrids or cultivars of plants. Intraspecific as well as interspecific crosses are envisaged. The offspring plants that result from such crosses, also called breeding lines, are examples of non-naturally occurring plants according to the present invention.

In one embodiment, a method for producing a non-naturally occurring plant is provided, comprising: (a) crossing a mutant or transgenic plant with a second plant to produce a seed of a progeny tobacco plant; (b) growing the seed of the progeny tobacco plant under conditions suitable for plant growth to produce a non-naturally occurring plant. The method may further comprise: (c) crossing a previous generation of the non-naturally occurring plant with itself or with another plant to produce a seed of a progeny tobacco plant; (d) growing the seed of the progeny tobacco plant from step (c) under conditions suitable for plant growth to produce additional non-naturally occurring plants; and (e) repeating the crossing and growing steps of (c) and (d) several times to produce further generations of non-naturally occurring plants. The method may optionally comprise, prior to step (a), the step of obtaining a parent plant that comprises the characterized genetic characteristics and that is not identical to the mutant or transgenic plant. In some embodiments, depending on the breeding program, the crossing and growing steps are repeated 0 to 2 times, 0 to 3 times, 0 to 4 times, 0 to 5 times, 0 to 6 times, 0 to 7 times, 0 to 8 times, 0 to 9 times, or 0 to 10 times to obtain generations of non-naturally occurring plants. Backcrossing is an example of such a method, in which an offspring is crossed with one of its parents or with another plant genetically similar to its parent, to obtain a progeny plant in the next generation that has genetic characteristics closer to one of the parents. Plant breeding techniques, in particular plant breeding, are well known and can be used in the methods of the present invention. The present invention further provides non-naturally occurring plants obtained using these methods. Certain embodiments eliminate the step of selecting the plant.

In some embodiments of the methods described herein, lines resulting from selection and screening for variant genes are evaluated in the field using standard field procedures. Control genotypes, including the genotype of the original non-mutagenesis parent, are provided and plant locations are arranged in the field according to a randomized complete block design or other appropriate field design. For tobacco, standard agronomic techniques are used, such as harvesting, weighing, and sampling for chemical and other conventional testing before and during curing. Statistical analysis of the data is performed to confirm the similarity of the selected lines to the parental line. Cytogenetic analyses of the selected plants are optionally performed to confirm chromosome complement relationships and chromosome pairing.

DNA fingerprinting, single nucleotide polymorphism, microsatellite markers, or similar technologies may be used in a marker-assisted selection (MAS) breeding program to transfer or breed mutant alleles of a gene into other tobacco plants, as described herein. For example, a breeder may create segregating populations by hybridizing a genotype containing a mutant allele with an agronomically desirable genotype. Plants from the F2 or backcross generations may be screened using a marker derived from the genomic sequence or a fragment thereof using one of the techniques described herein. Plants identified as having the mutant allele may be backcrossed or selfed to create a second population for screening. Depending on the expected mode of inheritance or the MAS technology used, selected plants may need to be self-pollinated before each backcross cycle to facilitate detection of the desired individual plants. The backcross or other selection procedure can be repeated until the desired phenotype of the recurrent parent is restored.

According to the present invention, in a breeding program, successful crosses produce F1 plants that are fertile. The selected F1 plants can be crossed with one of the parents, and the first generation of plants obtained from the backcross can be self-pollinated to produce a population that is again screened for expression of the variant gene (e.g., a null version of the gene). The process of backcrossing, self-pollination, and screening is repeated, for example, at least 4 times until a final screening yields a plant that is fertile and sufficiently similar to the recurrent parent. This plant is optionally self-pollinated and the progeny are then screened again to confirm that the plant exhibits expression of the variant gene. In some embodiments, the F2 generation population of plants is screened for expression of the variant gene, e.g., a plant that does not express the polypeptide due to the absence of the gene is identified according to standard methods, e.g., using PCR techniques using primers designed based on the polynucleotide sequence information for the polynucleotide(s) described herein (or any combination thereof described herein).

Hybrid tobacco varieties can be produced by preventing self-pollination of female parent plants (i.e., parent forms) of the first variety, allowing pollen from male parent plants of the second variety to fertilize the female parent plants and allowing F1 hybrid seeds to form on the female plants. Self-pollination of female plants can be prevented by emasculating flowers at an early stage of flower development. Alternatively, pollen formation on female parent plants can be prevented by using some form of male sterility. For example, male sterility can be achieved through cytoplasmic male sterility (CMS) or transgenic male sterility, wherein the transgene suppresses microsporogenesis or pollen formation or causes self-incompatibility. Female parents containing CMS are particularly useful. In embodiments in which the female parents are characterized by CMS, pollen is collected from male fertile plants and manually applied to the stigmas of female parents with CMS and the resulting F1 seeds are collected.

The varieties and lines described herein can be used to form F1 single hybrids of tobacco. In such embodiments, the parent variety plants can be grown as substantially uniform adjacent populations to facilitate natural cross-pollination of the female parent plants with the male parent plants. The F1 seeds produced on the female parent plants are selectively collected by conventional means. It is also possible to grow two varieties of the parent plant together and collect a mixture of the F1 hybrid seeds produced on the female and seeds produced without the male as a result of self-pollination. Alternatively, a trilinear cross can be performed where an F1 single hybrid is used as a female and crossed with another male. As another alternative, double cross hybrids can be created where the F1 progeny of two different single hybrids are crossed with themselves.

A population of mutant, non-naturally occurring, or transgenic plants can be screened or selected for those members of the population that have a desired trait or phenotype. For example, a population of progeny of a single transformation line can be screened for those plants that have the desired level of expression or function of the polypeptide(s) encoded by it. Physical and biochemical methods can be used to detect expression levels or activity. These include Southern analysis or PCR amplification to detect the polynucleotide; Northern blotting, RNase S1 protection assay, primer extension assay, or RT-PCR amplification to detect RNA transcripts; enzymatic assays to detect enzymatic or ribozyme function of polypeptides and polynucleotides; and polypeptide gel electrophoresis, Western blotting, immunoprecipitation, and enzyme immunoassays to detect polypeptides. Other techniques such as in situ hybridization, enzymatic staining and immunostaining, and enzymatic assays can also be used to detect the presence or expression or function or activity of NtINV or NtINV and NtSUS polypeptides or polynucleotides.

Described herein are mutant, non-naturally occurring or transgenic plant cells and plants comprising one or more recombinant polynucleotides, one or more polynucleotide constructs, one or more double-stranded RNAs, one or more conjugates or one or more vectors/expression vectors.

10. Modification of other genes

Without limitation, the plants and parts thereof described herein can be modified before or after modulating the expression, function, or activity of one or more NtINV or NtINV and NtSUS polynucleotides or NtINV or NtINV and NtSUS polypeptides of the present invention.

One or more of the following additional genetic modifications may be present in mutant, non-naturally occurring or transgenic plants or parts thereof.

One or more genes that are involved in the conversion of nitrogen metabolism intermediates may be modified, resulting in lower levels of at least one tobacco-specific nitrosamine (TSNA). Non-limiting examples of such genes include genes encoding nicotine demethylase, such as CYP82E4, CYP82E5 and CYP82E10, described in WO2006/091194, WO2008/070274, WO2009/064771 and WO2011/088180, as well as nitrate reductase, described in WO2016/046288.

One or more genes that are involved in the uptake of heavy metals or the transport of heavy metals may be modified, resulting in a lower heavy metal content. Non-limiting examples include the multidrug resistance-associated polypeptide family, the cation diffusion mediator (CDF) family, the Zrt-, Irt-like polypeptide (ZIP) family, the cation exchanger (CAX) family, the copper transporter (COPT) family, the heavy metal ATPase family (e.g., HMA, as described in WO2009/074325 and WO2017/129739), the natural resistance-associated macrophage polypeptide homologue (NRAMP) family, and other members of the ATP-binding cassette (ABC) transporter family (e.g., MRP), as described in WO2012/028309, which are involved in the transport of heavy metals such as cadmium.

Other illustrative modifications may result in plants with modulated expression or function of isopropyl malate synthase, resulting in an altered sucrose ester composition that can be used to alter the aroma profile (see WO2013/029799).

Other illustrative modifications may result in plants with modulated expression or function of threonine synthase, whereby methional levels may be modulated (see WO2013/029800).

Other illustrative modifications may result in plants with modulated expression or function of one or more of neoxanthine synthase, lycopene beta-cyclase and 9-cis-epoxycarotenoid dioxygenase to modulate beta-damascenone content to alter the flavour profile (see WO2013/064499).

Other illustrative modifications may result in plants with modulated expression or function of members of the CLC family of chloride channels to modulate nitrate levels therein (see WO2014/096283 and WO2015/197727).

Other illustrative modifications may result in plants having modulated expression or function of one or more asparagine synthetases to modulate asparagine levels in the leaf and provide modulated acrylamide levels in the aerosol generated by heating or burning the leaf (see WO2017/129739).

Other illustrative modifications may result in plants with modulated protease activity during drying (see WO2016/009006).

Other illustrative modifications can produce plants with reduced nitrate levels by altering the expression of the nitrate reductase gene (e.g.,Nia2) or activity protein encoded by it (see WO2016/046288).

Other illustrative modifications can result in plants with modified alkaloid levels by altering the expression levels of putative ABC-2 transporter genes.NtABCGl-T And NtABCGl-Sor activity proteins encoded by them (see WO2019/086609).

Other illustrative modifications can result in plants with modulated time to flowering by altering the gene expression of genes encoding Terminal Flower 1 (TFL1) or the activity protein encoded therefor (see WO2018/114641). Other illustrative modifications may result in plants with modulated expression or function of one or more asparagine synthetases to modulate asparagine levels in the leaf and provide modulated acrylamide levels in the aerosol generated by heating or burning the leaf (see WO2017/042162).

Examples of other modifications include modulation of herbicide tolerance, for example glyphosate is the active ingredient in many broad-spectrum herbicides. Glyphosate-resistant transgenic plants have been developed by transferring the aroA gene (glyphosate EPSP synthetase from Salmonella typhimurium and E. coli ). Sulfonylurea-resistant plants have been produced by transforming a mutant ALS (acetolactate synthetase) gene from Arabidopsis. Photosystem II polypeptide OB from a mutant Amaranthus hybridus has been transferred into plants to produce atrazine-resistant transgenic plants; and bromoxynil-resistant transgenic plants have been produced by inserting the bxn gene from the bacterium Klebsiella pneumoniae.

Other examples of modifications result in plants that are resistant to insects. ToxinsBacillus thuringiensis(Bt) may provide an effective way delaying the emergence of Bt-resistant pests, as recently shown on broccoli, where the Bt genes arecry1Acand cry1C as part of the "pyramid" provided control of the cabbage moth, resistant to any individual polypeptide, and significantly delayed evolutionary development of resistant insects.

Another illustrative modification results in plants that are resistant to diseases caused by pathogens (e.g., viruses, bacteria, fungi). Plants expressing the Xa21 gene (resistance to bacterial blight) have been developed with plants expressing both the Bt fusion gene and the chitinase gene (resistance to yellow moth and tolerance to rhizoctonia).

Another illustrative modification results in an adjustment of reproductive capacity, such as male sterility.

Another illustrative modification results in plants that are tolerant to abiotic stress (e.g. drought, temperature change, salinity), and tolerant transgenic plants have been produced by transferring the enzyme acylglycerol phosphate acyltransferase from Arabidopsis; genes encoding mannitol dehydrogenase and sorbitol dehydrogenase, which are involved in the synthesis of mannitol and sorbitol, improve drought tolerance.

Another illustrative modification results in plants in which the activity of one or more nicotine N-demethylases is modulated such that the levels of nornicotine and nornicotine metabolites that are formed during drying can be modulated (see WO2015169927).

Other exemplary modifications may result in plants with improved characteristics of storage polypeptides and oils, plants with increased efficiency of photosynthesis, plants with a long shelf life, plants with increased carbohydrate content, and plants that are resistant to fungi. Transgenic plants in which the expression levels of S-adenosyl-L-methionine (SAM) and/or cystathionine gamma synthase (CGS), or a combination thereof, are modulated are also provided.

One or more genes that are involved in the nicotine synthesis pathway can be modified to produce plants or plant parts that produce modulated levels of nicotine when dried. The nicotine synthesis genes can be selected from the group consisting of: A622, BBLa, BBLb, JRE5L1, JRE5L2, MATE1, MATE 2, MPO1, MPO2, MYC2a , MYC2b, NBB1, nic1, nic2, NUP1, NUP2, PMT1, PMT2, PMT3, PMT4 and QPT or a combination of one or more of them.

One or more genes that are involved in regulating the amount of one or more alkaloids can be modified to produce plants or plant parts that produce modulated levels of the alkaloid when dried. The genes that provide control of the alkaloid level can be selected from the group consisting of BBLa , BBLb, JRE5L1, JRE5L2, MATE1, MATE 2 , MYC2a, MYC2b , nic1, nic2, NUP1 and NUP2 or a combination of two or more of them.

One or more such traits can be introgressed into mutant, non-naturally occurring or transgenic plants of another variety, or can be directly transformed into them.

In various embodiments, mutant plants, non-naturally occurring plants, or transgenic plants, as well as biomass, are provided in which the expression level of one or more polynucleotides according to the present invention are modulated to thereby modulate the level of the polypeptide(s) encoded thereby.

11. Consumer products

Parts of the plants described herein, in particular the blade and midrib of these plants, can be incorporated into or used in the manufacture of various consumer products, including, but not limited to, aerosol-generating materials, aerosol-generating devices, smoking articles, smoking products, smokeless products, medicinal or cosmetic products, intravenous preparations, tablets, powders, and tobacco products. Examples of aerosol-generating materials include tobacco compositions, tobacco types, tobacco extract, shredded tobacco, shredded filler, cured tobacco, blown tobacco, homogenized tobacco, reconstituted tobacco, and pipe tobacco types. Smoking articles and smoking articles are typically aerosol-generating devices. Examples of smoking articles or smoking articles include cigarettes, cigarillos, and cigars. Examples of smokeless products include chewing tobacco and snuff. In certain aerosol-forming devices, instead of combustion, the tobacco composition or other aerosol-forming material is heated by one or more electrical heating elements to produce an aerosol. In another type of heated aerosol-forming device, the aerosol is produced by transferring heat from a combustible fuel element or heat source to a physically separate aerosol-forming material, which may be located within, around, or downstream of the heat source. Smokeless tobacco products and various tobacco-containing aerosol-forming materials may contain tobacco in any form, including dried particles, pieces, granules, powders, or suspensions coated on, mixed with, surrounded by, or otherwise combined with other ingredients in any format, such as flakes, films, tablets, foams, or pellets. The term "smoke" is used to describe the type of aerosol that is produced by smoking articles such as cigarettes or by burning an aerosol-forming material.

In one embodiment, dried plant material from mutant, transgenic and non-naturally occurring plants described herein is also provided. Methods for drying green tobacco leaves are known to those skilled in the art and include, but are not limited to, air drying, fire drying, flue drying and solar drying as described herein.

In another embodiment, tobacco products are described, including tobacco-containing aerosol-forming materials, comprising plant material such as leaves, preferably dried leaves, from mutant tobacco plants, transgenic tobacco plants or non-naturally occurring tobacco plants described herein. The tobacco products described herein may be blended tobacco products that may further comprise unmodified tobacco.

12. Products and methods for crop management and agriculture

For mutant, non-naturally occurring or transgenic plants, there may be other applications in, for example, agriculture.

The present invention also provides methods for producing seeds, comprising cultivating a mutant plant, a non-naturally occurring plant or a transgenic plant described herein and collecting seeds of the cultivated plants. The seeds of the plants described herein can be conditioned and packaged in a packaging material by means known in the art to produce an industrial article. The packaging material, such as paper or fabric, is well known in the art. The seed packaging may have a label, such as a mark or label attached to the packaging material, a label printed on the package, which describes the origin of the seeds contained therein.

Compositions, methods and kits for genotyping plants for identification, selection or breeding may include means for detecting the presence of NtINV or NtINV and NtSUS polynucleotide(s) in a polynucleotide sample. Accordingly, a composition is described comprising one or more primers for specifically amplifying at least a portion of one or more of the NtINV or NtINV and NtSUS polynucleotides, and optionally one or more probes, and optionally one or more reagents for carrying out the amplification or detection.

Accordingly, gene-specific oligonucleotide primers or probes are disclosed that comprise about 10 or more contiguous polynucleotides corresponding to the NtINV or NtINV and NtSUS polynucleotide(s) described herein. The primers or probes may comprise or consist of about 15, 20, 25, 30, 40, 45, or 50 or more contiguous polynucleotides that hybridize (e.g., specifically hybridize) to the NtINV or NtINV and NtSUS polynucleotide(s) described herein. In some embodiments, the primers or probes may comprise or consist of about 10-50 contiguous nucleotides, about 10-40 contiguous nucleotides, about 10-30 contiguous nucleotides, or about 15-30 contiguous nucleotides that can be used in sequence-dependent methods for identifying (e.g., Southern blot hybridization) or isolating a gene (e.g., in situ hybridization of bacterial colonies or bacteriophage plaques) or detecting a gene (e.g., as one or more amplification primers in amplification or detection). One or more specific primers or probes can be designed and used to amplify and detect a portion or all of the polynucleotide(s). As a specific example, two primers can be used in a PCR protocol to amplify a polynucleotide fragment. PCR can also be performed using one primer that is derived from a polynucleotide sequence and a second primer that hybridizes to a sequence upstream or downstream of the polynucleotide sequence, such as a promoter sequence, the 3' end of a precursor mRNA, or a sequence derived from a vector. Examples of thermal and isothermal techniques useful for in vitro amplification of polynucleotides are well known in the art. The sample can be derived from or obtained from a plant, plant cell, or plant material, or a tobacco product made from a plant, plant cell, or plant material, as described herein.

In a further aspect, there is also provided a method for detecting the NtINV or NtINV and NtSUS polynucleotide(s) described herein (or any combination thereof described herein) in a sample, comprising the steps of (a) obtaining a sample comprising or suspected of comprising the polynucleotide; (b) contacting said sample with one or more primers or one or more probes to specifically detect at least a portion of the NtINV or NtINV and NtSUS polynucleotide(s); and (c) detecting the presence of an amplification product, wherein the presence of an amplification product indicates the presence of the NtINV or NtINV and NtSUS polynucleotide(s) in the sample. In a further aspect, there is also provided the use of one or more primers or probes to specifically detect at least a portion of the NtINV or NtINV and NtSUS polynucleotide ( s ). Also provided are kits for detecting at least a portion of the NtINV or NtINV and NtSUS polynucleotide(s), which comprise one or more primers or probes for specifically detecting at least a portion of the NtINV or NtINV and NtSUS polynucleotide(s). The kit may comprise reagents for amplifying the polynucleotide, such as PCR reagents, or reagents for a hybridization probe detection technology such as Southern blotting, Northern blotting, in situ hybridization, or microarray. The kit may comprise reagents for antibody binding detection technology such as Western blotting, ELISA, SELDI mass spectrometry, or test strips. The kit may comprise reagents for DNA sequencing. The kit may comprise reagents and instructions for using the kit.

In some embodiments, the kit may contain instructions for one or more of the described methods. The described kits may be useful for determining genetic identity, phylogenetic studies, genotyping, haplotyping, pedigree analysis, or plant breeding, in particular with the quantitative assessment of codominant traits.

The present invention also provides a method for genotyping a plant, plant cell, or plant material comprising the NtINV or NtINV and NtSUS polynucleotide described herein. Genotyping provides a means of distinguishing homologs of a pair of chromosomes and can be used to distinguish segregants in a plant population. Molecular marker-based methods can be used for phylogenetic studies characterizing genetic relationships between crop varieties, identifying hybrids or somatic hybrids, localizing chromosomal segments that affect monogenic traits, cloning based on genetic maps, and studying quantitative inheritance. A particular genotyping method can use any number of molecular marker assays, including those based on amplified fragment length polymorphism (AFLP). AFLP is the result of allelic differences between amplified fragments caused by polynucleotide variability. Thus, the present invention further provides methods for monitoring the segregation of one or more genes or polynucleotides, as well as chromosomal sequences genetically linked to these genes or polynucleotides, using techniques such as AFLP analysis.

13. Tobacco extracts

This document also discloses methods for producing liquid tobacco extract and liquid tobacco extract obtained by such method(s).

The specific extraction temperature is selected for the tobacco starting material, preferably based on at least the reducing sugar content and, optionally, the nicotine content of the tobacco starting material(s). The extraction temperature(s) are typically selected within the range of about 100 degrees Celsius to about 160 degrees Celsius. The duration of the heating step can optionally be adjusted to provide a degree of control over the composition of the extract obtained from the tobacco starting material(s). Preferably, the tobacco starting material(s) is heated at the extraction temperature for at least about 90 minutes, more preferably at least about 120 minutes. The heating step is typically carried out in an inert atmosphere. Preferably, a stream of inert gas, such as nitrogen, is passed through the tobacco starting material during the heating step. The volatile tobacco compounds are released into the inert gas stream during the heating step, so that the inert gas acts as a carrier for the volatile components. The inert gas stream may have a flow rate of at least about 25 liters per minute, more preferably at least about 30 liters per minute. A relatively high flow rate of the inert gas may advantageously increase the extraction efficiency from the tobacco starting material. Optionally, the heating step may be carried out in a vacuum. Suitable heating methods for carrying out the heating of the tobacco starting material are known to those skilled in the art and include: dry distillation, hydrodistillation, vacuum distillation, equilibrium distillation and thin film hydrodistillation.

If the volatile compounds are collected by absorption into a liquid solvent, the step of forming a mixed liquid tobacco extract may include drying a solution of the volatile compounds in the liquid solvent to concentrate the solution. Drying may be accomplished using any suitable means, including, but not limited to, dehydration, molecular sieves, freeze-drying, phase separation, distillation, membrane permeation, controlled water crystallization and filtration, reverse hygroscopicity, ultracentrifugation, liquid chromatography, reverse osmosis, or chemical drying.

A liquid tobacco extract is particularly suitable for producing a composition or formulation or gel composition for use in an aerosol generating system. An aerosol generating system is disclosed that contains the composition or formulation or gel composition. In such an aerosol generating system, the composition or formulation or gel is typically heated in an aerosol generating device, such as a device containing a heating element that interacts with the composition or formulation or gel containing the liquid tobacco extract to form an aerosol. During use, volatile compounds are released from the composition by heat transfer and are entrained in air drawn through the aerosol generating device. As the released compounds cool, they condense to form an aerosol that is inhaled by the user.

The present invention is further described in the examples below, which are presented to describe the present invention in more detail. These examples, which set forth the preferred mode presently contemplated for carrying out the present invention, are intended to illustrate and not to limit the present invention.

EXAMPLES

Example 1. Materials and methods

DNA extraction and genotyping of plants

Leaf samples were extracted using BioSprint 96 (Qiagen, Hilden, Germany) together with the BioSprint 96 DNA plant kit (Qiagen, Hilden, Germany). The DNA samples were used in a TaqMan reaction to determine the genotype of the plant. The Taqman reaction was performed using an ABI PRISM 7900HT sequencer (Applied Biosystems, Life Technologies, Foster City, CA, USA) and TaqMan Fast Advanced master mix (Applied Biosystems, Foster City, CA, USA).

Measurement of free amino acid content

The amino acid content can be measured using various methods known in the art. One such method is the method of MP 1471 rev 5 2011, Resana, Italy: Chelab Silliker S.r.l, Mérieux NutriSciences Company. To detect the amino acids in the dried leaves of the plant, after removing the midrib, the dried leaves are dried, if necessary, at 40°C for 2-3 days. The tobacco material is then ground into a fine powder (~100 μM) before being analyzed for amino acid content. Another method for measuring the amino acid content of plant material is described in UNI EN ISO 13903:2005. The measurement of free amino acids can be carried out in accordance with UNI EN ISO 13903:2005.

Measurement of reducing sugars content

Reducing sugars can be measured using a segmented flow colorimetric method developed for the analysis of tobacco samples, adapted by Skalar Instrument Co. (West Chester, PA) and described in Tobacco Science 20: 139-144 (1976). The measurement of reducing sugars is also described in Coresta Recommended Method 38, CRM38, CRM and ISO 15154:2003. To determine reducing sugars in dried leaves, the dried midrib-removed lamellae are dried at 40°C for 2-3 days if necessary. The tobacco material is then ground to a fine powder (~100 µm) before analysis for reducing sugars. Reducing sugars are measured in accordance with ISO 15154:2003.

Example 2. Expression analysis NtINV4-S And NtINV4-T in plant tissues

Table 1 shows that NtINV4-S and NtINV4-T are expressed in whole plant tissues, particularly in the petal, and also in the immature flower, sepal, lower leaf, midleaf and upper leaf to a lesser extent in field-grown Virginia tobacco plants. In contrast, NtINV3-S and NtINV3-T are expressed at very low levels in the tissues examined, with measurable expression levels, although very low, present only in the immature flower of field-grown Virginia tobacco plants.

Example 3. Dynamics of reducing sugars (glucose and fructose) during flue-curing of Virginia tobacco

During flue-cured Virginia tobacco, the content of reducing sugars, mainly glucose and fructose, increases strongly in yellowed leaves, reaching a maximum level after one or two days (see Figure 1A). Interestingly, the expression level of NtINV4-S and NtINV4-T increases approximately 2-fold (in log2), reaching a plateau after 1 or 2 days (see Figure 1A). NtINV4-T is expressed to a higher extent during drying than NtINV4-S (see Figure 1B and Table 2). Both copies of NtINV3 are not significantly expressed in the dried leaf (see Figure 2B) or the green leaf (see Table 1). As a control, SAG12 , a general marker of plant senescence, is fully expressed after one day of drying (see Figure 1C).

Example 4. Dynamics of reducing sugars (glucose and fructose) in dark tobacco during air drying

To determine whether the induction of NtINV4-S and NtINV4-T is not specific to Virginia tobacco, expression data in dark tobacco were examined. In this case, leaves were air-dried. During the first 120 h, samples were collected at 24, 48, 96, and 120 h, frozen, lyophilized, and subjected to metabolomic analyses. In Virginia tobacco leaf, glucose and fructose (reducing sugars) were observed to increase 4-6-fold from 0 h to 120 h of drying (end of yellowing phase) (see Figure 2). In fully dried leaves (approximately 10 days after the start of drying), glucose and fructose levels were reduced in the leaf matrix as in flue-cured tobacco (compare Figures 1 and 2), but to a greater extent. Sucrose levels remained nearly stable during the first 120 hours of drying, suggesting that dark tobacco maintains a reserve of sucrose molecules during leaf yellowing. At the end of drying, sucrose is completely hydrolyzed and thus likely completely metabolized (see Figure 2C).

For the same samples, frozen leaf material is used to isolate RNA for expression analysis of NtINV3-S , NtINV3-T , NtINV4-S and NtINV4-T . As observed for flue-cured tobacco, the NtINV3 genes are not expressed in the harvested leaf and are not upregulated during air-curing of dark tobacco. On the other hand, the activation level of NtINV4-S and NtINV4-T increases by a factor of more than 10 and reaches a maximum after 5 days of drying (120 hours, see Table 2). As observed for flue-cured tobacco, the activation of SAG12 occurs more rapidly and then decreases after 5 days of drying (120 hours).

Example 5. Silencing NtINV4 in flue-cured tobacco

Silencing of NtINV4 was investigated in flue-cured tobacco to determine whether these genes contribute to the reduction of reducing sugars in flue-cured tobacco leaves. A specific DNA fragment (SEQ ID NO: 9) within the coding sequence of both NtINV4-S and NtINV4-T was cloned into the strong mirabilis mosaic virus (MMV) promoter in the GATEWAY vector. The NtINV4 gene fragment is flanked between MMV and the 3' terminator sequence of the nos gene of Agrobacterium tumefaciens nopaline synthase. Tobacco line K326 was transformed using standard Agrobacterium-mediated transformation protocols. The results are shown in Figure 3 for INV4 expression levels for four controls and four 35S:INV4-RNAi transgenic lines. To ensure selection of plants with low levels of reducing sugars, leaves of individual T0 plants and corresponding control lines were analyzed after 48 h of drying to determine the effect on reducing sugars. The best T0 lines, showing the lowest level of NtINV4 expression compared to the control lines, were selected using qPCR. Seeds were collected from these best T0 lines. Manipulation of NtINV4 genes (e.g., either using a constitutive promoter or a specific senescence-inducible promoter such as SAG12 or E4) can alter the chemistry of dried tobacco leaves. Similarly, knockout of NtINV4 genes using a genome editing strategy such as CRISPR or mutant selection can alter the amino acid chemistry of leaves of major commercial tobacco varieties.

Example 6. Analysis of glucose, fructose and sucrose levels in fully dried leaves of four controls and four 35S:INV4-RNAi transgenic lines

Five midplant leaves from the four controls and the four 35S:INV4-RNAi transgenic lines described in Figure 3 were harvested at maturity and subjected to flue-drying. Sugars (glucose, fructose, and sucrose) were analyzed in the fully dried leaves. The data presented in Figure 4 show a strong and significant reduction in glucose and fructose content in INV4-silenced plants. Glucose and fructose levels were reduced by approximately 63% and approximately 43%, respectively. Interestingly, sucrose levels were significantly increased by 4.1-fold in the 35S:INV4-RNAi lines, thus suggesting that part of the sucrose pool that accumulates during leaf drying is not hydrolyzed by the INV4 genes in the silenced lines. No effect on plant visual fitness and no difference in total free amino acid content was observed between control and 35S:INV4-RNAi lines.

Knockout or silencing of NtINV4-S and NtINV4-T can reduce the reducing sugars in desiccated leaves. For a further reduction of reducing sugars, a combination of knockout or silencing of NtSUS and NtINV can be considered. To increase the pool of reducing sugars in desiccated leaves, overexpression of NtINV4-S or NtINV4-T or their combination can be considered using a senescence-inducible promoter such as SAG12 or E4 (use of a constitutive promoter can alter plant metabolism during the vegetative stage).

Example 7. Identification of SUS genes after curing in Burley, Virginia and Oriental tobacco leaf

To identify key functions contributing to sucrose metabolism during early stages of leaf curing in Burley, Virginia, and Oriental tobacco, a prevalence of function analysis was performed for genes upregulated in cured leaves after 48 h of curing compared to mature leaves at harvest (log2-fold change >2, adjusted p-value <0.05) in Burley, Virginia, and Oriental tobacco. Genes involved in reducing sugar production that were active after 48 h of curing independent of curing types and tobacco cultivars were identified. Tobacco genes involved in reducing sugar production were identified.

The key genes directly involved in the production of reducing sugars during the early stages of drying in leaves belong to the SUS gene family. SUS is likely to be the key enzyme for triggering the accumulation of reducing sugars in dried detached leaves.

The tobacco genome was found to have 12 NtSUS gene products distributed across 6 families, with one copy of S and one copy of T from each ancestor: NtSUS1-S (SEQ ID NO: 10) , NtSUS1-T (SEQ ID NO: 12) , NtSUS2-S (SEQ ID NO: 14), NtSUS2-T (SEQ ID NO: 16) , NtSUS3-S (SEQ ID NO: 18) , NtSUS3-T (SEQ ID NO: 20) , NtSUS4-S (SEQ ID NO: 22) , NtSUS4-T (SEQ ID NO: 24) , NtSUS5-S (SEQ ID NO: 26) , NtSUS5-T (SEQ ID NO: 28) , NtSUS6-S (SEQ ID NO: 30) and NtSUS6-T (SEQ ID NO: 32).

SUS transcripts were derived from the genomic sequences NtSUS2-S (SEQ ID NO: 14), NtSUS3-S (SEQ ID NO: 18), NtSUS3-T (SEQ ID NO: 20) and NtSUS4-S (SEQ ID NO: 22). The expression of these genes was increased during leaf drying (senescence) as shown in Table 3. This confirms that S copies in particular are involved in the chemical modification of leaves subjected to early stages of drying and, in this particular case, in the increase in glucose and fructose levels.

Although low reducing sugar levels were found in cured Burley tobacco leaves compared to Virginia and Oriental tobacco, NtSUS genes are nevertheless upregulated in Burley tobacco (see Table 3), likely as a constitutive response to provide an available carbon source for amino acid synthesis during the early curing phase.

In both Burley (BU) and Virginia (FC) tobacco , NtSUS1-S and NtSUS1-T , which are not expressed during the early stages of drying (see Table 3), are particularly expressed in the root and stem, suggesting a possible specific function in these tissues to supply carbohydrates for cell wall synthesis or to provide carbon resources under anoxic conditions (see Table 4). On the other hand , NtSUS3-S, NtSUS3-T, NtSUS4-S , which are induced during the early stages of leaf drying, are also expressed in all organs, whereas NtSUS2-S and NtSUS2-T are expressed predominantly in immature flowers and petals. NtSUS5-S, NtSUS5-T, NtSUS6-S and NtSUS6-T are expressed at low levels in all plant tissues analyzed (see Table 4).

To increase the pool of reducing sugars in dried leaves, overexpression could be considered.NtSUS2-S,NtSUS3-S, NtSUS3-T or NtSUS4-Sor a combination thereof using a senescence-inducible promoter such as SAG12 or E4 (using a constitutive promoter can significantly alter plant metabolism). On the other hand, performing a knockoutNtSUS2-S,NtSUS3-S, NtSUS3-Tand/orNtSUS4-Smay contribute to the reduction of reducing sugars in dried leaves.

Example 8. Expression Silencing NtSUS in a Virginia tobacco leaf

Silencing of NtSUS was investigated to determine whether these genes contribute to the reduction of reducing sugars in cured Virginia tobacco leaves. A specific DNA fragment within the coding sequence of both NtSUS was cloned with the strong promoter of the night beauty mosaic virus (MMV) into the GATEWAY vector. The NtSUS gene fragment is flanked between MMV and the 3' terminator sequence of the nos gene of the nopaline synthase gene of Agrobacterium tumefaciens .

To ensure selection of plants with low levels of reducing sugars, leaves of individual T0 plants and corresponding control lines were analyzed after 60 h of drying to determine the effect on reducing sugars. The best T0 lines showing the lowest levels of reducing sugars were selected. Seeds were collected from these best T0 lines. T1 progeny were analyzed by qPCR to determine the effectiveness of NtSUS silencing events in reducing reducing sugars.

Manipulation of NtSUS genes (e.g., either using a constitutive promoter or a specific senescence-inducible promoter such as SAG12 or E4) can alter the chemistry of cured tobacco leaves. Similarly, knockout of NtSUS genes using a genome-editing strategy such as CRISPR or mutant selection can alter the amino acid chemistry of leaves of major commercial tobacco varieties.

Example 9. Obtaining liquid tobacco extract from tobacco plant modified NtINV4 , and tobacco plants modified NtSUS , each of which is characterized by a modulated content of reducing sugars.

The tobacco starting material is obtained from dried leaves of a tobacco plant modified with NtINV4 or a tobacco plant modified with NtSUS, according to the present invention. The tobacco material is cut to form tobacco pieces of approximately 2.5 millimeters by approximately 2.5 millimeters and the tobacco pieces are loaded into an extraction chamber without compression. The tobacco parent material is heated in the extraction chamber. During the heating, a stream of nitrogen is passed through the extraction chamber at a flow rate of approximately 40 liters per minute. For each tobacco parent material, the volatile compounds released during the heating step are collected by absorption in a liquid solvent formed from propylene glycol at a temperature of minus 10 degrees Celsius and with stirring at a speed of 750 rpm. The propylene glycol solution with the collected volatile compounds is dried in a dehydration process in order to reduce the moisture content of the solution to approximately 15 percent. Concentrated solutions of the collected volatiles from the tobacco parent materials are collected.

A combined liquid tobacco extract can be obtained. For each of the tobacco raw materials processed as described above, the first tobacco raw material is heated at a temperature and for a period of time that differ from those for the second tobacco raw material. For each tobacco raw material, volatile compounds released during the heating step are collected and dried. The resulting concentrated solutions of the collected volatile substances from the first and second tobacco raw materials can be combined in a certain ratio to obtain a liquid tobacco extract.

Any publication cited or described in this document provides relevant information disclosed prior to the filing date of this application. Statements made herein should not be construed as an admission that the inventors have no basis to contrast the present invention with earlier such disclosures. All publications mentioned in the foregoing description are hereby incorporated by reference. Various modifications and variations of the present invention will be apparent to those skilled in the art without departing from the scope and spirit of the present invention. Although the present invention has been described in connection with specific preferred embodiments, it is to be understood that the present invention, as claimed, should not be unduly limited to such specific embodiments. Indeed, various modifications of the described modes for carrying out the invention that are apparent to those skilled in the cellular, molecular, and plant biology or related fields are intended to fall within the scope of the appended claims.

TABLE 1

Expression of NtINV3 and NtINV4 genes in root, stem, midstem leaf, immature flower, sepal, and petal of field-grown Virginia tobacco plants (RNAseq, FPKM)

TABLE 2

Expression of NtINV3-S, NtINV3-T, NtINV4-S and NtINV4-T during air-curing of dark tobacco shown in Figure 2

TABLE 3

NtSUS gene expression during early curing stages for Burley (BU), Virginia (FC) and Oriental (OR) tobacco

B.U. FC OR Greens Mature 48 hours drying Greens Mature 48 hours drying Greens Mature 48 hours drying NtSUS1-S 3.4 0.2 0,0 10.4 0,0 0.4 3.4 0,1 0.5 NtSUS1-T 4.1 0.2 0,0 11.4 0,1 0.2 3.4 0.2 0.6 NtSUS2-S 0.2 0.3 28.5 0.3 0,1 1.7 0.6 2,3 17.8 NtSUS2-T 4.0 1.7 5.0 2.6 0.3 0.5 2.7 2.8 3.7 NtSUS3-S 23.4 65.6 130.4 14.2 57.8 82.3 18.7 42.0 163.9 NtSUS3-T 8.5 12.2 76.5 2.4 3.2 41.2 6.1 10.7 126.7 NtSUS4-S 11.6 4.9 22.8 15.5 3.6 22.9 19.0 18.5 105.4 NtSUS4-T 3.2 2.7 8.1 2.7 3.1 1.9 38.0 56.2 37.9 NtSUS5-S 0.6 0.2 0,1 5.1 0,1 0.5 1.4 0.3 0.8 NtSU55-T 0.5 0.5 0.3 3.6 0.4 0.6 1,1 0.8 1.6 NtSUS6-S 7.8 9.5 8.2 7.6 7.7 5.4 7.8 8.2 8.3 NtSUS6-T 3.1 4.2 4.7 1.6 4.3 3.2 3.7 4.9 5.0

TABLE 4

Expression of NtSUS genes in root, stem, midstem leaf, immature flower (immature flower), sepal and petal of field-grown Burley (BU) and Virginia (FC) tobacco plants

B.U. FC Root Stem Leaf from the middle part of the stem Invisible flower Sepal Petal Root Stem Leaf from the middle part of the stem Invisible flower Sepal Petal NtSUSl-S 206.4 294.0 0.9 8.2 1.6 6.4 95.7 129.1 0.7 31.7 1,1 0.7 NtSUSl-T 170,0 348.6 1.0 14.4 2.5 9.0 70.9 137.6 1,1 36.9 1.7 1.7 N15US2-S 26.4 17.0 0.4 86.1 12.9 97.9 27.3 13.5 0.4 197.3 7.7 150.3 NISUS2-T 61.5 55.1 2,2 96.9 37.3 172.7 4.2 12.7 0.4 122.6 12.4 111.4 NtSUS3-S 7.7 23.0 31.8 10.3 22.5 32.4 81.0 98.7 40.4 14.6 67.5 34.6 NISUS3-T 6.7 6.4 5.0 8.8 7.9 13.7 13.9 20.2 2.8 4.7 6.8 17.7 NISUS4-S 26.5 40.0 8.5 19.4 14.7 23.4 30.6 43.4 6.6 31.4 10.3 34.1 NtSUS4-T 40.7 29.3 6.4 3.2 6.0 4.1 52.6 47.7 4.8 5.4 12.6 20.4 NtSUS5-S 2.6 6.5 0.5 5.6 1.4 1,1 2,2 2,2 0.3 4.4 0.5 0.4 NtSUS5-T 2.6 5.9 0.6 5.3 1.0 1.9 2.6 2,3 1,2 3.4 0.8 1.8 NtSUS6-S 8.3 7.8 7.8 8.9 6.8 7.1 9.4 8.5 6.6 9.5 7.3 8.6 NtSUS6-T 2.7 2.4 4.1 3.0 3.1 4.9 4.0 3.4 3.6 4.2 3.2 5.2

SEQUENCE LIST

SEQ ID NO 1 : polynucleotide sequence of NtINV3-S

atggcggaaacaaacaatagcgttccttacacccaattaccggcggaggacaataacacctccgttaattctccggccggatgccggctacgacccaaaagagtgtcgtttatagtattaacagggctggtg gcagctttgttactttttgtggcagtgaaatatgggaaaaacgaggcggaggatgtgaatccagggccagtaccaccacaagaaaccgtgtgcaatatgcttggttctaatctaatgccgctgaccagcatg aagacggtggcgcgtggggtggcagaaggtgtctccgccaagtcacgcggtcgtttcttgggattacggccgtttccatggaccaaacaaatgttggcttggcaaagaacatccttccactttcaacctaag aagaattggatgaatggttagtaattctttttctcttatgttattaattttcataaatcaactttattattattattatacaataaatcaacattgcttattgatgaattttaacataaacccgccttatgct tgacgagattaactagaactatatatacaatgaatgattatctccattccattacataaccatgaattatgtttcttaattaattaaagatttgacatgacattatatttcgtttatagtttaagaaaagct ttgtattgatgtaaaagaaaccattacagcttcgaatatgggataccttgtctttttcttttcctaagatggatctttgattgcaagaacagagtttgaattactcaggaaacttatttgcttatttattatt ttttgaggtgaacattaatgatttattcttatttggcatgtgttggattatttggcttggattgcgctgatcacggaaattgcctgattcttttcgtcagatcctaatggtaaagtccatatatttctactt gttattgttgttgttcttcttattattatattattattgaaaattatcgacataatcgggacctcaaaacatactagtcgtagcagtttttaagtagacagatgtcaatatgatgaagacagttgttttcag acaattgcatgtgaattttctaggagcaaacacaaattcctagaatggtaagcaacttccaccctgtctgttccaattataacctcgctacttttgatccacttaatcttattcaaccaacagtggatcact tatttaattatatgtgacctagtttattgagacatttttacattaagccctttcgtatttacacttcaatatgcatcatacaaaaaaaaatgtacttcaaagttatacgttatattaatttctaactccaat ttttaaaaaaaaatatattttaggtcccttattctacaaaggatggtaccatttgttctatcaatacaatccagaggctgcagtatggggaaatattgtatggggacatgcagtttcaagagacttaattca ctggcaacaccttccagttgctatggttgcggatcaatggtacgacattaatggtgtatggaccggatccgcaaccattttacccgatggtaaactcgtcatgttgtatactgggtcaaccaacgagtcagta caggttcaaaatttagcgtacccggctgacccatcggatcctctcctaataaaatgggtcaagtatgagggcaacccggttcttgtaccaccacccggaattgctgctaaggatttccgtgaccccaccact gcatggaccacaccacaaggcaaatggcggattactattggttcaaaagttaataaaactggaatttcattggtctatgacactattgattttaagaattttgagttgctggatggggtgctccatggtgtat cgggtacgggtatgtgggaatgtgtggatttttacccggtttcgaaagttgttgaaaatgggcttgacacttcagataatgggcctgcagtaaaacatgtgttaaagtccagtcttgatgatgatagaaatg attattatgcacttggaacttatgatgctgtggctggaaaatgggttcctgataatcccactattgatgttggtattggattaagatatgattatggaaatttttatgcatcaaaaacattttatgaccaaga gaaaaagagaagagtcctttgggcttggattactgaaagtgatagtgaagctgctgatatttgcaaaggttgggcatcacttcaggtacaattcaattgtgtcaagctagcgcttgcacatagatttagttg aaacctaaaaaatgagtatttgaaattccgtagaaaaataatttttgaaagttgaagttgtgtttgaatatgcattttatttgaaaaaaaaaacagttctaattttatgagaaagaaaaattcacctaaaaa ctgccctaaaccagattttaggaacttgaaaaaaaaataaactttttcaaaaactgattatattcttgaacaaacaatattatcaaaaatctattttttttttgccaaaatctatggccaaacaggagcta atttcctttattttttttttcaaacttcatgtcatatttgaattttggtctcatttaacactttggtaacgtgtgatgtaacacagcccattccaaggactataaaatatgacaagaagacaggaagcaatat aattacttggccagtggcagaggttgagaatttgagatttaacagcaaggaattcgacaaggtggaggtcaagccaggaaatgttgttccactagaagttggcactgccactcaggtttgttcattaaattt agcttatatacactgactgcctaaaagaatttttttgacattattagtgtattttaagctattatagcacgtaacatgctaatgctcgaataagtttaacttactataacttgaattgttgatgattacagtt ggacataatggctgagtttgaagtagaccctaaggtcttggagaaattagaaggaagtaatgctacatatgagtgcagaagcagcggtggatctgctgaacgtggtgccttaggaccatttggtttattggt tttaacagataagggattgtccgagcaaactccaatttacttctacattgctaaagacgctgctggaaatttcaccacattcttctgcaatgatcttaccaggttctaatttctcctctcttgcattttcatc tcatcaatgaagttttagcccttcaccccctcccccaaaaccaaactaataaattggagaaaaccctttattggttcagtgcttaatagcagtacggaattcaggattttaagtcagtgggttctgcgatct atatatatataataatatttttctgcacatacatatagtccgagctagacatagtgagttccgttgaacctgttgcatttagtctgagtccgccactgcttaagcacatccttctcataacaaccgagctttc caaaaacttaagtatttctcatgtccatacttttattcatgtttgaaaatgaagtcacattttgttttataaccgaaaaatcccgagggcaagtggccagtacatggttcgaagctcaatggacactggcac cgcccctttatcgtgctccacttaaatactaagattttgtccgtggcagggtttcaaccaatcacgtacgtttaactcatatattaggaatagcttttaccactagaccaaaactcggggacaatgtatgaag ccggatatttgttgcaattctttttaaattaaaatggggacaagatccgagacaaatcttgaaaatgcattacgaagtattgttaagtaagtatgaaaatggtgattctcatcttttttacttcctttttttag gtcatctgaagcaacagatgttcgcaaactaatctacggaagcacagttccagtcctccaaggagagaagctttctctaagaacactggtaatatcccctttttctttcttaatttcttaatccaaattctta attagtgcttgttttcctttgtgcgtataattaagtttactaagtatcaattaatggggtatttttgtcaatgtaataggtggatcattcaatagtagaaagttttgcacaaaatggaaggacagcaataac atcaaggttatatccaacaaaggcaatatatgaagatgctaagctctacttgtttaacaatgctacagatgttaccattactgcctcggtcaagatttggcaaatacattctgcaaatatacaatctagttaa

SEQ ID NO 2 : polypeptide sequence of NtINV3-S

MAETNNNSVPYTQLPAEDNNTSVNSPAGCRLRPKRVSFIVLTGLVAALLLFVAVKYGKNEAEDVNPGPVPPQETVCNMLGSNLMPLTSMKTVARGVAEGVSAKSRGRFLGLRPFPWTKQMLAWQRTSFHFQPKKNWMNDPNGPLFYKGWYHLFYQYNPEAAVW GNIVWGHAVSRDLIHWQHLPVAMVADQWYDINGVWTGSATILPDGKLVMLYTGSTNESVQVQNLAYPADPSDPLLIKWVKYEGNPVLVPPPGIAAKDFRDPTTAWTTPQGKWRITIGSKVNKTGISLVYDTIDFKNFELLDGVLHGVSGTGMWECVDFYPVS KVVENGLDTSDNGPAVKHVLKSSLDDDRNDYYALGTYDAVAGKWVPDNPTIDVGIGLRYDYGNFYASKTFYDQEKKRRVLWAWITESDSEAADICKGWASLQPIPRTIKYDKKTGSNIITWPVAEVENLRFNSKEFDKVEVKPGNVVPLEVGTATQLDIMAE FEVDPKVLEKLEGSNATYECRSSGGSAERGALGPFGLLVLTDKGLSEQTPIYFYIAKDAAGNFTTFFCNDLTRSSEATDVRKLIYGSTVPVLQGEKLSLRTLVDHSIVESFAQNGRTAITSRLYPTKAIYEDAKLYLFNNATDVTITASVKIWQIHSANIQSS

SEQ ID NO 3 : polynucleotide sequence of NtINV3-T

SEQ ID NO 4 : NtINV3-T polypeptide sequence

MAETNNNSVPYTQLPAEDNNTSSNSPAKCRRRPKRVSFIVLTGLVAALLLFVAVKYGNNEAEDVNPGPVPPQETVCNMLGSNLMPLTTMRTVARGVAEGVSAKSRGRFLGLRPFPWTKQMLAWQRTSFHFQPKKNWMNGPLFYKGWYHLFYQYNPEAAVWGN IVWGHAVSRDLIHWQHLPVAMVADQWYDINGVWTGSATILPDGKLVMLYTGSTNESVQVQNLAYPADPSDPLLRKWVKYEGNPVLVPPPGIATKDFRDPTTAWTTPQGKWRITIGSKVNKTGISLVYDTIDFKKFELLDGVLHGVPGTGMWECVDFYPVSK VVENGLDTSDNGPAVKHVLKSSLDDDRNDYYALGTYDAVAGKWIPDNPTIDVGIGLRYDYGNFYASKTFYDQEKKRRVLWAWITEGDSEAADICKGWASLQPIPRTIKYDKKTGSNIITWPVAEVENLRLNSKEFDKVEVKPGSVFPLEVGTATQLDIMAE FEIDPKVLERLEGNNATYECRSSGGSAERGALGPFGLLVLTDKGLSEQTPIYFYIAKDAAGNFTTFFCNDLTRSSEATDVRKLIYGSTVPVLQGEKLSLRTLVDHSIVESFAQSGRTAITSRVYPTKAIYEDAKLYLFNNATDVSTASLKIWQMNSANIQSS

SEQ ID NO 5 : polynucleotide sequence of NtINV4-S

atggccacccaccattccattatgacccggaaaactccacgacccattacactgtcctaccggatcaacccgaatccgccggcgccgggcgccggaagtctcttaaagttgtctccggcattttgctc tcctctttctttttgctttctttagtctttgtgatcctcaaccagtcttcagatttatcacaagaaaactcccgctcgtcggagactttgacgccggcgttgtcacgaggtgtatctcagggagtttccg agaagactttcaaggatgtttccggtagaagcctttcgtactacccgtggactaatgctatgcttacttggcaaaggactgcttaccattttcaacctcaaaagaattggatgaacggtaaattttttgg cttatctttctcttattaattcttttaataaaacatgaattttaagatacttatactggctttttcttattgattcttatggctattttgttggggtatcctatggattctgattggatgatatgctgca gatcctaatggtgagtttacttattaccataattactttttattatttattattcccaaaccatgattagtgcatccggctattggttaaagattcacaaaaccaataaaatagtaatcttgtcatagt ttccataataatctacacgtacgctattgtttaatgacaagaaatttgacgctcagcatagttaattctctcatatttgtattgtttactttatagcctttgagctaattaattctgggtttctttgaac taaaccttttataagttacaatcacacatagatgagttggcacattattcaggctaataatgaaagaaattggattacttgactaatatggcaaatgcggccaatttaatttggattaacacgatatatgt gtggtaataatgcttttgtgcaacatctctcatacaaggacacatgattaggtgattttgtaccaagtctcgggaccaatcacaatatatgggtcacaccttatatattattgtaagagttgggacccac caaggatttgtctgtctttccaactagccacttgtctttttctcttttttatatttttaaatgaaatggtgtgggttttttatttttgggtcgatctaaccgcttctgcctattatcaatttagccttg tgattgtgagaatagaagagagaaatagaggataataataataaggataagaattaagaacgtaccttcttattgtcgaaattatttgagaagactattcattgttctgattagtgtccatcgatgtccc tttcctcctttttctatcttggagaggtttcctcttctttgttttacttttcctttttctaaatatgcattccaaaatcttaacactactcgaacgtccattcttggaaagtctcttttgaaagtttaggg caacatcattcggacaacttaattagcattcactattaaaaattaatagaacagaaaagttcatgtatttttttagggagagtaagaggcggattcagaatttaaatcttatgtgtttagtttttaaaat ttttaggattgataactgaacatggcgagaaacatgaacatgtgacataaattccgtgtttcaacactaaacaggtccattataccacaaaggatggtaccatcttttttatcaatacaatcctgattca gctgtttggggaaatatcacatggggccatgcaatatccacggacttgatccactggctttacttgcctttcgccatggttccgatcaatggtacgatatcaacggtgtctggaccgggtccgcgaccat cttgcccgacggtcagatcatgatgctatacaccggtgataccaatgattacgtgcaggtgcaaaatcttgcataccctgctaacttatcggatcctctctcctcatcgactgggtcaagtaccaggacaat ccggtcatggttcccccacccggcattggtgtcaaggacttcagagacccgacaactgcttggaccggaccccaaaacgggcagtggctgctaaccatcgggtccaagattggtaaaacgggtattgcac ttgtttatgatacgtccaacttcacaaactttaagctattggatggagttttgcatgcggttccgggtacgggtatgtgggagtgtgtggacttttacccggtatcaaccgttgaggcaaacgggttgg acacatcatataacgggccaggtataagcatgtgttaaaagcaagtttagatgacgataagcatgattactatgctattgggacatatgacccggtaaagaacaaatggactcctgataacccggaattg gatgtgggtatcgggttgagactggactacgggaaatactatgcgtcaaagacattttatgacccgaaagaacaaagaagaatattgtggggatggattggagaaactgacagtgaagctgctgatctgc tgaagggatgggcatctgtacaggtatggactcttttaagtacactacctcagcatccgaagagcattacacttttatttttgttttacattagaccacatgaatgggtgttttggcataactggtaaag ttgttgccatgtgaccctgaggtcacggggttcgagccgtagaaatagcctcttgcagtaatgtaataaactcttagtgcatagggttgccttttttattagaccacacacatgttcaagttatgtcatgt tagtcgtgtcaattttttgtggaaatcaatttactgcacctcaatcttgaattagttgagactagctataggaacctttgtattgagaggacttatcataatttgatcatttttgcactaactgtcacac tatgatattcactttctttatccagtttagtagtgtgccaatacaccttaagcacgtgacaagaatttattagcagggtcatctcgattttatgtaggagtacagaattgaattgaatcttttcttctag taaattctcaattgcaacttgacaatgaagtttttcagatgcaaaaaagatgaaatatctctaataatttccttttccaataacagagtattccaaggactctgctttatgacaaggagacaaggacaca tgtacttcagtggccagttaaagaaattgagagcttaagaattggtgatcctctagtgaaacaggtcaatcttcaaccaggctcaattgagcttgtccatgttgactcagccgcacaggtttgctttct catccttcgaaattgaaaacgtttcacttatatgtgcttgatgtacagtcctaaaacttgtatgcgcaatggtgcagttggatgtagaagcctcatttgaagtggacaaagcagcactcgcgggaacaat tgaagcagatgtggtttcaactgcagtactagtggaggtgctgctaaaagaggcattttgggaccatttggtgtcgttgtaattgctgatcaaacgctttctgagctaaccccagtttacttctacattg ccaaaggaactggtggccgagctgaaacctacttctgcgctgatgaaactaggtttgcttctactatgtttatcttgtatactctatcttaatagtccttgtcaaagtatagaggaataacatagcggcg tgatctgatgcagatcctcagaggctcctggagttgctaaacaagtgtatggtagttcagtaccagtgttagatggtgaacaacactcaatgagattattggtaagtgataatccctttattctgacttt cttcaaatcaagaataatatcaagcttattagttcttccagtcatcttacttaatttgtggaaatgctccaaagtagtcaatttggtaactattcaagataatgtggttcagaataatttgtgttatgaa tgtatttgacagttgggatgatctgttttttagtaaaatttcttaaaaacttaattcaggtggaccactcaattgtggaaagctttgctcaaggaggaagaacagtcataacatcgcgaatttacccaac aaaagcaatcaatggagcagcacgactgttcgttttcaacaatgccaccggggctagtgtgactgcctccctcaagatttggtcactcaaatcagctgatattcgatccttccccttggaccagttgtaa

SEQ ID NO 6 : polypeptide sequence of NtINV4-S

MATHHSHYDPENSTTHYTVLPDQPESAGAGRRKSLKVVSGILLSSFFLLSLVFVILNQSSDLSQENSRSSETLTPALSRGVSQGVSEKTFKDVSGRSLSYYPWTNAMLTWQRTAYHFQPQKNWMNDPNGPLYHKGWYHLFYQYNPDSAVWGNITWGHAIS TDLIHWLYLPFAMVPDQWYDINGVWTGSATILPDGQIMMLYTGDTNDYVQVQNLAYPANLSDPLLIDWVKYQDNPVMVPPPGIGVKDFRDPTTAWTGPQNGQWLLTIGSKIGKTGIALVYDTSNFTNFKLLDGVLHAVPGTGMWECVDFYPVSTVEANGLD TSYNGPGIKHVLKASLDDDKHDYYAIGTYDPVKNKWTPDNPELDVGIGLRLDYGKYYASKTFYDPKEQRRILWGWIGETDSEAADLLKGWASVQSIPRTLLYDKETRTHVLQWPVKEIESLRIGDPLVKQVNLQPGSIELVHVDSAAQLDVEASFEVDKA ALAGTIEADVGFNCSTSGGAAKRGILGPFGVVVIADQTLSELTPVYFYIAKGTGGRAETYFCADETRSSEAPGVAKQVYGSSVPVLDGEQHSMRLLVDHSIVESFAQGGRTVITSRIYPTKAINGAARFVFNNATGASVTASLKIWSLKSADIRSFPLDQL

SEQ ID NO 7 : polynucleotide sequence of NtINV4-T

atgatgttatacaccggtgataccaatgattacgtgcaggtgcaaaatcttgcgtaccccgccaacttatcggatcccctcctcatcgactgggtcaagtaccggggcaacccggtcat ggttccaccacccggcattggtgtcaaggactttagagacccaacgactgcttggaccggaccacaaaacgggcagtggctgcttaccatcgggtccaagattggtaaaacgggtattgc aattgtttatggtacttccaacttcacaaactttaagctattggatggagttttgcatgcggttccgggtacgggtatgtgggagtgtgtggacttttacccggtatcaaccgatgaggc aaacgggttggacacatcatataacgggccaggtataaagcatgtgttaaaagcaagtttagatgacgataagcatgattactatgctattgggacatatgaccggtaaagaacaaatgg actcctgataacccgcaattggatgtgggtatcgggttgagactggactacgggaaatactatgcgtcaaagacattttatgacccgaaggaacaaagaagaatattgtggggatggatt ggggaaactgacagtgaagctgctgatctgctgaagggatgggcatctgtacaggtatggacacttttcaagtacactacctcagcttccgaagagcattacacatttatttttgtatta cattagggtgccttggcgtaactctggtaaagtaagttctgaattgcaacgtgaaaatggaggtttttagatgcaaagagatgatatatccctaatagttttcctgttttaataacagag tattccaaggactgtgctttatgataaggagactaggacacatgttcttcagtggccagttaaagaaattgagagcttaagaattggtgatcctctagtgaaacgggtcaatcttcaacc aggctcaattgagctagtccatgttgactcagccgcacaggttgctttctcatccttggaaattgaaaacgtttcacttatatgtgcttaatgtgcagtcctaaaacttgtatgtgcaat ggtgcagttggatgtagaagcctcatttgaagtggacaaagcagcactcgagggaacaattgaagcagatgttggtttcaactgcagtactagtggaggtgctgctaaaagaggcatttt gggaccatttggtgtcgttgtaattgctgatcaaacgctttctgagctaactccagtttacttctacattgccaaaggacctgatggccgagctgaaacctacttctgtgctgatgaaac taggtttgcttctactatgtttatcttgtatactctatcttaatagtccttgtcaaagtatagatgaataacatagcggcgtgatctgatgcagatcctcagaggctcctggagttgcta aacaagtgtatggtagttcagtaccagtgttagatgatgaacaacactcaatgagattattggtaagtgataatcccgttattctgaccttcgtcaaatcagaataatatcaagcttatt agttcttccagtcatcttattaaatttatggaaatgctccaaagtagtcaatttggtaactattcaagataatgtggttcagaataatttgtgttatgaatgtatttgacagttgggatg atctgtgtttttgagtaaaatttcttaaaactgaactcaggtggaccactcaattgtggagagctttgctcaagggaggaagaacagtcataacatcgcgaatttacccaacaaaggcaat caatggagcagcacgactgttcgttttcaacaatgccacgagggcaaggtgactgcctccctgaagatttggtcactcgaatcagctgatattcgatccttccccttggaccagttgtaa

SEQ ID NO 8 : polypeptide sequence of NtINV4-T

MATHHSHYDPENSTTHYTVLPDQPESAGSGHRKSLKVVSGILLSSFFLLSLVFVIVNQSSDLSQKNSHSSETLTPALSRGVSQGVSEKTFRDVSGGLSSYYPWTNAMLTWQRTAYHFQPQKNWMNGPLYHKGWYHLFYQYNPDSAIWGNITWGHAISTD LIHWLYLPFALVPDQWYDINGVWTGSATFLPDGQIMMLYTGDTNDYVQVQNLAYPANLSDPLLIDWVKYRGNPVMVPPPGIGVKDFRDPTTAWTGPQNGQWLLTIGSKIGKTGIAIVYGTSNFTNFKLLDGVLHAVPGTGMWECVDFYPVSTDEANGLDT SYNGPGIKHVLKASLDDDKHDYYAIGTYDPVKNKWTPDNPQLDVGIGLRLDYGKYYASKTFYDPKEQRRILWGWIGETDSEAADLLKGWASVQSIPRTVLYDKETRTHVLQWPVKEIESLRIGDPLVKRVNLQPGSIELVHVDSAAQLDVEASFEVDKA ALEGTIEADVGFNCSTSGGAAKRGILGPFGVVVIADQTLSELTPVYFYIAKGPDGRAETYFCADETRSSEAPGVAKQVYGSSVPVLDDEQHSMRLLVDHSIVESFAQGGRTVITSRIYPTKAINGAARLFVFNNATRSVTASLKIWSLESADIRSFPLDQ

SEQ ID NO 9 : Nucleotide sequence used for silencing of NtINV4-T and NtINV4-S

ggtttcaactgcagtactagtggaggtgctgctaaaagaggcattttgggaccatttggtgtcgttgtaattgctgatcaaacgctttctgagctaac

SEQ ID NO: 10 : polynucleotide sequence of NtSUS1-S

atggcagctagtggtcttagcattaagaaaagtttggaggaatccattttggctcatccagatgaaattttggctctcaagtcaaggtacattactacatataatgatattaagaactagaggcttatccaaggttt tgttacatttttgaaattataagtttagaacctaatagtacttggtagcacttgtttccttattatctagctgttgttactgcttgttgctactgctttctgttcatctttccttgagcccggtctatcggaaacaa cctctctattctcaaagtataaggtttgcgtacatactacctccccagactctacttgtggaatttactgtttttgttgtgttgttgtaatctaatatttattagaattttactgatttttcacatatatatatcta tgtcccctgtcgaaaattctatagctcatgttagctaaatacattagtaccattgtttttaattgttttggttttggcacaggattgaaactgaagggaaaggggtaatgaaaccacttgatctcttgaaccatttg gtttctgttactagtaagacaaatggagtaaatattgtacctagtgcacttgtggaagttctcagttgcagccaagaagctgtgattgtaccaccaaaactagcactagctgtacgtccgaggcccggtgtatggga gtacttgtcactgaatcttaagacaaagaaagtggctgaattaagcattcctgaataccttcaattgaaagagaacactgttgatgaaaggtaaagtattagtctgcgatttcgctttgtgaaattgaagtttttgt tttgattcataatgttttgtgtatcaattatgttaccagtggaaacatattggagttggattttgagccatttacaacagttacaccaccaaaaacactttctgactctattggcaatggtttggagtttcttaatc gccacattgcttcgaaaatgtttcatgataaggagattccagatgcctccttgacttcctcagaaaccataactacaaaggaaaggtaataaaaaaaagtgtttctttaaacaagttgtatgattatgtgtatatt tctaagtatgttaacttgaaaacagtcattgatggtgaaagaaagcattcaaagcctagagagtttccaacttgttctgaaaaaagcagaggaacatttgtgcacattgaatccagaaactccatactccaattttg aatcaaagtttgaagagattggcttggaaagagggtggggaaacaccgctgaacgcgtgcaagacactatcagtcatcttttgcatctccttgaggctcctaacgcgtcttctttggaaaatttccttggtagaatc ccattggttttcaatgttgtgattctaactccacatggttattttgctcaagataatgtcttgggctatcctgacactggtggccaggtttgtgtccaatattttgcattcttgatcaagttcttttaccatttga accaacaatcttnaacattctcttttttggttgtgaaatgttgaataggttgtttacattcttgatcaagttccagctatggagcgtgagatgcttcatcgtatgaagcttcaaggactcgatgatatcatccctcgc atccttgttgtaagtggccttaattttcctagtttcatttacacctctaaatgaaattgatcttttttgttgttttatatcaggtaacaaggctgctgcctgatgcagtaggaaccacctgtggcgagcggatggag aaagtatatggggcagaacattctcatataattcgtgttccatttagaactgagaagggaatgttgcgcaaatggatctcacgattcgaagtctggccatacatggaaactttcactgaggttggaacataaaaaca aataaaatccattggaatgttccttctgcaattgaaaatgtcttgctaactgaagacccatttttaaattgatcatcaggatgttgcagaagaacttgtcaaagaattgcaagctaaaccagacttgatcattggaa actacagtgagggaaatcttgctgcctctttgcttgcgaagaaatttggggctactcagtgtactattgctcatgccttggaaaaaactaagtatccaaactctgaccttaattggaagaagtttgatgacaagtat catttctcaagtcagttcactgctgatctctttgccatgaatcacactgatttcatcatcaccagcactttccaagaaattgctggaaggtaaaagcaaatgcacaccatcatagtatttcatatttttacccttgt ttatactatttccattcaccgaccccgacttgtttaggattgagccatagttgttgttgttgtttgtttatactatttccatttgccgaccacaacttgtttaggactgaggtatagttgttgttgttggtttgttc atattattttcattcgctaaccctaacttgtttgggactgaggcatagtagtagtagtagttgttgctattagtttatactatttccatttgccaaccccaacttgtttggtactgagacatagttgttgttgttgt tgtttgtttatactatttccatttgccgaccccaacttgtttaggactgaggtatagttgttgttgttggtttgttcatattattttcattcgctaaccccaacttgtttgggactgaggcatagtagtagtag tagttgttgctattagtttatactatttccattgccaaccccaacttgtttggtactgagacatagttgttgttgttgtttgtttatactatttcaatttgtcgaccccaatttgtttgggaccaaggcatggttg ttgttgttgtttgtttgtttttactgtttccattgatattggaacatttgttatttgcagcaaaaacactgtaggacagtatgagagtcatactgctttttaccatgcctggattgtaccgagtagtccatggaatcg attcgtttgatccaaagttcaacattgtctcccctggggctgatatgtcaatctacttcccttacactgagaaggagaaaaggctaaccaacttccacccggaaattgaagaactcctctacagtcctgttgagaat aaggaccacttgttagtctccttaatttgcttttatttcatcccatttatgatcgcttttatcccaacagatcgattaatcatttgttatcaacataaacagatgtgtgttgaaggaccggaacaagccaattctct ttaccatggcaaggctagatcgcgtgaagaatctaacagggctcgtggaatggtatgctaagaatgcaaggctgagggagcttgttaaccttgtggttgtaggcggagacagaaggaaagaatccaaagatttagaa gagcaagcagagatgaagaagatgtatgatcttatcgaaacctataacctgaacggccaattcaggtggatttcttcccaaatgaatcgtgtgaggaacggagaactctatcgttacattgcagacacgaggggtgc tttcgttcaaccagcattctacgaggcttttggtttgacagttgtagagtctatgacttgtggtttgccaacttttgctacttgtaatggtggaccatttgagattatagtgaatggaaaatctggtttccatattg atcctaatcaaggtgacaaggctgctgatatgttggtaaatttctttgaaaaatctaaagaagatccaagttattgggatgctatttccaagggaggtctgcaacgtattcttgaaaagtaagcttttgcatttgat tagcacaagtgcacaaccaagatttaacttttgaacaaactaaaactaacccttttttgtattttctctttgctaggtatacatggcaaatttattcacagaaagtgatcacactatctgggatttatggattctgga agtatgcaaccaagaatgataaagttgctagtgcaaagaagcgctatcttgagatgttttatgaacttggatttaagaaatcagtaagtgtcaattttaaaggggaaccttggatcaacggttaagttgtctttgtg caacctataggtcaggggtttgagccgtagaagtagccactaatatttacattagggtagactgtgtacatatcacaccccttggggtacggccctttcctggatcctgtatgaacgcgggatgccttgtgcaccgg gctgtatttttttttttagtgtcacttctgtattttgtttgagcttgtttataaagtttggaaatctgctgctaatttgtatatttgttggttgtgtatttcaggctgagaaagttccattggctattgatgaatag

SEQ ID NO: 11 : NtSUS1-S polypeptide sequence

MAASGLSIKKSLEESILAHPDEILALKSRIETEGKGVMKPLDLLNHLVSVTSKTNGVNIVPSALVEVLSCSQEAVIVPPKLALAVRPRPGVWEYLSLNLK TKKVAELSIPEYLQLKENTVDESGNILELDFEPFTTVTPPKTLSDSIGNGLEFLNRHIASKMFHDKEISRCLLDFLRNHNYKGKSLMVKESIQSLESFQL VLKKAEEHLCTLNPETPYSNFESKFEEIGLERGWGNTAERVQDTISHLLHLLEAPNASSLENFLGRIPLVFNVVILTPHGYFAQDNVLGYPDTGGQVVYI LDQVPAMEREMLHRMKLQGLDDIIPRILVVTRLLPDAVGTTCGERMEKVYGAEHSHIIRVPFRTEKGMLRKWISRFEVWPYMETFTEDVAEELVKELQAKP DLIIGNYSEGNLAASLLAKKFGATQCTIAHALEKTKYPNSDLNWKKFDDKYHFSSQFTADLFAMNHTDFIITSTFQEIAGSKNTVGQYESHTAFTMPGLY RVVHGIDSFDPKFNIVSPGADMSIYFPYTEKEKRLTNFHPEIEELLYSPVENKDHLCVLKDRNKPILFTMARLDRVKNLTGLVEWYAKNARLRELVNLVVV GGDRRKESKDLEEQAEMKKMYDLIETYNLNGQFRWISSQMNRVRNGELYRYIADTRGAFVQPAFYEAFGLTVVESMTCGLPTFATCNGGPFEIIVNGKSG FHIDPNQGDKAADMLVNFFEKSKEDPSYWDAISKGGLQRILEKYTWQIYSQKVITLSGIYGFWKYATKNDKVASAKKRYLEMFYELGFKKSAEKVPLAIDE

SEQ ID NO: 12 : polynucleotide sequence of NtSUS1-T

atggcaggcagtggtcttagcattaaggaaagtttggaggaatccattttggctcatccagatgaaattttggctctcaagtcaaggtacattactgcataatgatattaagacctagaagc ggatccaagattttgttacatttttgaaattataagtttagaatctaatatttgttatcgcttgtttccttattatcttgctgttgttactgcctgttgctactagtttctgttcatccttcc ttgagctgagtttctatcggaaacaacctctctactctcaaagtaggaataagttatgcgtacacactaccctccccagactccacttgtgtaatttactgagtttgttgttgttgttgttg taatctaatacttgttagaattttactgatttttcacatatatatctatgacccatgtcgaaaatactatagctcatgtgctaaatacattagtaccattgttttgtaattgttttggttttg gaacaggattgaaactgaagggaaaggggtaatgaaaccagttgatctcttgaaccatttggtttctgttactagtaaaacaaatggagtaaatgttgtacctagtgcacttgtggaagttc tcagttgcagccaagaagctgtgattgtaccaccaaaactagcactagctgtacgtccgaggcccggtgtatgggagtacttgtcactgaatcttaagacaaagaaagtggctgaattgagca ttcctgagtaccttcaattgaaagagaatactgttgatgaaaggtaaagtaatagtctgcgatttcgctttgtgaaattgaagttttttgtttgattcttaatgttttgtgtatcaattatgt taccagtggaaacatcttggagttggattttgagccatttacaactgttacaacaccaaaaacactttctgactctattggcaatggtttggagtttcttaatcgccacattgcttcgaaaat gtttcttgataaggagatgccaagtgcctccttgactttctcagaaaccataactacaaaggaaaggtagtaaaaaaaagtgtttctttaaacaagttgtatgattatgtgtgtatttctaa atatgtcaatttgaaaacagtcattgatggtgaaagaaagcattcaaagcctggagagtttccaacttgttctgaaaaaagcagaggaatatttgcacacactgaatccagaaactccatact ccaaatttgaatccaagtttgaagagattggcttggaaagagggtggggaaacaccgctgaacgcgtgcaagacaccattagtcatcttttgcatctccttgaggctcctaacgcgtcttcct tggaaaatttccttggtagaatcccattggttttcaatgttgtgattctcaccccacatggttattttgctcaagataatgtcttgggctatcctgacactggtggccaggtttgtgtccgat ataacatatcaagaaattttgcattcttgatcatgttctttataccatttgaaccaacattctttttttggttgtgaaatgttgaataggttgtttacattcttgatcaagttccagctatg gagcgtgagatgcttcatcgtatgaagcttcaaggactcgacgatatcatccctcgcatccttgttgtaagtgcccttaattttcctggtttggtttacctctaaatgaaattgattttctgg ctttctaacttttttggattgatctttttgttgttttatatcaggtaactaggctgctgcctgatgctgtaggaaccacttgtggcgagtggatggagaaagtatatggggcagaacattctc atataattcgtgttccattagaactgagaaaggaatgttgcgcaaatggatctcacgattcgaagtctggccatacatggaaactttcactgaggttggaacataaaaacaaataaaaatca ttggaatgttcttctgcatttgaaaatgtcttgctaactaaagactcatttttaaattaatcatcaggatgttgcagaagaacttgtcaaagaattgcaagctaaaccagacttgataattg gaaactacagtgagggaaatcttgctgcctcattgcttgctaagaaatttggggctactcagtgtactattgctcatgccttggaaaaaactaagtatccaaactctgaccttaattggaaga agtttgatgacaagtatcatttctcaagtcagttcactgctgatctttttgccatgaatcacactgatttcattatcaccagcactttccaagaaattgctggaaggtaaaagcaaatgcac accatcatagtatttcatatttttaccctagtttatactatttccatttgtcaactccaacttgtttgggattgaaccatagttgttgtttgtttatactatttccattcgccgaccccaact tatttgggactgagacataattgttgttattattgtttgtttgtttatactatttccattctcagaccccaacttctttgggactgagccgtagattgttgttgttgttgttgttgttgttt gtttatgctatttccgttcaccgaccccaacttatttgggactgaggtgtagaagtagtcgttgttgtttgtttatacgacttccaattgatattcgaatgtttttatttttgcagcaagaac actgtaggacagtatgagagtcatactgcttttaccatgcctggattgtatcgagtagtccatggaatcaattcgtttgatccaaagttcaacattgtctcccctggggctgatatgtcaatc tacttcccttacactgagaaggagaaaagactaaccaacttccacccggaaattgaagaactcctctacagtcctgttgagaataaggaccacttgttagtcttcttttatttcattcattttt ctacaccttttttttcaacagattgattgattggttcttatcaacgtaaacagatgtgtgttgaaggaccagaacaagccaattctctttaccatggcaaggctagatcgcgtgaagaatct aacagggctcgtggaatggtatgcaaagaatgcaaggctaagggagctcgttaaccttgtggttgtaggcggagacagaaggaaagaatccaaagatttagaagagcaagcagagatgaagaa gatgtatgatcttatcgaaacatacaacctgaatggccaattcaggtggatttcttcccaaatgaatcgtgtgaggaacggagaactttatcgatacattgcagacacgaggggtgctttcgt tcaaccagcattttatgaggcatttggtttgacagttgttgagtctatgacttgtggtttgccaacttttgctacttgtaatggtggaccatttgagattatagtgaatggaaaatctggttt ccatattgatcctaatcaaggtgacaaggctgctgatatgttggttaatttcttcgaaaaatctaaagaagatccaagttattgggatatttccaagggtggtctgcagcgtattcttg aaaagtaagcttttgcatttgattagcacaagtgtacaaccaagatttaacttatgaacaaactaaaactaaccctttttttattttcttttgctaggtatacatggcaaatttattcacaga aagtgatcacattatctgggatttatggattctggaaatatgcaaccaagaatgacaaagttgctagtgcgaagaagcgctatcttgaaatgttttatgaatttgggtttaagaaatcagtaa gtgtcacttctgtattttgtttgagcttgtttgtaaagtttggcaatcttctgctaatttgtactatatttgttgacttgtgcatttcaggctgagaaagttccattggctattgatgaatag

SEQ ID NO: 13 : NtSUS1-T polypeptide sequence

MAGSGLSIKESLEESILAHPDEILALKSRIETEGKGVMKPVDLLNHLVSVTSKTNGVNVVPSALVEVLSCSQEAVIVPPKLALAVRPRPGVWEYLSLNLK TKKVAELSIPEYLQLKENTVDESGNILELDFEPFTTVTTPKTLSDSIGNGLEFLNRHIASKMFLDKEIAKCLLDFLRNHNYKGKSLMVKESIQSLESFQL VLKKAEEYLHTLNPETPYSKFESKFEEIGLERGWGNTAERVQDTISHLLHLLEAPNASSLENFLGRIPLVFNVVILTPHGYFAQDNVLGYPDTGGQVVYI LDQVPAMEREMLHRMKLQGLDDIIPRILVVTRLLPDAVGTTCGEWMEKVYGAEHSHIIRVPFRTEKGMLRKWISRFEVWPYMETFTEDVAEELVKELQAKP DLIIGNYSEGNLAASLLAKKFGATQCTIAHALEKTKYPNSDLNWKKFDDKYHFSSQFTADLFAMNHTDFIITSTFQEIAGSKNTVGQYESHTAFTMPGLY RVVHGINSFDPKFNIVSPGADMSIYFPYTEKEKRLTNFHPEIEELLYSPVENKDHLCVLKDQNKPILFTMARLDRVKNLTGLVEWYAKNARLRELVNLVVV GGDRRKESKDLEEQAEMKKMYDLIETYNLNGQFRWISSQMNRVRNGELYRYIADTRGAFVQPAFYEAFGLTVVESMTCGLPTFATCNGGPFEIIVNGKSG FHIDPNQGDKAADMLVNFFEKSKEDPSYWDTISKGGLQRILEKYTWQIYSQKVITLSGIYGFWKYATKNDKVASAKKRYLEMFYEFGFKKSAEKVPLAIDE

SEQ ID NO: 14 : polynucleotide sequence of NtSUS2-S

atggctgaacgtgctctgactcgtgttcacagccttcgtgaacgtcttgatgccactttggctgcacatcgcaatgagatattgctgtttctttcaaggtattgcctaagtagtgttcttgtttcctacaaaagattcagt tggtgttcaaaaaacgatatgtgatttgatttatctgcctaagtcttggtagtcataattatccggtacctgtgctggtgcgagttagctggttcggaaactactctcttatgaaaacgagagatttagttggtgttgtctgca attctgtagtatggactattaagcagatagatcatgtttgatatcgaaaaggaatgtatatgtgatgttacttgaactggttttggttattacaggattgaaagccatggaaaagggatcttgaaacctcaccagctattg gctgagttcgatgcaattcgccaagatgacaaaaagaagctgaatgatcatgcatttgaagaactcctgaaatctactcaggtaattttgattttggctaaatgtgttaccaagctgaatgatcatgcatttgagtttgtgt ccgactactacaatgatatgttataccaggaagcgattgttctgccaccttgggttgcacttgccattcgtttgaggcctggtgtgtgggaatatgtccgtgtgaatgttaatgctctagtcgttgaggagctgaccgtcc ctgagtatttgcattttaaggaagaacttgttgatggaacgtaagttttagtctcttatttgatactatgttagagaataggcagtggattcaatttatcagtgttgttttttacctaatgcagctccaatggaaatttcgt tctcgagttggattttgagcccttcactgcatcctttcctaaaccgaccctcaccaaatctattgggaatggagttgaattcctcaataggcacctttctgcgaaaatgttccatgacaaggaaagcatgaccccgcttctt gaatttcttcgggttcacaattataagggcaaggtaactttgttattcccattcatatatatgttcagtttgtgcttatcatgcgcccaatgatgtatgaatatgtactaaaggatagatgtacgatttcgtttgcagacaa tgatgctgaatgacagaatacagaatttaaccactctgcaaaatgtcctaaggaaggcagaggaataccttattatgcttccccctgaaactccattttccgaattcgaacacaagttccaagaaattggattggagaagg gatggggcgacactgcggagcgcgtgctagagatgatatgcatgcttcttgatctacttgaggctcccgactcctgtactcttgagaagttcctagggagaattcctatggtgttcaacgtggttatcctttccccccatgg atatttcgcccaggaaaatgtcttgggttatcccgacactggtggccaggtgcattactttagtctttgtccgtgagtctatgttgctcagatcctctacaatgccactgtacccgtgtaggatactccaaatataatgca tttttggaggatctgtcaccggtgcaatggcattttggaggtcggagcaacaaacaactgctagtatgcttctaaagcttgcttccataaatgctaaggtccttcacccgtaatgtgcaggttgtctacatattagatcaag ttccagccttggagcgtgaaatgcttaaacgcctaaaggagcaaggacttgatataacaccgcgtattcttattgttagtatttcttgtacttgtaattgctgcggattacacaaaattttctctttattggcaacttatc ttgatattattcccaggttactcgtctgctgcctgatgcagttggaacaacttgtggtcagcggcttgagaaggtgtatggagccgagcactcacatattcttagggtcccctttaggaccgagaagggcattgttcgcaaa tggatatctcgctttgaagtgtggccatacatggagactttcactgaggtgacactaagcttccttgtatttgtctatcttctaattggtattaggaacaatttgctaattattaacgctttggcttttcgtacatcaggat gttgcaaaagaacttgctgcagaactgcaggccaagccagatttgataattggcaactatagcgagggaaatcttgtggcttcattgctggctcacaagttaggcgtaacgcaggtctgtgttatttttcacctcttataaa tctgattgtatttccattagtctggaactaaaagtactaaaattttcttttcttcgctgtgttatttgccttctgcagtgcaccattgcccatgcattggagaaaacaaagtatcctgattctgacatctactggaaaaaa tttgacgaaaaataccatttctcgtcccagtttaccgctgatcttattgcaatgaatcacaccgattttatcatcaccagcactttccaggagatagcaggaaggtataacatcaattgctaattcggttgcagtaacattt tgttcgatttcttccccttatgcttaacctaataccctaatgaattttccagcaaggacactgtcggacagtacgagagtcaccaggcattcacaatgcctggattgtacagagtcgttcacggcattgatgtgttcgatc ccaaattcaacattgtctcacctggagctgatataaacctgtatttcccatattccgagaaggaaaagagattgacagcacttcacccagaaattgaggagcttctgtacagtgatgttgagaacgaggaacatctgtaagt ttctaacttactcgtaccgtcagtggcagagccagaattttcattaaaatggggtcaaaatataaagacataaattcacaaagaagccaaggggtgtcaatatgtagtataaatatattaaaaaattacctagctacaca atgtaattttccgacaaaggggtatcggttgcacttcttgaatacatgtggctctgccactgggtacagttacaaagtcctgttacctatgtagatgagcttgtgctgaacatgttgtgattttggtaggtgtgtgctaaag gacaggaataagccaatcttattcacaatggcgagattggatcgtgtgaagaacttaaccggacttgttgagtggtacgccaagaacgcacggctaagggagttggttaaccttgttgtcgttggtggagaccgaaggaagg aatccaaagatttggaagagcaagcagagatgaagaagatgtatgagctaataaagactcacaacttaaatggccaattcagatggatttcttcacagatgaaccgagtaaggaacggcgaactctaccgatacattgccga cactaggggagctttcgtgcagcctgcattctatgaggctttcggtttgactgttgttgaggccatgacctgtggtttgcctacatttgcaactaatcatggcggtccagctgagatcatcgttaacggaaaatccggctt ccatatcgatccatatcacggtgagcaagctgctgatctgctagctgatttctttgagaaatgtaagacggaaccttctcattgggaaactatttcaaccggtggcctgaagcgcatccaagagaagtaagcaactctttct tgactctagtcattcaaattaacttgggatttgaggcatagttgattgataatttatcgcgtctctactactatatacaggtacacgtggcaaatctactcggagagattattgacgttggctgctgtttacggtttctgg aaacatgtttctaagcttgatcgtctagaaatccgtcgatatctagaaatgttttatgctctcaaataccggaagatggtgagttcttctgcttcctgctcttctcatagtgtttaatatacacttgattgattgcattcac ttagactaagttgctcggacacgggtgtggatgtccgacacgagtgcggatctagagttcagatccttcaagatgtaaattataagattcggggatatggatcctagtacggatacgggtgcgagaatccggctaaaaata attttaaaaaaaattatctctaaattatgagatattatgtggaatacttacgtataacttgtaaagtgtagattttttttaattctcaagttgtagattagtaaatgattgatttcctagataagtatgctattttcttcaa atttactcttctgatttcgaaaatcaaattgtatctcgtctcgaatttttccgtccgttatggtcaaagtacccaaaatcgtttgaccaaatcggtacggatcccatacccacaccacactagtgtcgtattgacacgggt gccgcacctaaactgctatgtcggagcaacttagcacttagagaatcattgatgttaaattttcttaattcttgaatctgctaatgaagattttatcttggtttttgtttaggctgaagctgttccattggctgctgaatga

SEQ ID NO: 15 : NtSUS2-S polypeptide sequence

MAERALTRVHSLRERLDATLAAHRNEILLFLSRIESHGKGILKPHQLLAEFDAIRQDDKKKLNDHAFEELLKSTQEAIVLPPWVALAIRLRPGVWEYVRV NVNALVVEELTVPEYLHFKEELVDGTSNGNFVLELDFEPFTASFPKPTLTKSIGNGVEFLNRHLSAKMFHDKESMTPLLEFLRVHNYKGKTMMLNDRIQNL TTLQNVLRKAEEYLIMLPPETPFSEFEHKFQEIGLEKGWGDTAERVLEMICMLLDLLEAPDSCTLEKFLGRIPMVFNVVILSPHGYFAQENVLGYPDTGG QVVYILDQVPALEREMLKRLKEQGLDITPRILIVTRLLPDAVGTTCGQRLEKVYGAEHSHILRVPFRTEKGIVRKWISRFEVWPYMETFTEDVAKELAAEL QAKPDLIIGNYSEGNLVASLLAHKLGVTQCTIAHALEKTKYPDSDIYWKKFDEKYHFSSQFTADLIAMNHTDFIITSTFQEIAGSKDTVGQYESHQAFTM PGLYRVVHGIDVFDPKFNIVSPGADINLYFPYSEKEKRLTALHPEIEELLYSDVENEEHLCVLKDRNKPILFTMARLDRVKNLTGLVEWYAKNARLRELVN LVVVGGDRRKESKDLEEQAEMKKMYELIKTHNLNGQFRWISSQMNRVRNGELYRYIADTRGAFVQPAFYEAFGLTVVEAMTCGLPTFATNHGGPAEIIVNG KSGFHIDPYHGEQAADLLADFFEKCKTEPSHWETISTGGLKRIQEKYTWQIYSERLLTLAAVYGFWKHVSKLDRLEIRRYLEMFYALKYRKMAEAVPLAAE

SEQ ID NO: 16 : polynucleotide sequence of NtSUS2-T

atgctttttatgggagtaaattttatggccggtcattcaactttgtgttcattacgcaaaagtcatttttcttggtgtttattacgcaagtc atttttcttttttttttgttacgtaaaaatcattcaactatgtgtttattatctaaaattcaattttttttttccttttgttacacaaaaat cattttactttactctatttatcacaaaagtcaccttggccagattttataataggcttttatcttttgttacacaaaaattattttactttactctattttatcacaaaagtcaccttggccagattttataataggcttttatcttttgttacacaaaaattattttactttactctatttat cacaaaagtcaccttggccagattttacaatacttttaccttaaaagactattatgcccttgacattataaatcctctcatttataataccttctatatgatacactatataatatatttttacctaggtattttacttataattaaaataatattaaattattttatttatctattttata atatattcatacatttaattttttcatggcaaatcactttgtttaatcatatttaaacatgaacaaattttaaatatcaaaaaaataaaaaaataaaaaaaatatttatttgaaataataacaaacagatttgtttaacaaatgatagtttttttttatagtcaataaaatttttaaaaaaattc aaagatatttgtttttaatattaatatttttaaagctttatctgttaatattatttatttgaaagtattaatctgatgtgtcattgtgttaaatgtgagtattttatttattggattaatgagtatggcttggctgataaaaagctttgattttataattttcattaaaaatattttattaagc tagtacctgacaaatttaatatcttgaaaattaacgttaagaaaaattaaatataaaaatatattataaaaataataaataaataatatcaagttattttaattataaataaaatacatggttaaaaatatattatatagcatataatatagaaggtattacataaatgagatgatttaaaggg cataatagacttttcaggtgaatgatttgtaaaatatggttaaagtgattattgtgataattagagcatagtaaaataatttttatgtaacaaaagaaaaaaaaaatgacttttgggtaatgaacataaatttgaataacttttacgtaacaaaagaataaaataaattttggataataaacat aaaattgaatgaccacctataaaatttattatttttttggggctcttcttgatttgattttttagtttagcctttgcagtaatcttggttgtcacgcgtagcgttgtgctttcgccacataagtatttagtagacttaattaatgtcattatatcggttggtgtggttttaattacttaactgtac tattatattaggtggaaggtttgaaaatttatagtagtaacattctagatcattgaaaatattggtgtttcagtgactttttagtatgtcattttcattttctaagtggttgtactaatatagtatattaaaattttgattggttgagaaacaatctctctcacctacacggtacgggtaaggt atgcgtatacgcttatcctccctacactccatttgtgggactattgttgttattttggataagctgaggtatccatcttctactaactgcactagtttattttttttgctgtttacagttgaaacaattgtctgaggatttctcacctgctgaatcaactgcaatggctgaacgtgtgctgact cgtgttcacagccttcgtgaacgtcttgatgctactttggctgctcatcgcaatgagatattactgtttctttcaaggtatagccaaagatagtattcttgttaactaaaaagattcagttggtgttcaaaaaacgatacgtttatctgcctaagtcttggtagtcagaattatccggtacct atgctggtgtgagttagctggctaggaaaccactcttatgaaaacaagagatttagttagagttgtctgtaattctgtagtatggactatgtatgtgatgctatttgaactggttttggttattataggattgaaagccatggaaaagggatcttgaaaccgcatcagctattggctgagtttga tgcaattcgccaagatgacaaaaagaaactgaatgatcatgcatttgaagaactcctgaagtccactcaggtaatatggttttggctatatttgtcgccaacgccaagctcatatttttatattattttgagcttgtgtctgaatacgacgatgatatgttatactaggaagcaattgttctgc caccttgggttgcacttgcgattcgtttgaggcctggtgtgtgggaatatgtccgtgtgaatgtcaatgcgctagtcgttgaggagctgact gtccctgagtatttgcatttcaaggaagaacttgtcgatggaacgtaagtgttagtcttcaatttgatgctatgttagagaataggctgtgga atttattgatcaatgctgtgctttgtcctgatacagctccaatggaaatttcgttctcgagttggattttgagcccttcaccgcatcctttcctaaaccaaccctcaccaaatctatcggaaatggagttgaattcctcaataggcacctctctgcgaaaatgttccatgacaaggaaagcatg accccgcttcttgaatttcttcgggttcacaattataagggcaaggtgacttgctatttccatttatctataggttcggtttgtgcttatcatgcgcccaatgacatatgaatatgcgctaaaggatatatgatttcctttgcagacaatgatgctgaacgacagaatacagaatttaacc acactgcaaaatgtcctaaggaaggcagaggaatacctcattatgcttccccctgaaactccattttccgaattcgaacacaagttccaagaaattggattggagaagggatggggcgacactgcagagcgcgtgctggagatgatatgcatgcttcttgatctcctcgaggctcccgattcct gtactcttgagaagttcttggggagaattcctatggtgttcaatgtggttatcctttccccccacggatatttcgcccaggaaaatgtcttgggttatcccgacactggtggccaggtgcattactttaatctttatccgtgagtctatgtttgttcgaatcctctctagaaatgtcactgtacct atgtaggatactccaaatataatgcattttggggggatctgttatgggtgcgatggcatttttggaggtcggagcaacaaacaattgctatgtattcttctaaagcttgctttcataaatgctaaggtccttcacccttaatgtgcaggttgtctatatattagatcaagttccagccttggag cgtgaaatgcttaagcgcctaaaggagcaaggacttgatatcacaccgcgtattcttattgttagtatttcctgtacttgtaattactgcggattacacaaaatttcctttttatcttcttaacaacttatcttgatggtattcccaggttactcgtctgctacctgatgcagttggaacgactt gtggtcagcggcttgagaaggtgtatggagccgagcactcacatattctgagggtcccctttaggactgagaagggcattgttcgtaaatggatctctcgctttgaagtgtggccatatatggagactttcactgaggtgacactaaaacttccttatatttgtctatcttctaattggtatta ggaataatttgttaattgttaactctttgtcttttcgtacatcaggatgtcgcaaaagaacttgctgcagaattgcaggccaagccagatttgataataggcaactatagcgagggaaatcttgtggcttcattgctcgctcataagttaggcgtaacacaggtctgtgttgtttttcactctct taaagatctgattgcatttccattagtctggaactagaagtactaaaaagttcttttcttcactgtgttatttgccgtcggcagtgcaccatagctcatgcattggagaaaacaaagttcctgattctgacatctactggaaaaaattcgatgaaaaataccatttctcgtcccagtttaccg ctgatcttattgcaatgaatcacaccgattttatcatcaccagcactttccaggagatagcaggaaggtataacatcaatttgctacttcgactgcaacagcattgtgttcccatttctttcccttatgcttaacctaataccgtcatgaattttccagcaaggacactgtcggacagtacgaga gtcatcaggcattcacaatgcccggattgtacagagttgttcacggcattgatgtgttcgaccccaaattcaacattgtctcacctggagctgacataaacctctatttcccatattccgagaaggaaaagagactgacagcacttcaccctgaaatcgaggagctgctgtacagtgacattga gaacgaggaacatctgtaagtttctaccttactcgtacagtcagtggcggagccagaattttcactaaaataaggtcaaaatataaagacataaatccacaaagaagccaagggtgtcaatatatagtataaatacattaaaaaaattacctatctacacagtgtaattttccgacaaaggggtg tcggttgacactccttgaatacatgtggctctgccactgggtacagttacaaagttctgttacctatgtagatgagcttgtgctgaacatgttgtgattttggcaggtgtgtgctaaaggacaggaataagccaatcttattcacaatggcgagattggatcgtgtgaagaatttaaccggact tgttgagtggtatgccaagaacgcacggctaagggagttggttaaccttgttgtggttggtggagatcgaaggaaagaatccaaagatttggaagagcaaacagaaatgaaaaagatgtatgagctaataaagactcacaatttaaatggccaattcagatggatttcttcacagatgaaccgag tgaggaacggtgaactctaccgatacattgctgacactagaggagctttcgtgcagcctgcattctacgaggctttcggtttgactgttgttgaggccatgacctgtggtttgcctacatttgcaactaatcatggcggtccagctgagatcatcgttaacggaaaatctggcttccacatcga tccatatcacggtgagcaagctgctgatctgctagctgatttctttgagaaatgtaagacagaaccttctcattgggaaaccatttcaacgggtggcctgaagcgcatccaagagaagtaagcaactctttcttgactctagtcattgaaattaactttcttgactctagtcattgaaattaact cgggatttgaggcgtagttgattgatattttatcgcgtctctactgatatatacaggtacacgtggcaaatctactcggagaggctattgacattggctgctgtttacgggttctggaaacatgtttctaagcttgatcgtctagaaatccgtcgatatcttgaaatgttttatgctctca aataccgcaagatggtgagttcctcttcttccttgcccttctcctagtgtttaagatacaatataattgattgcattatcttagagaatcattaatgttaaattttcttaattcttgaatctgttaatgaagtttttctcttggtttttgtttaggctgaagctgttccattggctgctgagtga

SEQ ID NO: 17 : NtSUS2-T polypeptide sequence

MLFMGLKQLSEDFSPAESTAMAERVLTRVHSLRERLDATLAAHRNEILLFLSRIESHGKGILKPHQLLAEFDAIRQDDKKKLNDHAFEELLKSTQEAIVLPPW VALAIRLRPGVWEYVRVNVNALVVEELTVPEYLHFKEELVDGTSNGNFVLELDFEPFTASFPKPTLTKSIGNGVEFLNRHLSAKMFHDKESMTPLLEFLRVHN YKGKTMMLNDRIQNLTTLQNVLRKAEEYLIMLPPETPFSEFEHKFQEIGLEKGWGDTAERVLEMICMLLDLLEAPDSCTLEKFLGRIPMVFNVVILSPHGYFA QENVLGYPDTGGQVVYILDQVPALEREMLKRLKEQGLDITPRILIVTRLLPDAVGTTCGQRLEKVYGAEHSHILRVPFRTEKGIVRKWISRFEVWPYMETFTE DVAKELAAELQAKPDLIIGNYSEGNLVASLLAHKLGVTQCTIAHALEKTKYPDSDIYWKKFDEKYHFSSQFTADLIAMNHTDFIITSTFQEIAGSKDTVGQYE SHQAFTMPGLYRVVHGIDVFDPKFNIVSPGADINLYFPYSEKEKRLTALHPEIEELLYSDIENEEHLCVLKDRNKPILFTMARLDRVKNLTGLVEWYAKNARL RELVNLVVVGGDRRKESKDLEEQTEMKKMYELIKTHNLNGQFRWISSQMNRVRNGELYRYIADTRGAFVQPAFYEAFGLTVVEAMTCGLPTFATNHGGPAEII VNGKSGFHIDPYHGEQAADLLADFFEKCKTEPSHWETISTGGLKRIQEKYTWQIYSERLLTLAAVYGFWKHVSKLDRLEIRRYLEMFYALKYRKMAEAVPLAAE

SEQ ID NO:18 : polynucleotide sequence of NtSUS3-S

atggcgaatccaaagttcacaagagtacctagcatgagggagagagttgaggatactctctctgctcaccgtaaccagcttgttgctctcctctccaggtatattaataaactctatatacttgttattt tctttatttttttgtctttactgataaatttaactgttttcttctttaaatcttgctttcgatgcatgatttctgttgtgttaaattgcgtaaccattttatctaaaagtttatgctgataaacactttt aaattttaatatgtaaattatattatgtctcaacatcaacatgtggatggccaaaaatataaagcttaattttcgttattttgaatgatttttctctgcgagtgttacggtttgcgtacacattacctaa acctcctccctagtccccacttgtgggaatttaatttttttttcttttgtttttttttgttgttgttgttgtctgagttcaattcctaccatgttagcttggcaaaaataagttggtaaagcttgacccc aactagttttagttgatcgatttatttggtgatttatagttcaataataataattactattagagaaagttccagcagcttttctgtttgtttttccagttttagtgattgatatatgtgtatatatatt ctttgtttcttttaagatacgtggcgcaggggaaggggatattgcaacctcaccacttgatcgatgagttcaacaacgctgtatgtgatgacactgcttgtgagaagctcaaagatggtccctttagtga agtcttgaaagctactcaggtatattcactaatccatgggaatcaagatgatactgtatatctttattatggtgtctttcagaaatttgacgatgatgaaatgcaacttttctctgtttgtcaccttatc cagactgtttttttattttttatttttcattttttaacttgaaatgctcttaatttcctttgtttatcgataagaccggatttacaatgtatgaacggagcatcttaagaaccttctggaatgaagatat aagatataaaacatggtgtccgttttctcctttgtggaatcagtgtacatatagactgttattttggtcccactttctggatcttctgatcacaccttctcatgcagaggcgagcttgatggtttcaacc tttaaattcttactattgaatccatttcactttcgaaattatgagttcgaaatctaatatttgttgaaatttttgcaaatgttcacatataagtttaagctttgtgtcaagaatactgggctcaatggat tccaatagaccaggctgtatccgcctctgtctccactctccctgcatccacttctttcgtgtgactaataatgcttaatgagctagaactcgttttaatgtttgaataagttgcttatatcagagcagct tttgatgtttcaatctttaacgggttatgcagtaccagcattctgcggctgaaaaacaggaatctgagatttacttgtctctggctgaatttcttgttcattttgctaacaagtactttggagttaatgc ttgctctctgttgtcaaaataggaagccattgtgctgccaccatttgttgccatagcagttcgtccaaggccaggtgtttgggagtatgttcgtgttaatgtatatgatttgagcgttgaacaattgact gttcctgaatatcttcatttcaaggaagaacttgtggatggagagtaagctctttcttatttcaatacgaaacataaaaatttacagaagttgaataattaacaaatttgttgatttttaatgtatgcca ggggtaataatcactttgtgcttgagctggattttgagccatttaatgcatcagttcctcgtccatctcgatcgtcatccattggcaatggagtccaattcctcaatcgtcatctttcctcaattatgtt tcgcagcaaagactctctggaccccttacttgatttccttagaggacactgtcataaagggaatgtaagtaccaaaagcagttttccctttgtaaatgtctgcttgtccctgattatctactaaatcttt caacacgcgcaaccattataagaaatgtacaatacttctagttagaatttcatcatcgacaaactatctgctttactttttatttttcccatttgatggatgatagtttagtttatataacagatgatat tttggttgaagggtaccatgaactttttcacaaccacttaatggatacatagttgtaatagttgacattttggaataatattgtctcacttggaaatgtttaagaagtattactacttctatttgtaaga tggattgtttatctatgcaggtcttgatgttgaatgatcgtatacagcgaatctccaggctggagtctgctctttctaaagcagaggattatctctccaagctatcaccagatacatcctataatgagtt cgaatacgcgtgagcttgtacacatttgttttgttttctttcaagcatatgtaatttctcaagaaaagggaaatctataggagttgaaacattctttatggaaccatgtgcatgcagattgcaagaaatg ggctttgagagaggttggggtgatactgccagacgtgttttggagacgatgcatcttctttctgacattcttcaggctccggatccatcaaccttggagacatttcttggtagactacctatggtgttca atgtcgtcatattatcccctcatggatattttggccaagcaaatgtcttgggtttgcccgacactggtggccaggtaataacaaggagaatgaggtcttgtattatgtactccctccgttccaatctata tgaacctatttgactgggtatggaaagaaatgaagacttgtaaaacttgtggttctttagaaattccaaacattacatttggttttttccctcttcctggaaattatactactgaatcatctctagatgt tccagtttaacttgagacgtaagggtaaataacggaccattactctgtcctttcttgcagtaggcttggtacaatgaatatagttcgcatagttgccggaagctagagctgtgttagaaaactcaggaac attaatttggcgatgctaatcactgctaatgttactgaagcatccatggttttccttgatgttattctccttttggttgcttcacaggttgtctatatactggatcaagtgcgtgccttggaggccgaaa tgcttcttagaataaagcaacaaggacttaacttcaagcctagaatccttgtcgtgagtacatatatattatgcaagctcttatttggtttgtgggattgcagttgacatcaatttgcttactctgatta ctaaaggtcacacggctgatacctgatgctaaaggaaccatgtgcaaccagaggttggagaggattagtggaactgaatactcgcatattttacgtgtcccttttaggacagagaagggaatccttcata aatggatatctaggtttgatgtatggccttacctggagaagttcactgaggtaacctctttgtcccttggaaattgccttttgttgctgatgtttctgctagtgtgcttaaatgacggatgttaactagt cacttgctagcgtttgcaatagcaacgggaaaagaaaggatttttgctagtttgaagtctgcctccaagaaaaattatattaaaagtttatggctagtggaaacatcagtcattcatgtaccttatttct atgcccaagttgtttaagttgaaagtaatttggccaactatgcaaattggggagaacgtgtagccaactattgtgtttgccgacatgttgatatactttttggtcctgatttatatttgttggtttgtcat actggatgaagcaattctcatgtttttctgcttatatatattggaagaagagatacttgtcgtttcatcatttttctcgacctctctattaccaacactttgccaatttaatgtttggaaatgtcttctt gaccaggatgtggcaagtgaaatgaccgctgagctccagggaaagccagatctgattattggcaactacagtgatggaaatttagttgcctcccttttggcatataaaatgggtgtcacacaggtaggaaa tacatgattctttatcttgctagcactaagtcttgaggttatgtatctgcaatagaaattttacgctttgccttcatttctttttaattatttttccagtgtaccattgctcatgccttggaaaaaacaa agtatcctgattctgacatctactggaaaaagtttgaggagaaatatcatttttcatgtcagtttactgctgatctactggcaatgaataattcagatttcattatcaccagtacttatcaagagattgc aggaacgtaagtcattttaatctggtcgtttaaatctgatatttcttccctagtagtctattcaatccgaatttcagttcagtatatgatgtcatcggttgaggaactgtgattggtaaccttatcaaat ccgtagctgctctataattttatttcgtaattggagaaacaattttttattattgagcttgtagtctgagctagaatttggttctttatctatcaagtagcataatactacaactattttttatgtgtgg caatttgcaatttcaattttctatttctataagttgcagcttttcttcctgttctgatcatatttacatggctgaaactcaatagaaaactaggctagttgatcaaaagtagttggatgctttaaaatta gtagacgttttgctaaatgagtgaccaatgttattaaaaaaacgttcatgttttcaacccttttggcatacatttgaccactgcccaagattttggataagtacatgcagtgcttataattataaagcat tttatcccaccttgtttttcattatgaaaattaagtaatttacgagtatttgtataagttacttcataaattagaagtaaatctggattgtgtaaagttattcgccccgtatatactgaaagctacttga acaagcaaaaaaacagacaaacgtaacattctccatggattaatgagacttgtatatatatatatatatatatgtaaagagagagagagagatttggcttgtaaccacatgtatattatgccatatgg atgtgacattgatgtgactagacctaaatgttttgtttcaatgtccacgggagttttacgtagagttaagaggagaagagagtgaggaatactaatgtttgatggtaccccttggcttcttgacctggat actcagtgttcttattcatgcctatactttggtccttgatttcattctcccttttctagcttgagctgcatcaaagaaattccactgtaaaaaaaataatgctcaccatattggtgcaacatggcaaaca tgtatcctatttgatgatcaatcaactttatttttctcctgttaattgacctcagtgtgtaactctctatgtatgatagcattgtaacttgtgtcatgattcataaatagggtactagaattggatggtt gacatagtaaatggtcaattgatgatccacaaaatatgcacctactgattaaaatgtgatagggcaggtttatttttgtttgtggttaacacagtacttaaccctatatttaatacaatttggcttatct acaatcttttcttcagtgtttatgcgaattccttattgcacaacaatattgtctttctgagttctattctgttgttgcttacacttttattattccagtaacatagatgtgaagacattagattggttgc ttgcaaattgatagccacttgtttcaggaagaatactgttggtcagtacgagagccatactgcattcaccctcccgggactatatcgcgtcgttcatggcattgatgttttcgatcccaaattcaatata gtgtctcctggagctgacatgacaatttatttcccatattctgacaaggaaaaaagactaacgtctttgcatggctcgattgaaaagttgttatttgatcctgcgcagaatgaagagcatatgtaagtgg catccgtttgtacttaatttttttggaatagatgacatattatttgcatgaatatgaaaaggagggtctgatatgattttctatagataaactaccaatgatattatttaaaaactcctggatactgtatt aggagaagaagagaaccaggggtagatggcattagaatcccttaaatcttgaagagtcgtcactaacgctcccaacacttctgcctcagaccctcaactaaatactattattgttgatttctttggagaa gctataagaatctctctccttatggtgaaaattttacttggctttatacttaacttccaaggctccctcttataaaatgcaaaaactgtctgtattcactctcttggttaacaattgatccaatcaaa tgcatatggaacatctttctttacgtttcttctaaagttcgtttgaggataaggagtagaatctgagaagatagactagtaggtaaccttagggacggatgtggaaattaacatatgggctcagcttttc tgccgagtgcagaccatgtatatgcgttaaaaaattcactaaacaagtaaatgtttgattttgaacccagtaaatcaaatgagttgtggtagaatctcgaactcgaaccgataaagttcaaatccaggat ccgcttttaggtaaactctaccttgggaagtgttatatatatgtccctgattatttctttttccgtttcctttctattttaatttttaaagttatttttagatggttttattttttgataagtggtaagt tgttaatattccaaattaaatgccattgtcataactatatacattttataaagaatgattgatcctagtttctcattcctaagatccaaataaggcaataaacaatgtcttagtaattggacctgcttctg gtgatcaacgcttgatcgcgtagttagttatagatgactgtaaaaactttaaccattttaatggttttgtcaaagaacaaatatcggacatattatagagaatggactattgtactttgcttctgattgg tcattttattgtgatccgtaaattggctgtgactgatgtcatatctttgcttacagaggtaatctgaatgataaatcaaaacccataattttttcaatggcaaggctagaccatgttaagaacattacgg gactagttgagtgctatgctaaaaatgccacattgagggaattggcgaaccttgttgtagtagctggatacaacgatgtaaagaaatccagtgatagagaagaaataacagaaattgagaagatgcatgc tcttattaaggagcataaattggatgggcaattcagatgggtatcagcccaaacaaaccgggcacgtaatggtgagctctatcgctatatagctgaccagagaggtatatttgttcaggtatgctatttg tattgtattagtccaatttcattttttgcaccaaaagaaaggttgttattgtgacgtatatgtttgttttagcctgcattttatgaagcatttggactaacggtggttgaagctatgacttgtggtcttc caacatttgcaacttgccatggtggtcctaatgagatcattgaacccggtgtatctgggttccatattgatccttatcatcccgataaagctgctgaactcatgtcagaattctttcaacgctgcaaaca agatcctactcactgggaaaaaatatctgcatctggtctccgaaggattcttgagaggtctgtagttgtgtacatgtatagaagattaaagaatgctaccttgatatttatttgaatcaaaaataacagg aacatctcttttttgaacatcactcaagttcttatattaaataatttttaggtatacgtggaagatttactccgagaggctgatgactttatctggcgtatatggtttctggaagcttgtttcaaaactt gagaggcgtgaaactagacgataccttgagatgttctacattctcaaattccgcgagttggtgagtgccttttagctccttttcagttccaataaactatatatgtggtttaagtaagtattaagcataa acatgtccgtgcttggggctgtcgaaaatgctatggacatatcctgagctaaggatttttcaagaaaattgatgttagctttactctatttacaggcaaaatctgtacctctagcaattgatgacaagtga

SEQ ID NO: 19 : NtSUS3-S polypeptide sequence

MANPKFTRVPSMRERVEDTLSAHRNQLVALLSRYVAQGKGILQPHHLIDEFNNAVCDDTACEKLKDGPFSEVLKATQEAIVLPPFVAIAVRPRPGVWEYVR VNVYDLSVEQLTVPEYLHFKEELVDGEGNNHFVLELDFEPFNASVPRPSRSSSIGNGVQFLNRHLSSIMFRSKDSLDPLLDFLRGHCHKGNVLMLNDRIQR ISRLESALSKAEDYLSKLSPDTSYNEFEYALQEMGFERGWGDTARRVLETMHLLSDILQAPDPSTLETFLGRLPMVFNVVILSPHGYFGQANVLGLPDTGG QVVYILDQVRALEAEMLLRIKQQGLNFKPRILVVTRLIPDAKGTMCNQRLERISGTEYSHILRVPFRTEKGILHKWISRFDVWPYLEKFTEDVASEMTAELQ GKPDLIIGNYSDGNLVASLAYKMGVTQCTIAHALEKTKYPDSDIYWKKFEEKYHFSCQFTADLLAMNNSDFIITSTYQEIAGTKNTVGQYESHTAFTLPG LYRVVHGIDVFDPKFNIVSPGADMTIYFPYSDKEKRLTSLHGSIEKLLFDPAQNEEHIGNLNDKSKPIIFSMARLDHVKNITGLVECYAKNATLRELANLV VVAGYNDVKKSSDREEITEIEKMHALIKEHKLDGQFRWVSAQTNRARNGELYRYIADQRGIFVQPAFYEAFGLTVVEAMTCGLPTFATCHGGPNEIIEPGV SGFHIDPYHPDKAAELMSEFFQRCKQDPTHWEKISASGLRRILERYTWKIYSERLMTLSGVYGFWKLVSKLERRETRRYLEMFYILKFRELAKSVPLAIDDK

SEQ ID NO:20 : polynucleotide sequence of NtSUS3-T

atgtttacatggctgaaactcaatataaaaaacaagggtaggtgatcaaaaatcgttggatgcttaaaatcagtagacgttttgctaaatgagcgaccaatgttattgaaaa cgttcatgttttcaacccttttggcatacatttgagcattgcccaagattttggataagtagatgcagtgcttataattttaaagcattgtatcctgccttgtttttcattgt caaaattaattaacttacaagtatttctataagttgcttcataaattagaagtaaatctggattgtgtaatgttattcgcctcgtaaatactgaaagctgcttgaacaagtg aaaaaacacagacaaacgtaacattctccatggattgatgagacttgtaaaatacatatatagaaatttggcttgtaaccacatgtatattatgccatatggatgtgacattg atgtgactagacctaaatgttttgtttccatgtccactggagttttacgtatagttaagaggagaaaagactgaggaatactaatgtatgatggtacccctttgcttcttga cctggatacccagtgttcctattcatgcctatactttggtccttgatttcactctcccttttctaacttgagctgcatcaaagaaatttccactgtaaaaaaataaataatgc tcaccatatctctgcaacattgcaaacatgtatcccatatgattgatattggtgcgacatggcaaacatgtatcctatttgatgatcaatcaaatttatttttcccctgtcaa aatgacctcagtgtgtaattccctatgtatttgatagcattgtaactcgtgtcatgattcatgaatagggtactagaattgcatggttgacaaatattaactggtcgattgat gatccacaaaacatgcacttactgactaaaatgtgatgggacagatttatttttgtttgtgattaacacagtacttaaccctatacttaatacaatttggcctagctacaat cttttcttcagtgcaaattccttgttacacgaccaatattgtctttctgagttctattctctgttgttacttacacttttattattcgaataagacattagattgcttgcatgca aattgatagccacttgtttcaggaagaatactgttggtcagtacgagagccatactgcattcaccctcccaggactatatcgcgtcgttcatggcattgatgttttcgatcc caaattcaatatagtgtctcctggagctgacatgacaatttacttcccatattctgacaaggaaaaaagactaacgtctttgcatggctcgattgagaagttgttatttgatc ctgcgcagaatgaagagcatatgtaagtgacatccattgtacttattttaatttggaatagatgacatacttatttgcatgaatataaactgacaacccagagatttccta cattagaaaaggagggtctgatatgattttctacaaataaattcccagtgatattgttcaaaaagtcctggatactttattatgagagaaccagggatagatggcactagaat cccttaatcttgagaagtcgccacttatcgctcccaacactttctgagaccctcaagtaactactattattgtttgatatcttggagaagctataagaatctttttctcctta ttgtaattttttttacgtgactttaaacttaacttccaagctccttctgataaaatgcaaaaactgtctgtattcactgtcttggtttattaacaattgatccaatcaaatgc atatggaacatctttctttttgtttcttcaaaagttcgtttgaggataaggagtagaatctgagaagatagactagtaggtaaccttaggggcggatgtagaaatcaacgta tgggttcagctttgttgcagaccctgtatatgcattaaaaaaatcactaaataagtaaataattgattttgaacccagtaaatcaaaatgagttgtagtagaatcctgaactc gaaccgataaagttggatccactaccgggtaaactctaccttgagaagtgtttatatatgtccctaattatttcttttctgtttcctttctattttaattttttaagttcct ttttagatggttttattttttgacaagtggtaagttgttagtattccaaattaaatgccattgccataactatatacatttataaagattgattgaccctagtttctcattcc taagatccaaataaggcaataaacaatatgtcttagtacttgaacctgcttctggtggtcaacacttgatcgcgtagttagttatagatgactgtaaaaaccttaatcattt taatggttttgtcaaagaacaaatatcggacatattatagcgaatggactattgtacttttcttctgattggtcattttattgtgatccgtaagttggctgagactgatgtca tatctttgcttacagaggtaatctgaatgataaatcaaaacccataattttttcaatggcaaggctagaccatgttaagaacattacgggactagttgagtgctatgctaaaa atgccacattgagggaattggctaaccttgttgttgtagctggatacaacgatgtaaagaaatccagtgatagagaagaaatagcagaaattgagaagatgcatgctcttatt aaggagcataaattggatgggcaattcagatggatagcagcccaaacaaaccgggcacgtaatggtgagctctatcgctatatagctgacaagagaggtatatttgttcagg tacgctgtttgtattgtatttgtccacattcctttttttgcaccgaaagaaaggttgttattgtgacaaatatgtttgttttagcctgcattttatgaagcatttggactcac ggtggttgaagctatgacttgtggtcttccaacatttgcaacttgccatggtggtccgaacgagatcattgaacacggtgtatctgggttccatattgatccttatcatccc gataaagctgctgaactcatggcagaattctttcaacgctgcaaacaagatcctactcactgggaaaaaatatctgcatctggtctccgaaggattcttgagaggtttgtagt tgtgtacatatatagaagattaaagattgttcccttgatattatttgaatgaaaaataacagtaacatctctttttgaacatcgctcaagttcttgtgttaaataattgtta ggtatacgtggaaaatttactccgagaggctgatgactttgtctggtgtatatggtttctggaagcttgtttcaaaacttgagaggcgcgaaactagacgataccttgagatg ttctacattctcaaattccgcgagttggtgagtgcctttttgctcattttcagttacaatcaactatatatgtggtttaaatacgtattaagcataaacatgtccgtgattgc ggctgtcgaaaatgctatggacatatcctgagctaaggagttttcaagagaattgatttggcttactctgtttacaggcaaaatctgttcctctggcaattgatgacaagtga

SEQ ID NO:21 : NtSUS3-T polypeptide sequence

MFTWLKLNIKNKGRKNTVGQYESHTAFTLPGLYRVVHGIDVFDPKFNIVSPGADMTIYFPYSDKEKRLTSLHGSIEKLLFDPAQNEEHIGNLNDKSKPIIFSMARLDHVKNITGLVECYAKNATLRELANLVVVAGYNDVKKSSDREEIAEIEKMHALIKEHKLDGQ FRWIAAQTNRARNGELYRYIADKRGIFVQPAFYEAFGLTVVEAMTCGLPTFATCHGGPNEIIEHGVSGFHIDPYHPDKAAELMAEFFQRCKQDPTHWEKISASGLRRILERYTWKIYSERLMTLSGVYGFWKLVSKLERRETRRYLEMFYILKFRELAKSVPLAIDDK

SEQ ID NO:22 : polynucleotide sequence of NtSUS4-S

atggcggaacgtgtgctgactcgtgttcatagccttcgtgaacgtcttgatgctactttggctgctcatcgcaatgagattttgctgtttctttcaaggtatagtcttagcagattgttctttga tttagttgttattgccagttctaatgtatgggcttatatataaacaaagtgttgaagtatgcaaccatataaactgacagcttaaaatgcttgagagaacacacttttatttatttaattatgcct tcagcacaagaagtggaacttgacgcaatggaaccataggtcacgggttcaagtcttggaacagcctgcaatctaaggctgcgtgtagtagaccctagtggtccggcccttccacatatctcgctt agtgtaccgggcccattgagtacgggttcggccgaacccagtcgctttggtccaatccatatatttgtcttaaaaatatattgaatatatacaaattgttaatttagtttaaatatgtgtatcatg ggttattcatgctggttttggctgttgcaggattgaaagccatggaaaagggatactgaaacctcaccagttgctggctgaatttgattcaattcacaaagaagacaaaaacaaactgaatgatca tgcttttgaagaagtcctgaaatccactcaggtatttgtggttttagtgttaggtgatggatagcatttattgttttactaagatcacatatgtgtcagtttgtggctagtatttaaaatctggtg tattttgtcatactaggaagcaattgttttgtccccttgggttgcgcttgccattcgtctgaggcctggtgtgtgggaatacgttcgtgtgaatgtcaacgctcttgttgttgaggagcttaccgt gcctgagtatttgcaattcaaggaagaacttgttaatggaacgtaagttttaggttcgaatttgttgatttgttagataacatgttctgaactttttgattaaagttgtgtttttgactgatgcag ctcgcacgataactttgttcttgagttggattttgagcccttcactgcatcatttccaaaaccaaccctcaccaaatcaattggaaatggagttgaattccttaaccgacacctctctgccaaaat gttccatgacaaggaaagcatgacccctcttctcgagtttcttcgagttcaccactacaagggcaaggtaaacttgtttttcctgtttgtctatgaatttagtttagttgttttgctccgcgaaaa tttcagtggaaactgatttatgcaaccactgagtgattaatatgttcaaacttaccgacttctggttttctgtgtagacaatgatgctgaatgacagaattcaggacttaaatactctccaaaatg tcctaaggaaagctgaggaatacctcactaccctttcccctgaaacttcatactcggcatttgagcacaagttccaagaaattggcttggagaggggttggggtgacactgcggagcgtgttctag agatgatctgcatgctcctggatctcctcgaggctcctgactcgtgcacgcttgagaagttccttggtagaattccaatggtttttaatgtggtcatactttcaccccatggttatttcgcccagg aaaatgtcttgggttaccccgacactggtggccaggtgcactgcttatctgtgttcggtcttattatctctttaaaccctactgccacaagtgctgagatgaacctcctttaatttgcaggttgtc tatattttggatcaagttcctgctttggagcgtgagatgctcaagcgcataaaggagcaaggacttgacatcaaaccgcgtattcttattgttcgtattcccagtaattgtgtttaaacttatgat tatgcaggattttatctgttctaatacagcactcttgcttaaattctcaggttactcggctgctgcctgatgcggttggtaccacttgtggtcagaggcttgagaaagtgtttggaacagagcact cacacattcttagggtcccctttaggaccgagaagggcattgttcgcaaatggatctctcgctttgaagtctggccatacatggagacattcactgaggtgaagcaagctttctctattcatttt tcaatcttccaattggttttggcagcaattttctgcttgctttgacttccgctaaaacttcggattttattgcattaggatgtggcgaaagaaattgctgcagaattgcaggctaagccagatctt atcattggcaattatagtgagggcaaccttgctgcctccttgttggctcacaaattaggtgtaacacaggtcggcaatgtttgtgacatgtaatttcatctttgcatttcctttcgtttgcaacta aaagattaagagttctctctctcttttttttttccgtctactttgccttatgcagtgcacgatagctcatgctttggagaaaacaaaatatcctgattctgatatctacttgaagaaatttgatg aaaaataccatttctcagcccagtttactgccgatcttattgcaatgaatcacaccgatttcatcatcaccagcactttccaggagatagcgggaaggtatttttacatcagtttcccactctgat taaattacaatgtatttccctatatgattaaatactgtgtttgatcctaaatcatttctaaattttccagcaaggacactgttggacagtacgagagccacatggcgttcacaatgcctggactgt atagagttgttcacggcattgatgtgtttgaccccaaatttaacattgtgtcaccaggagctgatatgaatctctatttcccatactacgagaaggaaaagagattgacagcatatcaccctgaaa ttgaggagctgctgtttagtgatgttgagaatgacgaacacatgtatgttactaaactagcaatcctgctgcaaaattatggctaattatgtaaacaagtttgtactgaatagatttgttattcga tcaggtgtgtgctgaagaacaggaataagcctatcatattcactatggctagattggatcgagtgaagaacttaactggacttgtcgagctgtacgccaagaacccacggctaagggagttggtta accttgtcgtggttggaggagaccgaaggaaagaatccaaagacttggaagaacaggcagagatgaagaagatgtacgaacttataaagactcacaatttgaacggccaattccgatggatttctt cccagatgaaccgcgtgaggaatggcgaactctacaggtacattgccgatactaggggagctttcgtgcagcctgcattttacgaggcttttggtttgactgttgttgaggccatgacctgtggtt tgcctacatttgcaactaatcacggtggtccagctgagatcatcgttcacgggaaatctggtttccacattgatccataccacggggatcaggcagctgaacttctcgctgatttctttgagaaat gtaagaaagaaccttcgcactgggaagccatttccgagggcggccttaagcgtatacaggagaagtaagcaaactgctactcttttcatttttgcaaaacctactatgatcattattaagctcatt tttgcaaaacctacttgctgttgttattgtttgttgcttccttttcactgttctttgagctgaaggtctatcagaaacagtctctctaccttcacaaggtaggggtaagatctgcgtgcacgttac cctcctcaaactctacttaattgtgagattacactaggtttgttgttgttgattctttgctaattaattaaaaggtacacatggcaaatatactcggatcggttgttgacactggctgctgtatat ggattctggaagcatgtttccaagcttgatcgtcttgaaattcgccgttatcttgaaatgttctatgctctcaaattccgcaagctggtgagtttcattgctttctgcactcctgcaattgtatag

SEQ ID NO:23 : NtSUS4-S polypeptide sequence

MAERVLTRVHSLRERLDATLAAHRNEILLFLSRIESHGKGILKPHQLLAEFDSIHKEDKNKLNDHAFEEVLKSTQEAIVLSPWVALAIRLRPGVWEYVRVN VNALVVEELTVPEYLQFKEELVNGTSHDNFVLELDFEPFTASFPKPTLTKSIGNGVEFLNRHLSAKMFHDKESMTPLLEFLRVHHYKGKTMMLNDRIQDLN TLQNVLRKAEEYLTTLSPETSYSAFEHKFQEIGLERGWGDTAERVLEMICMLLDLLEAPDSCTLEKFLGRIPMVFNVVILSPHGYFAQENVLGYPDTGGQV VYILDQVPALEREMLKRIKEQGLDIKPRILIVTRLLPDAVGTTCGQRLEKVFGTEHSHILRVPFRTEKGIVRKWISRFEVWPYMETFTEDVAKEIAAELQA KPDLIIGNYSEGNLAASLLAHKLGVTQCTIAHALEKTKYPDSDIYLKKFDEKYHFSAQFTADLIAMNHTDFIITSTFQEIAGSKDTVGQYESHMAFTMPGL YRVVHGIDVFDPKFNIVSPGADMNLYFPYYEKEKRLTAYHPEIEELLFSDVENDEHMCVLKNRNKPIIFTMARLDRVKNLTGLVELYAKNPRLRELVNLVV VGGDRRKESKDLEEQAEMKKMYELIKTHNLNGQFRWISSQMNRVRNGELYRYIADTRGAFVQPAFYEAFGLTVVEAMTCGLPTFATNHGGPAEIIVHGKSG FHIDPYHGDQAAELLADFFEKCKKEPSHWEAISEGGLKRIQEKYTWQIYSDRLLTLAAVYGFWKHVSKLDRLEIRRYLEMFYALKFRKLVSFIAFCTPAIV

SEQ ID NO:24 : polynucleotide sequence of NtSUS4-T

atggccgaacgtgtgctaactcgtgttcacagccttcgcgaacgtcttgatgctactttggctgctcatcgcaatgagattttgctgtttctttcaaggtatagtcttagcagattgttctttgat ttagttggtgttatttgccagttctaatgtatggactaatatatgaacaaagtgcgaccatttcaactgacaacttaaaatgtttgagagaatacacgtttatttacttaattatggcttgagcata ggaagtgtatcttggcgtaactcgtaaagttgacctcatgtgacaaggaggtcacggtttcgagccgtggaaacagcctcttgcagaaatgcaggtaaggctgcgtgcaatagatcgcccttccac ggacccgcgcatagcgggaacttagtgcaccggttgggctgtcctttttttatgtcttcagcacaaaaatttagtttaaacatgtgtatcatggattattcatgctggttttgccggttgcaggattg aaagccacggaaaagggatattgaaacctcaccagttgctggctgagtttgaatcaattcacaaagaagacaaaaacaaactgaatgatcatgcttttgaagaagtcctgaaatctactcaggtaa tttgtggttttagtgttaggtgatggatagcatttattgtcttactaagatcatatatgtgtcagtttgtggctagtatttgaaaagtctggtgtggtttgtcatactaggaagcaattgtcttgtc cccttgggttgcgcttgccattcgtctgcggcctggtgtgtgggaatatgttcgtgtgaatgtcaatgcacttattgtcgaggagctgactgtgcctgaatatttgcaattcaaggaagaacttgtt aatggaacgtaagttttaggttcgaaatgatgatttgttaaataatatgttctgaactttttgattaatgttgtgttttcccctgatgcagctcgaacgataactttgttcttgagctggattttga gcccttcactgcatcatttcccaaaccaaccctcaccaaatcaattggaaatggagttgaattcctcaaccgacacctctctgccaaaatgttccatgacaaggaaagcatgacccctcttctcga gtttcttcgagttcatcactacaagggcaaggtaaacttgtttttcctgtttgtctatgaatttagtttctgaaagttgctttgcttcgtgaattttttagtggcaactgatttatgattttctgtg cagacaatgatgctgaatgacagagttcaggacttaaacactctccaaaatgtcctaaggaaggctgaggaatatctcactaccctttcccctgaaacttcatactcggtatttgagcacaagttcc aagaaattggcctagagaggggctggggtgacaatgctgagcgtgttctagagatgatctgcatgctcctggatctcctcgaggctccagactcatgcactcttgagaagttccttggtagaattcc tatggtttttaatgtggtcattctttcacctcacggatatttcgcccaggaaaatgtcttgggttaccccgatactggtggccaggtgcactgcttatttgtaacaccttacgcttttccctctga aacttatttgcggcaagttctaaggtcctccttccttaatttgcaggttgtctatattttggatcaagttccggccttggagcgtgagatgctcaagcgcataaaggagcaaggacttgatatcaaa ccgcgtattcttattgttcgtatctccaataattgcgtttaaacttatgattgtgcaggatttgatctgttcaaatctaatgactgattttctttttttttttttttccctcaggttactcggctg ctgcctgatgcggttggtaccacttgtggtcagcggcttgagaaagtgtttggaacagagcattcacatattcttagggtccccttaggaccgagaagggcatcgttcgcaaatggatctctcgct ttgaagtctggccttacatggagacattcactgaggtgaagcaagctttctctattcatttttcaatcttccaatctgttttggcagcaatttttcacttactaacactttggctttcgctaaaac ttcggattttattacattaggatgtggcaaaagaaattgctgcagaactgcaggcaaagccagatcttataatcggcaactacagcgagggcaaccttgctgcctccttgttggctcacaagttagg tgtaactcaggtctgtaatgtttgtcacctgttatttcaactttgcatttcctttcatttgcaactagaagttaagagttctctcttttatcttttccgtctattttgccttctgcagtgcacc atagctcatgcgttggagaaaacaaaatatcctgattctgatatctacttgaagaaatttgatgaaaaataccatttctcagcccagtttactgccgatcttattgcaatgaatcacaccgatttca taatcaccagcactttccaggagatagcgggaaggtattacatcacaatggatttccgatatgattaaattagttaatttaatcctacttcattgtgtttgatcctaaaacttttctaaatttccc agcaaggacactgttggacagtacgagagccacatggctttcacgatgcctggattgtatagagttgttcacggcattgatgtgttcgatcccaaattcaacattgtgtcaccaggagctgatatga atctctatttcccctacttcgagaaggaaaagcgattgacagcatatcaccctgaaattgaggagctgctgtttagcgatgttgagaatgacgaacacatgtatgttactaaactagcaatcctgct gcaaaattgtggctaattatgtaaaaaagtttttactgaatagatttgtgcttctatcaggtgtgtgctgaaggacaggaataagccaattatattcaccatggctagattggatcgagtgaagaac ttaactggacttgtggagttgtacgccaagaacccacggctaagggagttggttaaccttgtcgtggttggtggagaccgaaggaaggaatccaaagatttggaagaacaggcagagatgaagaag atgtatgaacttataaagacgcacaatttaaacggccaattccgatggatttcttcccagatgaaccgcgtgaggaatggcgaactctacaggtacattgccgatactaggggagcttttgtgcagc ctgcattttacgaggcttttggtttgactgttgttgaggccatgacctgtggtttgcctacgtttgcaactaatcacggtggtccagctgagatcatcgttcacgggaagtctggttttcacattga tccataccacggcgagcaggcagctgaacttctagctgatttctttgagagatgtaagaaagaaccttcacactgggaagccatttccgagggcggccttaagcgtatacaggagaagtaagcaagc tgctactcttttcatttttgcaaaacctaccatgatcattattaagctcatttttgcaaaacctacttgttattctttgttgcttccttttccctgttttttgagccgaggttttatcgaaaacat gctttctaccttcacaaggtaggggtaaggtctgcgtttgttattattgttgttgttgattctctgcgaattaattaaaaggtacacatggcaaatctactcggatcggttgttgacactggctgct gtttatggattctggaagcatgtttccaaacttgatcgtcttgaaattcgtcgttatcttgaaatgttctatgctctaaaattccgcaaactggtgagtttcactgctttctgcactcttccaattg ttagttgagtgcactcatttaaactgtagctaaagctgttgtaaatcttcagttaagcagctgctaatgaagtttttatcttttgtttttggttcaggctgaagctgtcccgttggctgttgagtaa

SEQ ID NO:25 : NtSUS4-T polypeptide sequence

MAERVLTRVHSLRERLDATLAAHRNEILLFLSRIESHGKGILKPHQLLAEFESIHKEDKNKLNDHAFEEVLKSTQEAIVLSPWVALAIRLRPGVWEYVRV NVNALIVEELTVPEYLQFKEELVNGTSNDNFVLELDFEPFTASFPKPTLTKSIGNGVEFLNRHLSAKMFHDKESMTPLLEFLRVHHYKGKTMMLNDRVQDL NTLQNVLRKAEEYLTTLSPETSYSVFEHKFQEIGLERGWGDNAERVLEMICMLLDLLEAPDSCTLEKFLGRIPMVFNVVILSPHGYFAQENVLGYPDTGG QVVYILDQVPALEREMLKRIKEQGLDIKPRILIVTRLLPDAVGTTCGQRLEKVFGTEHSHILRVPFRTEKGIVRKWISRFEVWPYMETFTEDVAKEIAAEL QAKPDLIIGNYSEGNLAASLLAHKLGVTQCTIAHALEKTKYPDSDIYLKKFDEKYHFSAQFTADLIAMNHTDFIITSTFQEIAGSKDTVGQYESHMAFTM PGLYRVVHGIDVFDPKFNIVSPGADMNLYFPYFEKEKRLTAYHPEIEELLFSDVENDEHMCVLKDRNKPIIFTMARLDRVKNLTGLVELYAKNPRLRELVN LVVVGGDRRKESKDLEEQAEMKKMYELIKTHNLNGQFRWISSQMNRVRNGELYRYIADTRGAFVQPAFYEAFGLTVVEAMTCGLPTFATNHGGPAEIIVHG KSGFHIDPYHGEQAAELLADFFERCKKEPSHWEAISEGGLKRIQEKYTWQIYSDRLLTLAAVYGFWKHVSKLDRLEIRRYLEMFYALKFRKLAEAVPLAVE

SEQ ID NO:26 : polynucleotide sequence of NtSUS5-S

atggcctcaacagttgctgatagcatgcctgatgctttgaaacaaagccggtatcatatgaagagatgcttcgctaggtgaacacccttcttttatgttttttcccctctacgtgtt tatgtcaaatttccatgcataatgctaactacttttcttctttttgacttcaaaattggatgtgaaaggttcattgcaatgggaaggaggctaatgaagttgaaacatttaacagaa gaaatagaagaaactattgaagacaaggcagaaagaaccaggattttggagggttcacttggaaaaattatgagttccacacaggtcagcaccatttaaccaacttagttgaacagg aaaaaaagaaaaagcaaaagagttattgcaaggcgtaacgattttctttgaaattttcaggaggcagctgttgttccaccttatgttgcttttgcagtaaggcacaatcctggcttct gggattatgtcaaagttaacgctgaaactctctctgtggaagctatttcagccagggaatatctcaaattcaaagagatgatctttgacgaagactggtaagtggaaaattgtatca ttttaaagagaaacaattttgtaacatacaagaatagttttgatggttgaatgtgcaagcagggcaaaggatgataatgcactggaagtagattttggtgcttttgactactctaat cctcggttagccctttcctctcttctgtcggaaatgggctcaactttatctcaaaagttctgtcttcaaagtttggtggaaagccagaggacgcccagcctttgcttgattacttacta gctcttaatcatcaaggagaggtatgaaaatggactacctttgtttcttaaaggtattatataatgatgcgcgttataaagttcctttttaaattgaaactttgcagaatctaatgat caatgagaatctgaatggtgttgctaagcttcaagcagcattgatagtagctgaagtttttgtatcttcctttcccaaagacacacctttataaagactttgagcataagtaagcttc tcatatgcttccattgtcatatgcagtataccaatgacatgctaccgaaaagttgtttatgtttgtgacttgattatgaaaactctaggctcaaagaatggggctttgataaagggt ggggtcacaatgcaggaagagtaagagagacaatgagactgctttccgagataatccaagcaccagatcccataaatatggagtcctttttcagcaagcttcctactacattcaaca ttgttatcttctccattcatggttactttggccaagcagatgtccttggtctgcccgatactggaggccaggtctacatatacagcaatttatctccttttgcctcatattgcttatt agcgacacttgcatcattgaaatcagacttttacttcacaggttgtttatattctggatcaagtaagggcttttagaggaggaaatgttacaaagaatcaagcagcaagggctaaacg tgaagcccaagattcttgtggtgagttttgcaaaaatatgcttagacaggttttgagattgatcggagaagggattaagatgatcaagatctttgtttcctgctttcatgatgtaaa caggtatctcgtctcataccagatgctcgagggacaacatgcaatcaggagatggaacctattcttaactcatcccattctcacatcctgagaattccattcaggactgagaaagga gttcttcgccaatgggtttctcggtttgatatctatccttacttggagaactatgccaaggcaagtcttctaacaaaattaccacctattcatacactttatttactttcttgaacta atcgtttggtttgtgacgtatatcattaggatgcttctgctaagatacttgagctcatggaaggtaaaccagacctcataattgggaactacactgatggaaatttagtggcatctc tattggccaacaaacttggagttactcaggttccgtagctgatcatatgatcatattttctacattgtttcttgataattaaatggaaatcttattggatgataacattttagggaa ccattgctcatgcattagagaaaactaagtatgaagattctgatgtgaagtggaagcagtttgatcccaagtaccacttttcttgccaatttactgccgatttattggcaatgaatg ctgctgattttatcattaccagcacatatcaagaaatcgctggaaggttagcactgactctctcagtatatttggcaacttaatgaatttactgcagtggccaacactaaaagctatc attcgtccttcagcgaaactaggcctggacaatatgaaagtcacacagcatttaccatgccggggctttatagagctgtttcaggcatcaatgtatttgatccaaagttcaacattg ctgctcctggggctgaacagtctacctatttccctttcactgagaaacagaaacgattcagcacatttcgtcctgctattaacgaattactttacagtaatgaggaaaacaatgagc acatgtaagtctaattgcccattttcctaatctaaccattgcttaaatcgttctgtttttaccggatgtgtggtacttatcagtaacatttttttttggatcagtggatttcttgca gaccggaaaaaaccaattatattttcaatggcgagatttgatacagtgaagaacctgtcaggcttgactgagtggtatgggaagaataagaagttgcggaacttggtaaaccttgtta ttgttgggggattcttcgatccatcaaaatcaaaagaccgggaggaagcagctgaaatcaagaagatgcatgaattgattgagaaataccagctcaagggacaaatgagatggatag cagctcaaactgataaatatcgaaatagtgagctataccgaactattgctgacactaagggagcttttgtccaaccggctttatatgaagcttttggactaaccgttattgaagcaa tggattgtggattgcctacgtttgcaactaatcaaggtggacctgcagaaatcattgttgatggggtttcaggtttccatattgatccttacaatggggacgaatcaagcaagaaaa tagctgatttctttgagaagtgtaaggttgattctaaatattggaacaggatatctgagggaggtctcaagcgcattgaagaatggtaacaaactagttccaagtttaaaaaatggaa aaaatgcttatcatgttatattttcgtggttttaagttctgcttcgatgcagttatacgtggaagatttatgcaaacaaagtgttgaatatgggatcaatctatggattttggagac aattcaatgtggggcaaaagcaggctaagcaaagatactttgagatgttttacaatcctctcttcaggaaattggtaggttgtatatgttgaatacaatttactaagatcctcaaaa tgaccaagaaatatacattgactatgctacttttgtaatttcacaggccaaaagcgtgccgatcccacatgaagagccattgccacttgcaacatcagactctactcaatcccaaga attaaaactaccactaccagttccagcagcagtagctaaagttctgccattaacaaggcatgcttttaacttaattacttctctacctagagtaactggtaaagtggatgtcaagtga

SEQ ID NO:27 : NtSUS5-S polypeptide sequence

MASTVADSMPDALKQSRYHMKRCFARFIAMGRRLMKLKHLTEEIEETIEDKAERTRILEGSLGKIMSSTQEAAVVPPYVAFAVRHNPGFWDYVKVNAETLSVEAI SAREYLKFKEMIFDEDWAKDDNALEVDFGAFDYSNPRLALSSSVGNGLNFISKVLSSKFGGKPEDAQPLLDYLLALNHQGENLMINENLNGVAKLQAALIVAEVF VSSFPKDTPYKDFEHKLKEWGFDKGWGHNAGRVRETMRLLSEIIQAPDPINMESFFSKLPTTTFNIVIFSIHGYFGQADVLGLPDTGGQVVYILDQVRALEEEMLQ RIKQQGLNVKPKILVVSRLIPDARGTTCNQEMEPILNSSHSHILRIPFRTEKGVLRQWDASAKILELMEGKPDLIIGNYTDGNLVASLLANKLGVTQGTIAHALE KTKYEDSDVKWKQFDPKYHFSCQFTADLLAMNAADFIITSTYQEIAGSETRPGQYESHTAFTMPGLYRAVSGINVFDPKFNIAAPGAEQSTYFPFTEKQKRFSTF RPAINELLYSNEENNEHIGFLADRKKPIIFSMARFDTVKNLSGLTEWYGKNKKLRNLVNLVIVGGFFDPSKSKDREEAAEIKKMHELIEKYQLKGQMRWIAAQTD KYRNSELYRTIADTKGAFVQPALYEAFGLTVIEAMDCGLPTFATNQGGPAEIIVDGVSGFHIDPYNGDESSKKIADFFEKCKVDSKYWNRISEGGLKRIEECYTW KIYANKVLNMGSIYGFWRQFNVGQKQAKQRYFEMFYNPLFRKLAKSVPIPHEEPLPLATSDSTQSQELKLPLPVPAAVAKVLPLTRHAFNLITSLPRVTGKVDVK

SEQ ID NO:28 : polynucleotide sequence of NtSUS5-T

atggcctcaactgttgctggtagcatgcctgatgctttgaaacaaagccgatatcatatgaagagatgcttcgctaggtgaacacccttcttgttctttttg ttttttccctctaccatttatgtcaaatttcaatgcataatgctaactactttttttctttttgacttcaaaattggacgtgaaaggttcattgcaatggga aggaggttgatgaagctgaaacatttaacagaagaaatagaaaaaactattgaagacaaggcagaaagaaccaagattttggagggttcacttggaaaaatt atgagttccacacaggtcagcaccatttaaccaacttaattgaataggaagaaaaaaaaaagcaaaagagttattgcaaggcgtaacgatttcctttgaaat tttcaggaggcagctgttgtcccaccttatgttgcttttgcagtaaggcacaatcctggcttctgggattatgtcaaagttgacgctgaaactctctctgtg gaagctatttcagccagggactatctcaaattcaaagagatgatctttgatgaagattggtaactggaagattgtatcattttaaagaaacaattttttaat attcaagattagttttgatggttgaatgtgcaagcagggcaaaggatgaaaatgcactcgaagtagattttggtgcttttgactactctaatcatcggttag ccctttcctcttctgtcggaaatgggctaaacttcatctcgaaagttttgtcttcaaagtttggtggaaaggcagaagatgcccagcctttgcttgattactt actagctcttaatcatcaaggagaggtatggaaatggactaccttcctttcttaaggaattatataatgatgtatgttataaagatcctttttaaacattga cactttgcagaatctaatgatcaatgagaatctgaatggcgtctctaagcttcaagcagcattgatagtagctgaagtttttgtatcttcctttcccaaaga cacaccttataaagactttgagcataagtaagcttttcaaacgcttctgttatcatatgcaatataccaagaatatgttgccttttgaaaagttgtttatgt ttatgacttgataatgaaaatactaggctcaaagaatggggctttgagaaagggtggggtcacaatgcaggaagagtaagagagacaatgagactgctttcc gagataatccaagcgccagatcccataaatatggagtcctttttcagcaggcttcctactacattcaacattgttatcttctccattcatggttactttggc caagcagatgtccttggtttgcccgatactggaggccaggtttacatacacagcaatttatctccttttgcctcatatttacttattagcgacacttgcatt attgaaatcacatttgtatttaacaggttgtttatattctggatcaagtaagagccttagagggaaatgttacaaagaatcaagcagcaagggttaaatg tgaagcccaagattcttgtggtgagttatgcaaaaatatgcgtagccaaggttttgaaattgttcagaggggattaagatgatcgagatatttgtttccttct tccattgatgtgtacaggtcactcgtctcattccagatgctcgagggactacatgcaatcaggagatggaacctatacttaactcgtcccattctcacatcctgagaattccattcaggacagagaaaggagttcttcgccaatgggtttctcggtttgatatctatccttacttggagaactatgccaaggcaagtctcctac caaaattaccacctattcatacactttattcagttttttgagctaatcattctcatttgtcacgtatgtgattaggatgcttctgctaagatacttgagctcatggaaggtaaaccagacctcattattgggaactacactgatggaaatttagtggcatctctattggccaacaaacttggagttactcaggttctacagctg atcatttatctgatcagattttctacattgttttcttgataattaaacggaaatcttatgagattgtaacattttagggaaccattgctcatgcattagagaaaaccaagtatgaagattctctgatgtcaagtggaagcagtttgattccaagtaccacttttcttgccaattcactgccgatttattggcaatgaatgctgctg attttatcattaccagcacatatcaagaaatcgcaggaaggttagcactgactctctcagtatatttggcaacttaatgaatgtactgcttgtggccaacactaaaagctattactcgtccttcagcgaaactaggcctggacaatatgaaagtcacacagcatttaccatgccggggctttatagagctgtttcaggcatcaat gtatttgatccaaagttcaacattgctgctcctggggctgaacagtctgcctatttccccttcactgagaaacagaaacgattcagcgcgtttcgtcctgct attgaggaactactttacagtaatgagcaaaacaacgagcacatgtaagtctaattgccccattttcctaatctaaccattgcttaaatgttctgtttttac ttgatatgtggtacttatcagtgatattttttattggaacagtggatttcttgcagaccgtaaaaaaccaattatattttcaatggcaagatttgatacggt gaagaacttgtcaggcttgactgagtggtatgggaagaataagaagttgcggaacttggttaacctcgttatcgttgggggattcttcgatccatcaaaatc aaaagaccgggaggaagcagctgaaatcaagaagatgcatgaattgattgagaaatacaagctcaagggacaaatgagatggatagcagctcaaactgataaatatcaaaacagtgagctatatcgaactattgctgacactaaaggagctttcgtccaaccggctttatatgaagcttttggactaactgttattgaagcaat gaattgtggactgcctacatttgctactaatcaaggcggacctgcagaaatcattgttgatggggtttcaggcttccatattgatccttacaatggggatga atcgagcaagaaaatagctgatttctttgagaagtgtaaggttgattctaaatattggaacaagatatgtggaggaggtctcaagcgcattgaagaatggtaa

SEQ ID NO:29 : NtSUS5-T polypeptide sequence

MASTVAGSMPDALKQSRYHMKRCFARFIAMGRRLMKLKHLTEEIEKTIEDKAERTKILEGSLGKIMSSTQEAAVVPPYVAFAVRHNPGFWDYVKVDAETLSVEAISARDYLKFKEMIFDEDWAKDENALEVDFGAFDYSNHRLALSSSVGNGLNFISKVLSSKFGGKAEDAQPLLDYLLALNH QGENLMINENLNGVSKLQAALIVAEVFVSSFPKDTPYKDFEHKLKEWGFEKGWGHNAGRVRETMRLLSEIIQAPDPINMESFFSRLPTTFNIVIFSIHGYFGQADVLGLPDTGGQVVYILDQVRALEEEMLQRIKQQGLNVKPKILVVTRLIPDARGTTCNQEMEPILNSSHSHILRIPFRTE KGVLRQWDASAKILELMEGKPDLIIGNYTDGNLVASLLANKLGVTQGTIAHALEKTKYEDSDVKWKQFDSKYHFSCQFTADLLAMNAADFIITSTYQEIAGSETRPGQYESHTAFTMPGLYRAVSGINVFDPKFNIAAPGAEQSAYFPFTEKQKRFSAFRPAIEELLYSNEQNNEHIGFLADR KKPIIFSMARFDTVKNLSGLTEWYGKNKKLRNLVNLVIVGGFFDPSKSKDREEAAEIKKMHELIEKYKLKGQMRWIAAQTDKYQNSELYRTIADTKGAFVQPALYEAFGLTVIEAMNCGLPTFATNQGGPAEIIVDGVSGFHIDPYNGDESSKKIADFFEKCKVDSKYWNKICGGGLKRIEEW

SEQ ID NO:30 : polynucleotide sequence of NtSUS6-S

atggctactgcaccagccctaaatagatcagagtccatagctgatagcatgccagaggccttaaggcaaagccggtaccacatgaagaaatgttttgccaagtacatagagcaaggaaagagg atgatgaaacttcataacttgatggatgagttggagaaagtaattgatgatcctgctgaaaggaaccatgttttggaaggcttacttggctacatattatgcactacaatggtatagctagat tcatatgtacttatgatgcccttatattgtttcctgatgtattactcttaaaaccttctttgatcaaatttacaggaggctgcagttgttcctccctacattgcctttgccacgagacagaat cctggattctgggaatatgtgaaagtgaatgctaatgatctttctgttgagggtattacagctacagaatacttgaaattcaaggaaatgatagttgatgaatgctggtatagtatacgttgc agcttatcataccttttgtggttttataacttcaatcagaaaactcatcagagttacctttgtgtgaacatgaaatgcagggcaaaagatgaatatgcactggaaattgattttggagcagta gacttctcaacgcctcgactgaccctatcctcttcaattggcaatggtctcagttatgtttccaagtttctaacttcaaagctaaatgctacctccgcgagtgcacagtgtctggttgactac ttgctcactttgaatcatcaaggagatgtacgtcaacaaaaatcaaactccataagtaaacttgtcaactctaagaagaaaaaataggaaaagaagattcacgtaacaaattttctttatgtt caactgcagaaactgatgatcaatgagacactcagcactgtctcaaagcttcaggctgcactggttgtagcagaagcatctatttcctctttaccaacagatacaccatatgagagctttgag ctaaggtgatttgttttttcctctacttccctccacttgtgccatgctacgtagtactaagtaacttcaattcttgtaaagattcaaacagtggggttttgagaaaggatggggtgatacagc tgaaagggtcagcgacaccatgagaacactgtctgaggtgcttcaggcaccagatccattgaacattcagaagttctttggaagggttccaactgttttcaatattgtattgttctctgtcca tggatactttggccaagcagatgttcttggcttgccagacactggtggtcaggtaagcatttaatagcttttacatttaacttctatgcattgacaataaaataatttttaacagtttgacca cttctgctcttgttcaacaggtagtttatgttttggatcaagttgtagcttttgaagaagaaatgctacaaagaattaaacagcaggggctcaatattaagcctcaaattcttgtggtgagtt cctagacaatcgacgtgactatgcaattatgtagaggctgtttagaaaagttaatatcatatgttgattgcacagttaacccgactgattccggatgcaaaaggaacaaagtgcaaccaggaa ctagaaccaatcaagaatacaaaacattcacacatcctcagagttccatttaggacagaaaaaggagtgcttaatcaatgggtttcacgatttgatatctctatccatatctggagagatatact caggtatgtatttttatatcaaccttgctcatcaaagatgtgttgtttcctcaattccatttttccccttggcaaaaggatgctgctgacaaaatcgtcgagctaatggaaggcaaacctgat ctaatcattggtaactacactgatgggaatctagtggcttcactaatggctagaaaacttgggataactctggtaacttttcttaatcatatttgatgttgcttcttctccaagttagttctt aatctccactgacctagaccatctttgcaacagggaactattgctcatgctttggagaagacaaaatatgaagactctctgacataaaattgaaggaactcgatccgaagtaccacttctcttgc caattcacagctgatttgattgcaatgaattcagcagatttcattatcactagcacataccaagaaatagctggaaggtaagaattagagctaataagtaatgcattcatatgtatttcagca tcgctctttcaccatcatcgaatacacaccactactcagtaaatgtatttgctcaaaagtttgcaacttaatggatctcattcttgaatgcttcaacatatgcagcaaagataaaccaggaca gtatgagagccatagtgcatttacccttccagggctttacagagttgcttcaggtatcaatgtctttgatccaaaatttaatattgctgcacctggggcagaccagtcggtgtatttccctta cacagaaaagcagaagcgtttgactgctttccgccctgccattgaggaactgctttttagtaaagtggacaatgacgagcacgtgtaagtctaagtgttaaacttcagcttagtgcctagaac atcccactgctctatgtattgatgtttcacttgtttcaaacagtggatatttagaagacagaaagaaacctatcctgtttaccatggcaaggctggacacagtgaagaacacatctggactaa cagaatggtatggcaagaacaagaggctcagaagcttagttaaccttgttgtggttggtggttcctttgatcctacaaaatccaaggatagggaagaagcagctgaaataaaaaagatgcaca tgctgatagagaaataccagcttaagggtcagattagatggatagcagctcagactgacagatacagaaatagtgaactctaccgcacaatagcagattccaaaggagcttttgtgcagcctg cattgtatgaagcatttggtctaacagtcattgaggcaatgaactgtggattaccaacctttgctaccaaccaaggtggccctgctgagattattgttgatggggtctcaggctttcatattg atccaaataatggggatgaatcaagcaacaaaattgccaactttttccaaaaatgcagggaggatcctgagtattggaacaggatttcagtccagggtctaaaccgtatatatgaatggtaac tcacagataagccattcaaattgcaaagaggcacatatcttgcagaaaatttcttaatccttaaatcctaattttttgcagttacacatggaagatctatgcaaacaaggtattgaatatggg gtccatctatactttttggaggacattgtacagagatcagaaacaagcaaagcaaagatacatcgagactttctacaatcttgagtttaggaacttggtatagtgctgcatgacattgacagt ataccacaaacatctttatgagatgaattacttttaataaaattgtttttaacctttgcttccttaatggcacttattgcaggtaaaaaatgtgcctatcagaaaggacgaaacaccacaagg accaaaggagaggggagaaagttaagccacagatatcacaaaggcatgctctaaagcttttgcctacagtttttcaagagaccctagtatattctagtactaaattagaattatacagcatgca gcttttgctgttcacctttctaaatcaccagttgtgtcaatcaagttgacaaaatcaataaattgggattttccctttcctatgcttgattgttattactcctactttgtttatggtagtctt ccttcattgttttctcctgtacttcttttactacaactgtactgacatactaattatttctgtgtaccaggcgctcacaatcaaggttgcagaagtaagattagataaaattgctactgcatga

SEQ ID NO:31 : NtSUS6-S polypeptide sequence

MATAPALNRSESIADSMPEALRQSRYHMKKCFAKYIEQGKRMMKLHNLMDELEKVIDDPAERNHVLEGLLGYILCTTMEAAVVPPYIAFATRQNPGFWEYVKVNAND LSVEGITATEYLKFKEMIVDECWAKDEYALEIDFGAVDFSTPRLTLSSSIGNGLSYVSKFLTSKLNATSASAQCLVDYLLTLNHQGDKLMINETLSTVSKLQAALVVA EASISSLPTDTPYESFELRFKQWGFEKGWGDTAERVSDTMRTLSEVLQAPDPLNIQKFFGRVPTVFNIVLFSVHGYFGQADVLGLPDTGGQVVYVLDQVVAFEEEML QRIKQQGLNIKPQILVLTRLIPDAKGTKCNQELEPIKNTKHSHILRVPFRTEKGVLNQWVSRFDIYPYLERYTQDAADKIVELMEGKPDLIIGNYTDGNLVASLMARK LGITLGTIAHALEKTKYEDSDIKLKELDPKYHFSCQFTADLIAMNSADFIITSTYQEIAGSKDKPGQYESHSAFTLPGLYRVASGINVFDPKFNIAAPGADQSVYFP YTEKQKRLTAFRPAIEELLFSKVDNDEHVGYLEDRKKPILFTMARLDTVKNTSGLTEWYGKNKRLRSLVNLVVVGGSFDPTKSKDREEAAEIKKMHMLIEKYQLKGQI RWIAAQTDRYRNSELYRTIADSKGAFVQPALYEAFGLTVIEAMNCGLPTFATNQGGPAEIIVDGVSGFHIDPNNGDESSNKIANFFQKCREDPEYWNRISVQGLNRI YECYTWKIYANKVLNMGSIYTFWRTLYRDQKQAKQRYIETFYNLEFRNLVKNVPIRKDETPQGPKEREKVKPQISQRHALKLLPTVFQETLALTIKVAEVRLDKIATA

SEQ ID NO:32 : polynucleotide sequence of NtSUS6-T

atggctactgcaccagccctgaaaagatcagagtccatagctgatagcatgccagaggccttaaggcaaagccggtaccacatgaagaaatgttttgccaagtacatagagcaaggcaagag gatgatgaaacttcataacttgatggatgaattggagaaagtaattgatgatcctgctgaaaggaaccatgttttggaaggcttacttggctacatattatgtactacaatggtatagctag attcatatgtacttatgatgtccttatattgtttccggaggcattattcttaaatccttctttgatcaaatttgtaggaggctgcagttgttcctccctatattgccttcgccacgagacag aatcctggattctgggaatatgtgaaagtcaatgctaatgatctttctgttgagggtattacagctacagattacttgaaattcaaggaaatgatagttgatgaaagctggtatagaatact ttgcagcttatcataccttttgtggttttataatttcaatcagaaaactcatcagagttacctttgtgtgaacatgacatgcagggcaaaagatgaatatgcactggaaattgattttggag cagtagacttctcaacgcctcgactgaccctatcctcttcaattggaaatggtctcagttatgtttccaagtttctaacttcaaagctaaatgctacctcagcgagtgcacagtgtctggtt gactacttgctcactttgaatcaccaaggagatgtacgtcaacaaaaatcaaactccataagtaaacttgtcaactctaagaagtaaaaataggaaaagaagattcatgtaacaaattttct ttatgttcaactgtagaaactgatgatcaatgagacactcggcactgtctcaaagcttcaggctgcactggttgtagcagaagcatctatttcctccttaccaacagatacaccataccaga gctttgagctaaggtgatttgttttttcctctacttccttccacttttggtgtgctacatagtactaagtaacttcaattcttgtaaagattcaaacagtggggttttgagaaaggatgggg tgatacagctgaaagggtccgcgacaccatgagaacactttctgaggtacttcaggcgccagatccattgaacattgagaagttctttgggaggttccaactgttttcaatattgtattgt tctctgttcatggatactttggccaagcaaatgttcttggcttgccagacacaggtggtcaggtaagcatctaatagcttttacatttaacttctatgcattgacaataaaataacttctac actaccaaataatttttgaaagtttgaccacttcggctcttgttcaacaggtggtttatgttttggatcaagttgtagcttttgaagaagaaatgctccaaagaattaaacagcaggggctc aatattaagcctcaaattcttgtggtgagctcctagacaatgacgtgactatgcaattaagtagaggctgtttagaaaagttaatatcatatgttgattgcacagttaacccgactgattcc ggacgccaaaggaacaaagtgcaaccaggaactagaaccaatcaagaatacaaaacattcacacatcctcagagttccatttaggacagaaaaaggagtgcttaatcaatgggtttcacgat ttgatatctatccatatctggagagatactcaggtgtgtatttttatatcaaccctgctcatcaaagatgtgttgtttcctcaattccattttcgccttgacaaaaggacgctgctgac aaaatcatcgagctaatggaaggcaaacctgatctaatcattggtaactacactgatgggaatctagtggcttctctaatggctagaaagcttgggataactctctggtaacttttcttatcat atttgatgttgtttcttctccaagttggttcttaatgtcaactaacccagaccatctttgtaacaggaactattgctcatgctctggagaagacaaaatatgaagactctgacatcaaatt gaaggaactcgatccgaagtaccacttttcttgccaattcacagctgatttgattgcaatgaattcagcagatttcattatcacaagcacatatcaagaaatagccggaaggtaagaattgg aactacggaagcagagagctaataagtagtgcactcatatatttcagcatcgctctttcgcataatcgaatacacaccactcagtaaatgtacttgctcaaaagtttacaagtttatgg atcttattcttgaatgcttcaacatatgcagcaaagataggccaggacagtatgagagccatagtgcatttacccttccagggctttacagagttgcttcaggcatcaatgtctttgatcct aaatttaatattgctgcacctggggcagaccaatcggtgtatttcccttacacagaaaagcagacgcgtttgactgctttccgccctgccattgaggaactgctttttagtaaagtggacaa tgacgagcacatgtaagtcttagtgttaaacttcagctttcagcttagtgcctagaacattccactggctctatgtattaatgtttcacttgtttcaaacacagtggatatttagaagacag aaagaaacctatcctgtttaccatggcaaggctggacacagtgaagaacacatctggactaacagaatggtatggcaagaacaagaggctcagaagcttagttaaccttgttgtggttggtg gttcctttgatcctacaaaatccaaggatagagaagaagcagctgaaataaaaaagatgcacatgctgatagagaaataccagcttaagggtcagatcagatggatagcagctcagactgac agatatagaaacagtgaactctaccgcacaatagcagattccaaaggagcttttgtgcagcctgcattatatgaagcatttggtctaacagtcattgaggcaatgaactgtggattaccaac ctttgctaccaaccaaggtggccctgctgagattattgttgatggggtctcaggctttcatattgatccaaataatggggatgaatcaagcaacaaagttgccaactttttccaaaaatgca gggaggatcctgagtattggaacaggatttcagtccagggtctaaaccgtatatgaatggtaactcacagataagccattcaaattgcaaagaggcacatatctcttgctgaaaatttctta atcctttaatcctaaaattttgcagttacacatggaagatctatgcaaacaaggtattgaatatggggtccatctatactttttggaggacattgtacagagatcagaaacaagcaaagcaa agatacatcgagactttctacaatcttgagtttaggaacttggtatagtgctgcatgacattgacagtataccacaaacatctttatgagatgaattacttttaataaaattgtttttaacc tttgcctccttaatgacacttattgcaggtaaaaaatgtgcctatcagacaggacgaaacaccacaaggaccaaaggagaggagggagaaagttaagccacagatatcacaaaggcatgctc taaagcttttgcctatagtttttcaggagaccctagtatattctagtactaaattagaattatacagcatgcagcttgcttctgctgttcacctttctaaatcaccagttatgtcaatcaag ttgacaaaatcaataaattcggcttttccctttcctatgcttgattgttattactcctacttcgtttatggtagtcttccttcattgttttctcctgtacttcttttactacaactgtactga

SEQ ID NO:33 : NtSUS6-T polypeptide sequence

MATAPALKRSESIADSMPEALRQSRYHMKKCFAKYIEQGKRMMKLHNLMDELEKVIDDPAERNHVLEGLLGYILCTTMEAAVVPPYIAFATRQNPGFWEYVKVNANDLSVEGIT ATDYLKFKEMIVDESWAKDEYALEIDFGAVDFSTPRLTLSSSIGNGLSYVSKFLTSKLNATSASAQCLVDYLLTLNHQGDKLMINETLGTVSKLQAALVVAEASISSLPTDTPY QSFELRFKQWGFEKGWGDTAERVRDTMRTLSEVLQAPDPLNIEKFFGRVPTVFNIVLFSVHGYFGQANVLGLPDTGGQVVYVLDQVVAFEEEMLQRIKQQGLNIKPQILVLTRL IPDAKGTKCNQELEPIKNTKHSHILRVPFRTEKGVLNQWVSRFDIYPYLERYTQDAADKIIELMEGKPDLIIGNYTDGNLVASLMARKLGITLGTIAHALEKTKYEDSDIKLKE LDPKYHFSCQFTADLIAMNSADFIITSTYQEIAGSKDRPGQYESHSAFTLPGLYRVASGINVFDPKFNIAAPGADQSVYFPYTEKQTRLTAFRPAIEELLFSKVDNDEHIGYLE DRKKPILFTMARLDTVKNTSGLTEWYGKNKRLRSLVNLVVVGGSFDPTKSKDREEAAEIKKMHMLIEKYQLKGQIRWIAAQTDRYRNSELYRTIADSKGAFVQPALYEAFGLTV IEAMNCGLPTFATNQGGPAEIIVDGVSGFHIDPNNGDESSNKVANFFQKCREDPEYWNRISVQGLNRIYECYTWKIYANKVLNMGSIYTFWRTLYRDQKQAKQRYIETFYNLEF RNLVKNVPIRQDETPQGPKERREKVKPQISQRHALKLLPIVFQETLVYSSTKLELYSMQLASAVHLSKSPVMSIKLTKSINSAFPFPMLDCYYSYFVYGSLPSLFSPVLLLLQLY

--->

List of sequences

<110> Philip Morris Products S.A.

<120> MODULATION OF REDUCING SUGARS CONTENT IN PLANT (INV)

<130> P10682EP

<160> 33

<170> PatentIn version 3.5

<210> 1

<211> 4237

<212> DNA

<213> Nicotiana tabacum

<400> 1

atggcggaaa caaacaatag cgttccttac acccaattac cggcggagga caataacacc 60

tccgttaatt ctccggccgg atgccggcta cgacccaaaa gagtgtcgtt tatagtatta 120

acagggctgg tggcagcttt gttacttttt gtggcagtga aatatgggaa aaacgaggcg 180

gaggatgtga atccagggcc agtaccacca caagaaaccg tgtgcaatat gcttggttct 240

aatctaatgc cgctgaccag catgaagacg gtggcgcgtg gggtggcaga aggtgtctcc 300

gccaagtcac gcggtcgttt cttgggatta cggccgtttc catggaccaa acaaatgttg 360

gcttggcaaa gaacatcctt ccactttcaa cctaagaaga attggatgaa tggttagtaa 420

ttctttttct cttatgttat taattttcat aaatcaactt tattattatt attatacaat 480

aaatcaacat tgcttattga tgaattttaa cataaacccg ccttatgctt gacgagatta 540

actagaacta tatatacaat gaatgattat ctccattcca ttacataacc atgaattatg 600

tttcttaatt aattaaagat ttgacatgac attatatttc gtttatagtt taagaaaagc 660

tttgtattga tgtaaaagaa accattacag cttcgaatat gggatacctt gtctttttct 720

tttcctaaga tggatctttg attgcaagaa cagagtttga attactcagg aaacttattt 780

gcttatttat tattttttga ggtgaacatt aatgatttat tcttatttgg catgtgttgg 840

attatttggc ttggattgcg ctgatcacgg aaattgcctg attcttttcg tcagatccta 900

atggtaaagt ccatatattt ctacttgtta ttgttgttgt tcttcttatt attatattat 960

tattgaaaat tatcgacata atcgggacct caaaacatac tagtcgtagc agtttttaag 1020

tagacagatt gtcaatatga tgaagacagt tgttttcaga caattgcatg tgaattttct 1080

aggagcaaac acaaattcct agaatggtaa gcaacttcca ccctgtctgt tccaattata 1140

acctcgctac ttttgatcca cttaatctta ttcaaccaac agtggatcac ttatttaatt 1200

atatgtgacc tagtttattg agacattttt acattaagcc ctttcgtatt tacacttcaa 1260

tatgcatcat acaaaaaaaa atgtacttca aagttatacg ttatattaat ttctaactcc 1320

aatttttaaa aaaaaatata ttttaggtcc cttattctac aaaggatggt accatttgtt 1380

ctatcaatac aatccagagg ctgcagtatg gggaaatatt gtatggggac atgcagtttc 1440

aagagactta attcactggc aacaccttcc agttgctatg gttgcggatc aatggtacga 1500

cattaatggt gtatggaccg gatccgcaac cattttaccc gatggtaaac tcgtcatgtt 1560

gtatactggg tcaaccaacg agtcagtaca ggttcaaaat ttagcgtacc cggctgaccc 1620

atcggatcct ctcctaataa aatgggtcaa gtatgagggc aacccggttc ttgtaccacc 1680

acccggaatt gctgctaagg atttccgtga ccccaccact gcatggacca caccacaagg 1740

caaatggcgg attactattg gttcaaaagt taataaaact ggaatttcat tggtctatga 1800

cactattgat tttaagaatt ttgagttgct ggatggggtg ctccatggtg tatcgggtac 1860

gggtatgtgg gaatgtgtgg atttttaccc ggtttcgaaa gttgttgaaa atgggcttga 1920

cacttcagat aatgggcctg cagtaaaaca tgtgttaaag tccagtcttg atgatgatag 1980

aaatgattat tatgcacttg gaacttatga tgctgtggct ggaaaatggg ttcctgataa 2040

tcccactatt gatgttggta ttggattaag atatgattat ggaaattttt atgcatcaaa 2100

aacattttat gaccaagaga aaaagagaag agtcctttgg gcttggatta ctgaaagtga 2160

tagtgaagct gctgatattt gcaaaggttg ggcatcactt caggtacaat tcaattgtgt 2220

caagctagcg cttgcacata gatttagttg aaacctaaaa aatgagtatt tgaaattccg 2280

tagaaaaata atttttgaaa gttgaagttg tgtttgaata tgcattttat ttgaaaaaaa 2340

aaacagttct aattttatga gaaagaaaaa ttcacctaaa aactgcccta aaccagattt 2400

taggaacttg aaaaaaaaat aaactttttc aaaaactgat tatattctat gaacaaacaa 2460

tattatcaaa aatctatttt ttttttgcca aaatctatgg ccaaacagga gctaatttcc 2520

tttatttttt ttttcaaact tcatgtcata tttgaatttt ggtctcattt aacactttgg 2580

taacgtgtga tgtaacacag cccattccaa ggactataaa atatgacaag aagacaggaa 2640

gcaatataat tacttggcca gtggcagagg ttgagaattt gagatttaac agcaaggaat 2700

tcgacaaggt ggaggtcaag ccadgaaatg ttgttccact agaagttggc actgccactc 2760

aggtttgttc attaaattta gcttatatac actgactgcc taaaagaatt tttttgacat 2820

tattagtgta ttttaagcta ttatagcacg taacatgcta atgctcgaat aagtttaact 2880

tactataact tgaattgttg atgattacag ttggacataa tggctgagtt tgaagtagac 2940

cctaaggtct tggagaaatt agaaggaagt aatgctacat atgagtgcag aagcagcggt 3000

ggatctgctg aacgtggtgc cttaggacca tttggtttat tggttttaac agataaggga 3060

ttgtccgagc aaactccaat ttacttctac attgctaaag acgctgctgg aaatttcacc 3120

acattcttct gcaatgatct taccaggttc taatttctcc tctcttgcat tttcatctca 3180

tcaatgaagt tttagccctt caccccctcc cccaaaacca aactaataaa ttggagaaaa 3240

ccctttattg gttcagtgct taatagcagt acggaattca ggattttaag tcagtgggtt 3300

ctgcgatcta tatatatata ataatatttt tctgcacata catatagtcc gagctagaca 3360

tagtgagttc cgttgaacct gttgcattta gtctgagtcc gccactgctt aagcacatcc 3420

ttctcataac aaccgagctt tccaaaaact taagtatttc tcatgtccat acttttattc 3480

atgtttgaaa atgaagtcac attttgtttt ataaccgaaa aatcccgagg gcaagtggcc 3540

agtacatggt tcgaagctca atggacactg gcaccgcccc tttatcgtgc tccacttaaa 3600

tactaagatt ttgtccgtgg cagggtttca accaatcacg tacgtttaac tcatatatta 3660

ggaatagctt ttaccactag accaaaactc ggggacaatg tatgaagccg gatatttgtt 3720

gcaattcttt ttaaattaaa atggggacaa gatccgagac aaatcttgaa aatgcattac 3780

gaagtattgt taagtaagta tgaaaatggt gattctcatc tttttacttc cttttttagg 3840

tcatctgaag caacagatgt tcgcaaacta atctacggaa gcacagttcc agtcctccaa 3900

ggagagaagc tttctctaag aacactggta atatcccctt tttctttctt aatttcttaa 3960

tccaaattct taattagtgc ttgttttcct ttgtgcgtat aattaagttt actaagtatc 4020

aattaatggg gtatttttgt caatgtaata ggtggatcat tcaatagtag aaagttttgc 4080

acaaaatgga aggacagcaa taacatcaag gttatatcca acaaaggcaa tatatgaaga 4140

tgctaagctc tacttgttta acaatgctac agatgttacc attactgcct cggtcaagat 4200

ttggcaaata cattctgcaa atatacaatc tagttaa 4237

<210> 2

<211> 649

<212> PROTEIN

<213> Nicotiana tabacum

<400> 2

Met Ala Glu Thr Asn Asn Ser Val Pro Tyr Thr Gln Leu Pro Ala Glu

1 5 10 15

Asp Asn Asn Thr Ser Val Asn Ser Pro Ala Gly Cys Arg Leu Arg Pro

20 25 30

Lys Arg Val Ser Phe Ile Val Leu Thr Gly Leu Val Ala Ala Leu Leu

35 40 45

Leu Phe Val Ala Val Lys Tyr Gly Lys Asn Glu Ala Glu Asp Val Asn

50 55 60

Pro Gly Pro Val Pro Pro Gln Glu Thr Val Cys Asn Met Leu Gly Ser

65 70 75 80

Asn Leu Met Pro Leu Thr Ser Met Lys Thr Val Ala Arg Gly Val Ala

85 90 95

Glu Gly Val Ser Ala Lys Ser Arg Gly Arg Phe Leu Gly Leu Arg Pro

100 105 110

Phe Pro Trp Thr Lys Gln Met Leu Ala Trp Gln Arg Thr Ser Phe His

115 120 125

Phe Gln Pro Lys Lys Asn Trp Met Asn Asp Pro Asn Gly Pro Leu Phe

130 135 140

Tyr Lys Gly Trp Tyr His Leu Phe Tyr Gln Tyr Asn Pro Glu Ala Ala

145 150 155 160

Val Trp Gly Asn Ile Val Trp Gly His Ala Val Ser Arg Asp Leu Ile

165 170 175

His Trp Gln His Leu Pro Val Ala Met Val Ala Asp Gln Trp Tyr Asp

180 185 190

Ile Asn Gly Val Trp Thr Gly Ser Ala Thr Ile Leu Pro Asp Gly Lys

195 200 205

Leu Val Met Leu Tyr Thr Gly Ser Thr Asn Glu Ser Val Gln Val Gln

210 215 220

Asn Leu Ala Tyr Pro Ala Asp Pro Ser Asp Pro Leu Leu Ile Lys Trp

225 230 235 240

Val Lys Tyr Glu Gly Asn Pro Val Leu Val Pro Pro Pro Gly Ile Ala

245 250 255

Ala Lys Asp Phe Arg Asp Pro Thr Thr Ala Trp Thr Thr Pro Gln Gly

260 265 270

Lys Trp Arg Ile Thr Ile Gly Ser Lys Val Asn Lys Thr Gly Ile Ser

275 280 285

Leu Val Tyr Asp Thr Ile Asp Phe Lys Asn Phe Glu Leu Leu Asp Gly

290 295 300

Val Leu His Gly Val Ser Gly Thr Gly Met Trp Glu Cys Val Asp Phe

305 310 315 320

Tyr Pro Val Ser Lys Val Val Glu Asn Gly Leu Asp Thr Ser Asp Asn

325 330 335

Gly Pro Ala Val Lys His Val Leu Lys Ser Ser Leu Asp Asp Asp Arg

340 345 350

Asn Asp Tyr Tyr Ala Leu Gly Thr Tyr Asp Ala Val Ala Gly Lys Trp

355 360 365

Val Pro Asp Asn Pro Thr Ile Asp Val Gly Ile Gly Leu Arg Tyr Asp

370 375 380

Tyr Gly Asn Phe Tyr Ala Ser Lys Thr Phe Tyr Asp Gln Glu Lys Lys

385 390 395 400

Arg Arg Val Leu Trp Ala Trp Ile Thr Glu Ser Asp Ser Glu Ala Ala

405 410 415

Asp Ile Cys Lys Gly Trp Ala Ser Leu Gln Pro Ile Pro Arg Thr Ile

420 425 430

Lys Tyr Asp Lys Lys Thr Gly Ser Asn Ile Ile Thr Trp Pro Val Ala

435 440 445

Glu Val Glu Asn Leu Arg Phe Asn Ser Lys Glu Phe Asp Lys Val Glu

450 455 460

Val Lys Pro Gly Asn Val Val Pro Leu Glu Val Gly Thr Ala Thr Gln

465 470 475 480

Leu Asp Ile Met Ala Glu Phe Glu Val Asp Pro Lys Val Leu Glu Lys

485 490 495

Leu Glu Gly Ser Asn Ala Thr Tyr Glu Cys Arg Ser Ser Gly Gly Ser

500 505 510

Ala Glu Arg Gly Ala Leu Gly Pro Phe Gly Leu Leu Val Leu Thr Asp

515 520 525

Lys Gly Leu Ser Glu Gln Thr Pro Ile Tyr Phe Tyr Ile Ala Lys Asp

530 535 540

Ala Ala Gly Asn Phe Thr Thr Phe Phe Cys Asn Asp Leu Thr Arg Ser

545 550 555 560

Ser Glu Ala Thr Asp Val Arg Lys Leu Ile Tyr Gly Ser Thr Val Pro

565 570 575

Val Leu Gln Gly Glu Lys Leu Ser Leu Arg Thr Leu Val Asp His Ser

580 585 590

Ile Val Glu Ser Phe Ala Gln Asn Gly Arg Thr Ala Ile Thr Ser Arg

595 600 605

Leu Tyr Pro Thr Lys Ala Ile Tyr Glu Asp Ala Lys Leu Tyr Leu Phe

610 615 620

Asn Asn Ala Thr Asp Val Thr Ile Thr Ala Ser Val Lys Ile Trp Gln

625 630 635 640

Ile His Ser Ala Asn Ile Gln Ser Ser

645

<210> 3

<211> 5883

<212> DNA

<213> Nicotiana tabacum

<400> 3

atggcggaaa caaacaatag cgttccttac acccaattac cggcggagga caataacacc 60

tccagtaatt ctccggccaa atgccggcga cgacccaaaa gagtgtcgtt catagtatta 120

acagggctgg tggcagcttt gttacttttt gtggcagtga aatatgggaa taacgaggcg 180

gaggatgtaa atccaggggcc agtaccacca caagaaaccg tgtgtaacat gcttggttct 240

aatctaatgc cgctgaccac catgaggacg gtggcgcgtg gggtggcaga aggtgtctcc 300

gccaagtcac gcggtcgttt cttgggatta cggccgtttc catggaccaa acaaatgttg 360

gcttggcaaa gaacatcctt ccactttcaa cctaagaaga attggatgaa tggttagtaa 420

ttctttttct cttatgttat tttcataaat cagctttgtt tttattaaac aataaatcaa 480

cagcttattg ataattttaa acataaaacc gccttatgct tgacgagatt aactagaact 540

ttatgtacaa tgaatggtta tctccattcc attacatgcc catgaatttt atgtgtctta 600

atttaaagat ttgacaggac attacattac gtttatagtt taagaaaagc ttggtattga 660

tataaaaaaa accattacag cttcgaatat gggatacctt gtctttttct ttgcctaaga 720

tggatctttg attgcaagaa cagagtttga attactcagg aaaaatatga aatcgttttg 780

gaacttattt gcttgtttat tattttttga ggtgaacatt aatgatttat tcttatttgg 840

catgtgttgg attctttggc tttggactgc gttgctcacg gaaattacct gattctgttc 900

gtcagatcct aatggtaagt ccatattttc tgccggtatt attattatta ttattgttat 960

tgttattatt attattatta ttattaattt attttgatat attggaaacc atcgacaaaa 1020

cggggacctc aaaacatact agtcggggta gtttgtaagt agacagattg acaatatgat 1080

gaagacagtt gtctttagac aattgcatgt gaattttgta ggagcaaaca caaattccta 1140

gaatggtata caacttcaat cctgtctgtc caattataac ctcgctactt ttgatccact 1200

acaccttttt cgatcaacag gggatcactt atttaattat acggaaccct ttatataaca 1260

atcatatttg ttcccgtatt ttttaggttt tatattgagt ggttgttatg caaatattac 1320

aggatttgac gtttaaatat ttttttggtt gttatagata aaaattatct ataaataaat 1380

aatcattcct tttttcatgt tacatataaa aaataaggaa attatttaaa tttaaaatct 1440

cacaagctat gcatatttca ctaattaaat attaaagaaa gttaatacat tattaataaa 1500

ttcataacta aaaatataaa gatttaaact ctaaggcaga cattttaagg ctaccaaaaa 1560

aataattttt ttcaagattg atggaagtgg aacaactttg taattgtagc ttgtttcgta 1620

gatttcattc tcttactagc gatataacac ttgaattggc aaagatccgt gtctaaattt 1680

tcagcaaaaa ggtatccaat agtttttcct tgaatacaat ctaagagttt aaccgttaaa 1740

ttttctatcc aagtattttt taaatttatc caataaaaaa gatatcatgt gcaaatctat 1800

aaatcttata ttttatgcaa gatagaaact ttatttattt ttagaattat tattagcaat 1860

cttaaagatt ttatatggct gttatagagg ggtaatttta caaaaagcgt tctgctataa 1920

atatggttgt tgctgttata ggtaaaaagt tgttataaaa ttgtttatga aagtcacttt 1980

actttatttt ttaactgaaa agtcactata ctttgcacat tgtaactcaa aagtcaatca 2040

acacttttag ggcgttatta aacattattt ttcatatttc ttttcagccc aaccatttaa 2100

aaaataaaaa aaaatattta aatcatgact cgacggttcc atgacctgac ccattttctc 2160

ttatttatgt tataaaaaac ataaatatta ctttgttgta gtgtattata ttggttctca 2220

atatagttat actaaatgta tacattggtt taagcaaagt attaaataga gaataaggaa 2280

ttaatcttaa caaactagag gagttagatt tgaaactgaa acaagaaaat tagtagcgtt 2340

gaagtgaaaa aaaataaaaa ggaggaagaa aaaataaaaa aagtatttat gcttgaaatt 2400

ttgactgaga aatattaata taagagtaaa ttaaaagaca atatccttga tgttttagaa 2460

caagaacgcg cattttagta aagataatgt tactagacga ccatttttag tcgaactaat 2520

ttaatacttt gcttaaatca atatgttgaa aatcaagaca attaacagta aagtaatatt 2580

tacgtttttt tgtaacaaaa ataagagagt gggtcgagtt aaagagcggg ttgagtcacg 2640

gtttaaatgg gtattttttt attgtttaaa tgattgagtt aaaaaaaata tgaaaaaaaa 2700

aatttaatat ggcctaaatg tattgagtga cttttgagtt acaatgtgta aagtatgatg 2760

actttccttt tacaaaacaa agtaaagtga ctttcataaa caattttcat tagttcaatg 2820

acttctgaga aatggactcc ttaaaaaatt gattctgaag aaaacttggt ttttacggtg 2880

aatgactgtt atatatgaat gttgttatcg aaaggtctga ctgtatgtga cctagtttat 2940

tgagacgttt ttacattaaa gccctttcgt atttacactt caatatgcat catacaaaaa 3000

atatgtgctt cataattata cattacattc atttctaact ccacttttac aaaaaatatt 3060

ttaggtccat tattctacaa aggatggtac catttgttct atcaatacaa tccagaggct 3120

gcagtatggg gaaatattgt atggggccat gcagtttcaa gagacttaat tcactggcaa 3180

caccttccag ttgctatggt tgcggatcaa tggtacgaca ttaacggtgt atggaccgga 3240

tccgcaacca ttttacccga tggtaaactc gtcatgttat ataccgggtc aaccaacgag 3300

tcagtacagg ttcaaaatct agcgtacccg gctgacccat cggatcctct cctaagaaaa 3360

tgggtcaaat atgagggcaa cccggtactt gtaccaccac ccggaattgc tactaaagat 3420

tttcgtgacc ccaccactgc atggaccaca ccacaaggca aatggaggat tactattggt 3480

tcaaaggtta ataaaactgg aatttcattg gtctatgaca ctattgattt taagaaattt 3540

gagttgttgg atggggtgct ccatggtgta ccgggtacgg gtatgtggga atgtgtggac 3600

ttttacccgg tttcgaaagt tgttgaaaat gggcttgaca catcagataa tgggcctgca 3660

gtaaaacatg tgttaaagtc cagtctagat gatgatagaa atgattatta tgcacttgga 3720

acttatgatg cagtggctgg taaatggatt cctgataatc ccacaattga tgttggtatt 3780

ggattaagat atgattatgg aaatttttac gcatcaaaaa cattttatga ccaagaaaaa 3840

aagagaagag tcctttgggc ttggattact gaaggtgata gtgaagctgc tgatatttgc 3900

aaaggttggg catcacttca ggtacaattc aattgtgtcg aagacaattt agctagtgtt 3960

gggatataga tttggttgaa acttaaaaaa aaaaatattt aaaattatgg acatgtattt 4020

tatttgaaaa aaattaaaat tctgtgagtg gaagaaaacc ttttacccaa aaactaccct 4080

aaaccagatt ttgggaatgt aaaaaaaaga atcagatcat attctatgaa caaacaatat 4140

tatcaaaagt tttttaaaaa caattttcaa aatctatggt caatttcctc ttttatttta 4200

cttcattttg tcatatttga attttggtct catttaacac ttggtaacgt gtgatgtaaa 4260

acagcctatt ccaaggacta taaaatatga caagaagaca ggaagcaaca taattacttg 4320

gccagtggcg gaggttgaga atttgagatt aaacagtaag gaattcgaca aggtggaggt 4380

aaaaccaggg tcagtttttc cactagaagt tggcactgcc actcaggttt gttgattgaa 4440

tttaactata cacgtgtaaa agaatttctt tacgttatcg gtctatttta aactattata 4500

gcacgtaaca tgctaatatt cgataagttt aacttactat aatttgaatt gttgatgatt 4560

atagttggac ataatggctg agtttgaaat agaccctaag gtcttggaga gattagaagg 4620

aaataatgct acatatgagt gcagaagcag tgggggatct gctgaacgtg gtgccttagg 4680

accatttggt ttattggttt taacagataa gggcttgtcc gagcaaactc caatttactt 4740

ctacattgca aaagacgctg ctggaaattt caccacattc ttctgcaatg atcttaccag 4800

gttctaattt ctcctctctt gcattttcat ctcatcaatg aagttttagc ccctcccccc 4860

caccaaaacc aaactaagaa attggagaaa aacctttatt ggttcactgc ttaatagcag 4920

tacggaattc aggattttga gtcattaggt tctgctctat atatatatat ataataatat 4980

ttttctacac atatatatag ttcgagctaa acataatgag ttccgtcgaa cctgttgcat 5040

ctagtctgaa tccgccactg ctttaacaca tctttctcat aataaccact atttccaaga 5100

gcttaagtat ttctcatgtc catacttcta tccacgttta aaaatgaagt cagattttgt 5160

tttatatccg agaaatcccg agggcaagtg gccagtacat ggttcgaagc tcaatggaca 5220

ctggcaccgc cctttatcgt gctctactta aatattaaga ttttgtctgt tgcagggttt 5280

taaccaagga cgtacgttta acccatatat aacgagtagc ttttaccact agaccaaaac 5340

tcggggcaat atatgaagcc agatatttgt tgcaattctc tttaattaaa ttaaaatggt 5400

gacaagatcc gagacaaatc ttggaagtgc attacgtagt atttttaagt aagtatgata 5460

atggtgattc tcatcttttt acttcctttt ttttaggtca tctgaagcaa cagatgttcg 5520

caaactaatc tacggaagca cagttccagt cctccaagga gagaagcttt ctctaagaac 5580

actggtaatt ttcatttttc ttcttttttt aattgcttat tcaaaattct tgattatatt 5640

gcgtacactt aagtttacca aatataaatt aatggggtat ttttgtgaat gtaataggtg 5700

gatcattcaa tagtagaaag ttttgcacaa agtggaagga cagcaataac gtcaagggta 5760

tatccaacaa aggcaatata tgaagatgct aagctctact tatttaacaa tgctacagat 5820

gttagcatta ctgcctcact caagatttgg caaatgaatt ctgcaaatat acaatctagt 5880

taa 5883

<210> 4

<211> 645

<212> PROTEIN

<213> Nicotiana tabacum

<400> 4

Met Ala Glu Thr Asn Asn Ser Val Pro Tyr Thr Gln Leu Pro Ala Glu

1 5 10 15

Asp Asn Asn Thr Ser Ser Asn Ser Pro Ala Lys Cys Arg Arg Arg Pro

20 25 30

Lys Arg Val Ser Phe Ile Val Leu Thr Gly Leu Val Ala Ala Leu Leu

35 40 45

Leu Phe Val Ala Val Lys Tyr Gly Asn Asn Glu Ala Glu Asp Val Asn

50 55 60

Pro Gly Pro Val Pro Pro Gln Glu Thr Val Cys Asn Met Leu Gly Ser

65 70 75 80

Asn Leu Met Pro Leu Thr Thr Met Arg Thr Val Ala Arg Gly Val Ala

85 90 95

Glu Gly Val Ser Ala Lys Ser Arg Gly Arg Phe Leu Gly Leu Arg Pro

100 105 110

Phe Pro Trp Thr Lys Gln Met Leu Ala Trp Gln Arg Thr Ser Phe His

115 120 125

Phe Gln Pro Lys Lys Asn Trp Met Asn Gly Pro Leu Phe Tyr Lys Gly

130 135 140

Trp Tyr His Leu Phe Tyr Gln Tyr Asn Pro Glu Ala Ala Val Trp Gly

145 150 155 160

Asn Ile Val Trp Gly His Ala Val Ser Arg Asp Leu Ile His Trp Gln

165 170 175

His Leu Pro Val Ala Met Val Ala Asp Gln Trp Tyr Asp Ile Asn Gly

180 185 190

Val Trp Thr Gly Ser Ala Thr Ile Leu Pro Asp Gly Lys Leu Val Met

195 200 205

Leu Tyr Thr Gly Ser Thr Asn Glu Ser Val Gln Val Gln Asn Leu Ala

210 215 220

Tyr Pro Ala Asp Pro Ser Asp Pro Leu Leu Arg Lys Trp Val Lys Tyr

225 230 235 240

Glu Gly Asn Pro Val Leu Val Pro Pro Pro Gly Ile Ala Thr Lys Asp

245 250 255

Phe Arg Asp Pro Thr Thr Ala Trp Thr Thr Pro Gln Gly Lys Trp Arg

260 265 270

Ile Thr Ile Gly Ser Lys Val Asn Lys Thr Gly Ile Ser Leu Val Tyr

275 280 285

Asp Thr Ile Asp Phe Lys Lys Phe Glu Leu Leu Asp Gly Val Leu His

290 295 300

Gly Val Pro Gly Thr Gly Met Trp Glu Cys Val Asp Phe Tyr Pro Val

305 310 315 320

Ser Lys Val Val Glu Asn Gly Leu Asp Thr Ser Asp Asn Gly Pro Ala

325 330 335

Val Lys His Val Leu Lys Ser Ser Leu Asp Asp Asp Arg Asn Asp Tyr

340 345 350

Tyr Ala Leu Gly Thr Tyr Asp Ala Val Ala Gly Lys Trp Ile Pro Asp

355 360 365

Asn Pro Thr Ile Asp Val Gly Ile Gly Leu Arg Tyr Asp Tyr Gly Asn

370 375 380

Phe Tyr Ala Ser Lys Thr Phe Tyr Asp Gln Glu Lys Lys Arg Arg Val

385 390 395 400

Leu Trp Ala Trp Ile Thr Glu Gly Asp Ser Glu Ala Ala Asp Ile Cys

405 410 415

Lys Gly Trp Ala Ser Leu Gln Pro Ile Pro Arg Thr Ile Lys Tyr Asp

420 425 430

Lys Lys Thr Gly Ser Asn Ile Ile Thr Trp Pro Val Ala Glu Val Glu

435 440 445

Asn Leu Arg Leu Asn Ser Lys Glu Phe Asp Lys Val Glu Val Lys Pro

450 455 460

Gly Ser Val Phe Pro Leu Glu Val Gly Thr Ala Thr Gln Leu Asp Ile

465 470 475 480

Met Ala Glu Phe Glu Ile Asp Pro Lys Val Leu Glu Arg Leu Glu Gly

485 490 495

Asn Asn Ala Thr Tyr Glu Cys Arg Ser Ser Gly Gly Ser Ala Glu Arg

500 505 510

Gly Ala Leu Gly Pro Phe Gly Leu Leu Val Leu Thr Asp Lys Gly Leu

515 520 525

Ser Glu Gln Thr Pro Ile Tyr Phe Tyr Ile Ala Lys Asp Ala Ala Gly

530 535 540

Asn Phe Thr Thr Phe Phe Cys Asn Asp Leu Thr Arg Ser Ser Glu Ala

545 550 555 560

Thr Asp Val Arg Lys Leu Ile Tyr Gly Ser Thr Val Pro Val Leu Gln

565 570 575

Gly Glu Lys Leu Ser Leu Arg Thr Leu Val Asp His Ser Ile Val Glu

580 585 590

Ser Phe Ala Gln Ser Gly Arg Thr Ala Ile Thr Ser Arg Val Tyr Pro

595 600 605

Thr Lys Ala Ile Tyr Glu Asp Ala Lys Leu Tyr Leu Phe Asn Asn Ala

610 615 620

Thr Asp Val Ser Thr Ala Ser Leu Lys Ile Trp Gln Met Asn Ser Ala

625 630 635 640

Asn Ile Gln Ser Ser

645

<210> 5

<211> 4155

<212> DNA

<213> Nicotiana tabacum

<400> 5

atggccaccc accattccca ttatgacccg gaaaactcca cgacccatta cactgtccta 60

ccggatcaac ccgaatccgc cggcgccggg cgccggaagt ctcttaaagt tgtctccggc 120

attttgctct cctctttctt tttgctttct ttagtctttg tgatcctcaa ccagtcttca 180

gatttatcac aagaaaactc ccgctcgtcg gagactttga cgccggcgtt gtcacgaggt 240

gtatctcagg gagtttccga gaagactttc aaggatgttt ccggtagaag cctttcgtac 300

tacccgtgga ctaatgctat gcttacttgg caaaggactg cttaccattt tcaacctcaa 360

aagaattgga tgaacggtaa attttttggc ttatctttct cttattaatt cttttaataa 420

aacatgaatt ttaagatact tatactggct ttttcttatt gattcttatg gctattttgt 480

tggggtatcc tatggattct gattggatga tatgctgcag atcctaatgg tgagtttact 540

tattaccata attacttttt attatttatt attcccaaac catgattagt gcatccggct 600

attggttaaa gattcacaaa accaataaaa tagtaatctt gtcatagttt ccataataat 660

ctacacgtac gctattgttt aatgacaaga aatttgacgc tcagcatagt taattctctc 720

atatttgtat tgtttacttt atagcctttg agctaattaa ttctgggttt ctttgaacta 780

aacctttata agttacaatc acacatagat gagttggcac attattcagg ctaataatga 840

aagaaattgg attacttgac taatatggca aatgcggcca atttaatttg gattaacacg 900

atatatgtgt ggtaataatg cttttgtgca acatctctca tacaaggaca catgattagg 960

tgattttgta ccaagtctcg ggaccaatca caatatatgg gtcacacctt atatattatt 1020

gtaagagttg ggacccacca aggatttgtc tgtctttcca actagccact tgtctttttc 1080

tcttttttat atttttaaat gaaatggtgt gggtttttta tttttgggtc gatctaaccg 1140

cttctgccta ttatcaattt agccttgtga ttgtgagaat agaagagaga aatagaggat 1200

aataataata aggataagaa ttaagaacgt accttcttat tgtcgaaatt atttgagaag 1260

actattcatt gttctgatta gtgtccatcg atgtcccttt cctccttttt ctatcttgga 1320

gaggtttcct cttctttgtt ttacttttcc tttttctaaa tatgcattcc aaaatcttaa 1380

cactactcga acgtccatc ttggaaagtc tctttgaaag tttagggcaa catcattcgg 1440

acaacttaat tagcattcac tattaaaaat taatagaaca gaaaagttca tgtatttttt 1500

tagggagagt aagaggcgga ttcagaattt aaatcttatg tgtttagttt ttaaaatttt 1560

taggattgat aactgaacat ggcgagaaac atgaacatgt gacataaatt ccgtgtttca 1620

acactaaaca ggtccattat accacaaagg atggtaccat cttttttatc aatacaatcc 1680

tgattcagct gtttggggaa atatcacatg gggccatgca atatccacgg acttgatcca 1740

ctggctttac ttgcctttcg ccatggttcc gatcaatggt acgatatcaa cggtgtctgg 1800

accgggtccg cgaccatctt gcccgacggt cagatcatga tgctatacac cggtgatacc 1860

aatgattacg tgcaggtgca aaatcttgca taccctgcta acttatcgga tcctctcctc 1920

atcgactggg tcaagtacca ggacaatccg gtcatggttc ccccacccgg cattggtgtc 1980

aaggacttca gagacccgac aactgcttgg accggacccc aaaacgggca gtggctgcta 2040

accatcgggt ccaagattgg taaaacgggt attgcacttg tttatgatac gtccaacttc 2100

acaaacttta agctattgga tggagttttg catgcggttc cgggtacggg tatgtgggag 2160

tgtgtggact tttacccggt atcaaccgtt gaggcaaacg ggttggacac atcatataac 2220

gggccaggta taagcatgtg ttaaaagcaa gtttagatga cgataagcat gattactatg 2280

ctattgggac atatgacccg gtaaagaaca aatggactcc tgataacccg gaattggatg 2340

tgggtatcgg gttgagactg gactacggga aatactatgc gtcaaagaca ttttatgacc 2400

cgaaagaaca aagaagaata ttgtggggat ggattggaga aactgacagt gaagctgctg 2460

atctgctgaa gggatgggca tctgtacagg tatggactct tttaagtaca ctacctcagc 2520

atccgaagag cattacactt ttatttttgt tttacattag accacatgaa tgggtgtttt 2580

ggcataactg gtaaagttgt tgccatgtga ccctgaggtc acgggttcga gccgtagaaa 2640

tagcctcttg cagtaatgta ataaactctt agtgcatagg gttgcctttt ttattagacc 2700

acacacatgt tcaagttatg tcatgttagt cgtgtcaatt ttttgtggaa atcaatttac 2760

tgcacctcaa tcttgaatta gttgagacta gctataggaa cctttgtatt gagaggactt 2820

atcataattt gatcattttt gcactaactg tcacactatg atattcactt tctttatcca 2880

gtttagtagt gtgccaatac accttaagca cgtgacaaga atttattagc agggtcatct 2940

cgattttatg taggagtaca gaattgaatt gaatcttttc ttctagtaaa ttctcaattg 3000

caacttgaca atgaagtttt tcagatgcaa aaaagatgaa atatctctaa taatttcctt 3060

ttccaataac agagtattcc aaggactctg ctttatgaca aggagacaag gacacatgta 3120

cttcagtggc cagttaaaga aattgagagc ttaagaattg gtgatcctct agtgaaacag 3180

gtcaatcttc aaccaggctc aattgagctt gtccatgttg actcagccgc acaggtttgc 3240

tttctcatcc ttcgaaattg aaaacgtttc acttatatgt gcttgatgta cagtcctaaa 3300

acttgtatgc gcaatggtgc agttggatgt agaagcctca tttgaagtgg acaaagcagc 3360

actcgcggga acaattgaag cagatgtggt ttcaactgca gtactagtgg aggtgctgct 3420

aaaagaggca ttttgggacc atttggtgtc gttgtaattg ctgatcaaac gctttctgag 3480

ctaaccccag tttacttcta cattgccaaa ggaactggtg gccgagctga aacctacttc 3540

tgcgctgatg aaactaggtt tgcttctact atgtttatct tgtatactct atcttaatag 3600

tccttgtcaa agtatagagg aataacatag cggcgtgatc tgatgcagat cctcagaggc 3660

tcctggagtt gctaaacaag tgtatggtag ttcagtacca gtgttagatg gtgaacaaca 3720

ctcaatgaga ttattggtaa gtgataatcc ctttattctg actttcttca aatcaagaat 3780

aatatcaagc ttattagttc ttccagtcat cttacttaat ttgtggaaat gctccaaagt 3840

agtcaatttg gtaactattc aagataatgt ggttcagaat aatttgtgtt atgaatgtat 3900

ttgacagttg ggatgatctg ttttttagta aaatttctta aaaacttaat tcaggtggac 3960

cactcaattg tggaaagctt tgctcaagga ggaagaacag tcataacatc gcgaatttac 4020

ccaacaaaag caatcaatgg agcagcacga ctgttcgttt tcaacaatgc caccggggct 4080

agtgtgactg cctccctcaa gatttggtca ctcaaatcag ctgatattcg atccttcccc 4140

ttggaccagt tgtaa 4155

<210> 6

<211> 642

<212> PROTEIN

<213> Nicotiana tabacum

<400> 6

Met Ala Thr His His Ser His Tyr Asp Pro Glu Asn Ser Thr Thr His

1 5 10 15

Tyr Thr Val Leu Pro Asp Gln Pro Glu Ser Ala Gly Ala Gly Arg Arg

20 25 30

Lys Ser Leu Lys Val Val Ser Gly Ile Leu Leu Ser Ser Phe Phe Leu

35 40 45

Leu Ser Leu Val Phe Val Ile Leu Asn Gln Ser Ser Asp Leu Ser Gln

50 55 60

Glu Asn Ser Arg Ser Ser Glu Thr Leu Thr Pro Ala Leu Ser Arg Gly

65 70 75 80

Val Ser Gln Gly Val Ser Glu Lys Thr Phe Lys Asp Val Ser Gly Arg

85 90 95

Ser Leu Ser Tyr Tyr Pro Trp Thr Asn Ala Met Leu Thr Trp Gln Arg

100 105 110

Thr Ala Tyr His Phe Gln Pro Gln Lys Asn Trp Met Asn Asp Pro Asn

115 120 125

Gly Pro Leu Tyr His Lys Gly Trp Tyr His Leu Phe Tyr Gln Tyr Asn

130 135 140

Pro Asp Ser Ala Val Trp Gly Asn Ile Thr Trp Gly His Ala Ile Ser

145 150 155 160

Thr Asp Leu Ile His Trp Leu Tyr Leu Pro Phe Ala Met Val Pro Asp

165 170 175

Gln Trp Tyr Asp Ile Asn Gly Val Trp Thr Gly Ser Ala Thr Ile Leu

180 185 190

Pro Asp Gly Gln Ile Met Met Leu Tyr Thr Gly Asp Thr Asn Asp Tyr

195 200 205

Val Gln Val Gln Asn Leu Ala Tyr Pro Ala Asn Leu Ser Asp Pro Leu

210 215 220

Leu Ile Asp Trp Val Lys Tyr Gln Asp Asn Pro Val Met Val Pro Pro

225 230 235 240

Pro Gly Ile Gly Val Lys Asp Phe Arg Asp Pro Thr Thr Ala Trp Thr

245 250 255

Gly Pro Gln Asn Gly Gln Trp Leu Leu Thr Ile Gly Ser Lys Ile Gly

260 265 270

Lys Thr Gly Ile Ala Leu Val Tyr Asp Thr Ser Asn Phe Thr Asn Phe

275 280 285

Lys Leu Leu Asp Gly Val Leu His Ala Val Pro Gly Thr Gly Met Trp

290 295 300

Glu Cys Val Asp Phe Tyr Pro Val Ser Thr Val Glu Ala Asn Gly Leu

305 310 315 320

Asp Thr Ser Tyr Asn Gly Pro Gly Ile Lys His Val Leu Lys Ala Ser

325 330 335

Leu Asp Asp Asp Lys His Asp Tyr Tyr Ala Ile Gly Thr Tyr Asp Pro

340 345 350

Val Lys Asn Lys Trp Thr Pro Asp Asn Pro Glu Leu Asp Val Gly Ile

355 360 365

Gly Leu Arg Leu Asp Tyr Gly Lys Tyr Tyr Ala Ser Lys Thr Phe Tyr

370 375 380

Asp Pro Lys Glu Gln Arg Arg Ile Leu Trp Gly Trp Ile Gly Glu Thr

385 390 395 400

Asp Ser Glu Ala Ala Asp Leu Leu Lys Gly Trp Ala Ser Val Gln Ser

405 410 415

Ile Pro Arg Thr Leu Leu Tyr Asp Lys Glu Thr Arg Thr His Val Leu

420 425 430

Gln Trp Pro Val Lys Glu Ile Glu Ser Leu Arg Ile Gly Asp Pro Leu

435 440 445

Val Lys Gln Val Asn Leu Gln Pro Gly Ser Ile Glu Leu Val His Val

450 455 460

Asp Ser Ala Ala Gln Leu Asp Val Glu Ala Ser Phe Glu Val Asp Lys

465 470 475 480

Ala Ala Leu Ala Gly Thr Ile Glu Ala Asp Val Gly Phe Asn Cys Ser

485 490 495

Thr Ser Gly Gly Ala Ala Lys Arg Gly Ile Leu Gly Pro Phe Gly Val

500 505 510

Val Val Ile Ala Asp Gln Thr Leu Ser Glu Leu Thr Pro Val Tyr Phe

515 520 525

Tyr Ile Ala Lys Gly Thr Gly Gly Arg Ala Glu Thr Tyr Phe Cys Ala

530 535 540

Asp Glu Thr Arg Ser Ser Glu Ala Pro Gly Val Ala Lys Gln Val Tyr

545 550 555 560

Gly Ser Ser Val Pro Val Leu Asp Gly Glu Gln His Ser Met Arg Leu

565 570 575

Leu Val Asp His Ser Ile Val Glu Ser Phe Ala Gln Gly Gly Arg Thr

580 585 590

Val Ile Thr Ser Arg Ile Tyr Pro Thr Lys Ala Ile Asn Gly Ala Ala

595 600 605

Arg Phe Val Phe Asn Asn Ala Thr Gly Ala Ser Val Thr Ala Ser Leu

610 615 620

Lys Ile Trp Ser Leu Lys Ser Ala Asp Ile Arg Ser Phe Pro Leu Asp

625 630 635 640

Gln Leu

<210> 7

<211> 1919

<212> DNA

<213> Nicotiana tabacum

<400> 7

atgatgttat acaccggtga taccaatgat tacgtgcagg tgcaaaatct tgcgtacccc 60

gccaacttat cggatcccct cctcatcgac tgggtcaagt accggggcaa cccggtcatg 120

gttccaccac ccggcattgg tgtcaaggac tttagagacc caacgactgc ttggaccgga 180

ccacaaaacg ggcagtggct gcttaccatc gggtccaaga ttggtaaaac gggtattgca 240

attgtttatg gtacttccaa cttcacaaac tttaagctat tggatggagt tttgcatgcg 300

gttccgggta cgggtatgtg ggagtgtgtg gacttttacc cggtatcaac cgatgaggca 360

aacgggttgg acacatcata taacgggcca ggtataaagc atgtgttaaa agcaagttta 420

gatgacgata agcatgatta ctatgctatt gggacatatg accggtaaag aacaaatgga 480

ctcctgataa cccgcaattg gatgtgggta tcgggttgag actggactac gggaaatact 540

atgcgtcaaa gacattttat gacccgaagg aacaaagaag aatattgtgg ggatggattg 600

gggaaactga cagtgaagct gctgatctgc tgaagggatg ggcatctgta caggtatgga 660

cacttttcaa gtacactacc tcagcttccg aagagcatta cacatttatt tttgtattac 720

attagggtgc cttggcgtaa ctctggtaaa gtaagttctg aattgcaacg tgaaaatgga 780

ggtttttaga tgcaaagaga tgatatatcc ctaatagttt tcctgtttta ataacagagt 840

attccaagga ctgtgcttta tgataaggag actaggacac atgttcttca gtggccagtt 900

aaagaaattg agagcttaag aattggtgat cctctagtga aacgggtcaa tcttcaacca 960

ggctcaattg agctagtcca tgttgactca gccgcacagg ttgctttctc atccttggaa 1020

attgaaaacg tttcacttat atgtgcttaa tgtgcagtcc taaaacttgt atgtgcaatg 1080

gtgcagttgg atgtagaagc ctcatttgaa gtggacaaag cagcactcga gggaacaatt 1140

gaagcagatg ttggtttcaa ctgcagtact agtggaggtg ctgctaaaag aggcattttg 1200

ggaccatttg gtgtcgttgt aattgctgat caaacgcttt ctgagctaac tccagtttac 1260

ttctacattg ccaaaggacc tgatggccga gctgaaacct acttctgtgc tgatgaaact 1320

aggtttgctt ctactatgtt tatcttgtat actctatctt aatagtcctt gtcaaagtat 1380

agatgaataa catagcggcg tgatctgatg cagatcctca gaggctcctg gagttgctaa 1440

acaagtgtat ggtagttcag taccagtgtt agatgatgaa caacactcaa tgagattatt 1500

ggtaagtgat aatcccgtta ttctgacctt cgtcaaatca gaataatatc aagcttatta 1560

gttcttccag tcatcttatt aaatttatgg aaatgctcca aagtagtcaa tttggtaact 1620

attcaagata atgtggttca gaataatttg tgttatgaat gtatttgaca gttgggatga 1680

tctgtgtttt tgagtaaaat ttcttaaaac tgaactcagg tggaccactc aattgtggag 1740

agctttgctc aaggaggaag aacagtcata acatcgcgaa tttacccaac aaaggcaatc 1800

aatggagcag cacgactgtt cgttttcaac aatgccacga gggcaaggtg actgcctccc 1860

tgaagatttg gtcactcgaa tcagctgata ttcgatcctt ccccttggac cagttgtaa 1919

<210> 8

<211> 638

<212> PROTEIN

<213> Nicotiana tabacum

<400> 8

Met Ala Thr His His Ser His Tyr Asp Pro Glu Asn Ser Thr Thr His

1 5 10 15

Tyr Thr Val Leu Pro Asp Gln Pro Glu Ser Ala Gly Ser Gly His Arg

20 25 30

Lys Ser Leu Lys Val Val Ser Gly Ile Leu Leu Ser Ser Phe Phe Leu

35 40 45

Leu Ser Leu Val Phe Val Ile Val Asn Gln Ser Ser Asp Leu Ser Gln

50 55 60

Lys Asn Ser His Ser Ser Glu Thr Leu Thr Pro Ala Leu Ser Arg Gly

65 70 75 80

Val Ser Gln Gly Val Ser Glu Lys Thr Phe Arg Asp Val Ser Gly Gly

85 90 95

Ser Leu Ser Tyr Tyr Pro Trp Thr Asn Ala Met Leu Thr Trp Gln Arg

100 105 110

Thr Ala Tyr His Phe Gln Pro Gln Lys Asn Trp Met Asn Gly Pro Leu

115 120 125

Tyr His Lys Gly Trp Tyr His Leu Phe Tyr Gln Tyr Asn Pro Asp Ser

130 135 140

Ala Ile Trp Gly Asn Ile Thr Trp Gly His Ala Ile Ser Thr Asp Leu

145 150 155 160

Ile His Trp Leu Tyr Leu Pro Phe Ala Leu Val Pro Asp Gln Trp Tyr

165 170 175

Asp Ile Asn Gly Val Trp Thr Gly Ser Ala Thr Phe Leu Pro Asp Gly

180 185 190

Gln Ile Met Met Leu Tyr Thr Gly Asp Thr Asn Asp Tyr Val Gln Val

195 200 205

Gln Asn Leu Ala Tyr Pro Ala Asn Leu Ser Asp Pro Leu Leu Ile Asp

210 215 220

Trp Val Lys Tyr Arg Gly Asn Pro Val Met Val Pro Pro Pro Gly Ile

225 230 235 240

Gly Val Lys Asp Phe Arg Asp Pro Thr Thr Ala Trp Thr Gly Pro Gln

245 250 255

Asn Gly Gln Trp Leu Leu Thr Ile Gly Ser Lys Ile Gly Lys Thr Gly

260 265 270

Ile Ala Ile Val Tyr Gly Thr Ser Asn Phe Thr Asn Phe Lys Leu Leu

275 280 285

Asp Gly Val Leu His Ala Val Pro Gly Thr Gly Met Trp Glu Cys Val

290 295 300

Asp Phe Tyr Pro Val Ser Thr Asp Glu Ala Asn Gly Leu Asp Thr Ser

305 310 315 320

Tyr Asn Gly Pro Gly Ile Lys His Val Leu Lys Ala Ser Leu Asp Asp

325 330 335

Asp Lys His Asp Tyr Tyr Ala Ile Gly Thr Tyr Asp Pro Val Lys Asn

340 345 350

Lys Trp Thr Pro Asp Asn Pro Gln Leu Asp Val Gly Ile Gly Leu Arg

355 360 365

Leu Asp Tyr Gly Lys Tyr Tyr Ala Ser Lys Thr Phe Tyr Asp Pro Lys

370 375 380

Glu Gln Arg Arg Ile Leu Trp Gly Trp Ile Gly Glu Thr Asp Ser Glu

385 390 395 400

Ala Ala Asp Leu Leu Lys Gly Trp Ala Ser Val Gln Ser Ile Pro Arg

405 410 415

Thr Val Leu Tyr Asp Lys Glu Thr Arg Thr His Val Leu Gln Trp Pro

420 425 430

Val Lys Glu Ile Glu Ser Leu Arg Ile Gly Asp Pro Leu Val Lys Arg

435 440 445

Val Asn Leu Gln Pro Gly Ser Ile Glu Leu Val His Val Asp Ser Ala

450 455 460

Ala Gln Leu Asp Val Glu Ala Ser Phe Glu Val Asp Lys Ala Ala Leu

465 470 475 480

Glu Gly Thr Ile Glu Ala Asp Val Gly Phe Asn Cys Ser Thr Ser Gly

485 490 495

Gly Ala Ala Lys Arg Gly Ile Leu Gly Pro Phe Gly Val Val Val Ile

500 505 510

Ala Asp Gln Thr Leu Ser Glu Leu Thr Pro Val Tyr Phe Tyr Ile Ala

515 520 525

Lys Gly Pro Asp Gly Arg Ala Glu Thr Tyr Phe Cys Ala Asp Glu Thr

530 535 540

Arg Ser Ser Glu Ala Pro Gly Val Ala Lys Gln Val Tyr Gly Ser Ser

545 550 555 560

Val Pro Val Leu Asp Asp Glu Gln His Ser Met Arg Leu Leu Val Asp

565 570 575

His Ser Ile Val Glu Ser Phe Ala Gln Gly Gly Arg Thr Val Ile Thr

580 585 590

Ser Arg Ile Tyr Pro Thr Lys Ala Ile Asn Gly Ala Ala Arg Leu Phe

595 600 605

Val Phe Asn Asn Ala Thr Arg Ser Val Thr Ala Ser Leu Lys Ile Trp

610 615 620

Ser Leu Glu Ser Ala Asp Ile Arg Ser Phe Pro Leu Asp Gln

625 630 635

<210> 9

<211> 98

<212> DNA

<213> Artificial sequence

<220>

<223> Nucleotide sequence for silencing of NtINV4-T and NtINV4-S

<400> 9

ggtttcaact gcagtactag tggaggtgct gctaaaagag gcattttggg accatttggt 60

gtcgttgtaa ttgctgatca aacgctttct gagctaac 98

<210> 10

<211> 4384

<212> DNA

<213> Nicotiana tabacum

<220>

<221> general category

<222> (1520)..(1520)

<223> n is a, c, g, or t

<400> 10

atggcagcta gtggtcttag cattaagaaa agtttggagg aatccattt ggctcatcca 60

gatgaaattt tggctctcaa gtcaaggtac attactacat ataatgatat taagaactag 120

aggcttatcc aaggttttgt tacatttttg aaattataag tttagaacct aatagtactt 180

ggtagcactt gtttccttat tatctagctg ttgttactgc ttgttgctac tgctttctgt 240

tcatctttcc ttgagcccgg tctatcggaa acaacctctc tattctcaaa gtataaggtt 300

tgcgtacata ctacctcccc agactctact tgtggaattt actgtttttg ttgtgttgtt 360

gtaatctaat atttattaga attttactga tttttcacat atatatatct atgtcccctg 420

tcgaaaattc tatagctcat gttagctaaa tacattagta ccattgtttt taattgtttt 480

ggttttggca caggattgaa actgaaggga aaggggtaat gaaaccactt gatctcttga 540

accatttggt ttctgttact agtaagacaa atggagtaaa tattgtacct agtgcacttg 600

tggaagttct cagttgcagc caagaagctg tgattgtacc accaaaacta gcactagctg 660

tacgtccgag gcccggtgta tgggagtact tgtcactgaa tcttaagaca aagaaagtgg 720

ctgaattaag cattcctgaa taccttcaat tgaaagagaa cactgttgat gaaaggtaaa 780

gtattagtct gcgatttcgc tttgtgaaat tgaagttttt gttttgattc ataatgtttt 840

gtgtatcaat tatgttacca gtggaaacat attggagttg gattttgagc catttacaac 900

agttacacca ccaaaaacac tttctgactc tattggcaat ggtttggagt ttcttaatcg 960

ccacattgct tcgaaaatgt ttcatgataa ggagatttcc agatgcctcc ttgacttcct 1020

cagaaaccat aactacaaag gaaaggtaat aaaaaaaagt gtttctttaa acaagttgta 1080

tgattatgtg tatatttcta agtatgttaa cttgaaaaca gtcattgatg gtgaaagaaa 1140

gcattcaaag cctagagagt ttccaacttg ttctgaaaaa agcagaggaa catttgtgca 1200

cattgaatcc agaaactcca tactccaatt ttgaatcaaa gtttgaagag attggcttgg 1260

aaagaggtg gggaaacacc gctgaacgcg tgcaagacac tatcagtcat cttttgcatc 1320

tccttgaggc tcctaacgcg tcttctttgg aaaatttcct tggtagaatc ccattggttt 1380

tcaatgttgt gattctaact ccacatggtt attttgctca agataatgtc ttgggctatc 1440

ctgacactgg tggccaggtt tgtgtccaat attttgcatt cttgatcaag ttctttatac 1500

catttgaacc aacaatcttn aacattcttt ttttggttgt gaaatgttga ataggttgtt 1560

tacattcttg atcaagttcc agctatggag cgtgagatgc ttcatcgtat gaagcttcaa 1620

ggactcgatg atatcatccc tcgcatcctt gttgtaagtg gccttaattt tcctagtttc 1680

atttacacct ctaaatgaaa ttgatctttt ttgttgtttt atatcaggta acaaggctgc 1740

tgcctgatgc agtaggaacc acctgtggcg agcggatgga gaaagtatat ggggcagaac 1800

attctcatat aattcgtgtt ccatttagaa ctgagaaggg aatgttgcgc aaatggatct 1860

cacgattcga agtctggcca tacatggaaa ctttcactga ggttggaaca taaaaacaaa 1920

taaaatccat tggaatgttc cttctgcaat tgaaaatgtc ttgctaactg aagacccatt 1980

tttaaattga tcatcaggat gttgcagaag aacttgtcaa agaattgcaa gctaaaccag 2040

acttgatcat tggaaactac agtgagggaa atcttgctgc ctctttgctt gcgaagaaat 2100

ttggggctac tcagtgtact attgctcatg ccttggaaaa aactaagtat ccaaactctg 2160

accttaattg gaagaagttt gatgacaagt atcatttctc aagtcagttc actgctgatc 2220

tctttgccat gaatcacact gatttcatca tcaccagcac tttccaagaa attgctggaa 2280

ggtaaaagca aatgcacacc atcatagtat ttcatatttt tacccttgtt tatactattt 2340

ccattcaccg accccgactt gtttaggatt gagccatagt tgttgttgtt gtttgtttat 2400

actatttcca tttgccgacc acaacttgtt taggactgag gtatagttgt tgttgttggt 2460

ttgttcatat tattttcatt cgctaaccct aacttgtttg ggactgaggc atagtagtag 2520

tagtagttgt tgctattagt ttatactatt tccatttgcc aaccccaact tgtttggtac 2580

tgagacatag ttgttgttgt tgttgtttgt ttatactatt tccatttgcc gaccccaact 2640

tgtttaggac tgaggtatag ttgttgttgt tggtttgttc atattatttt cattcgctaa 2700

ccccaacttg tttgggactg aggcatagta gtagtagtag tagttgttgc tattagttta 2760

tactatttcc atttgccaac cccaacttgt ttggtactga gacatagttg ttgttgttgt 2820

ttgtttatac tatttcaatt tgtcgacccc aatttgtttg ggaccaaggc atggttgttg 2880

ttgttgtttg tttgttttta ctgtttccat tgatattgga acatttgtta tttgcagcaa 2940

aaacactgta ggacagtatg agagtcatac tgcttttacc atgcctggat tgtaccgagt 3000

agtccatgga atcgattcgt ttgatccaaa gttcaacatt gtctcccctg gggctgatat 3060

gtcaatctac ttcccttaca ctgagaagga gaaaaggcta accaacttcc acccggaaat 3120

tgaagaactc ctctacagtc ctgttgagaa taaggaccac ttgttagtct ccttaatttg 3180

cttttatttc atcccattta tgatcgcttt tatcccaaca gatcgattaa tcatttgtta 3240

tcaacataaa cagatgtgtg ttgaaggacc ggaacaagcc aattctcttt accatggcaa 3300

ggctagatcg cgtgaagaat ctaacagggc tcgtggaatg gtatgctaag aatgcaaggc 3360

tgagggagct tgttaacctt gtggttgtag gcggagacag aaggaaagaa tccaaagatt 3420

tagaagagca agcagagatg aagaagatgt atgatcttat cgaaacctat aacctgaacg 3480

gccaattcag gtggatttct tcccaaatga atcgtgtgag gaacggagaa ctctatcgtt 3540

acattgcaga cacgaggggt gctttcgttc aaccagcatt ctacgaggct tttggtttga 3600

cagttgtaga gtctatgact tgtggtttgc caacttttgc tacttgtaat ggtggaccat 3660

ttgagattat agtgaatgga aaatctggtt tccatattga tcctaatcaa ggtgacaagg 3720

ctgctgatat gttggtaaat ttctttgaaa aatctaaaga agatccaagt tattgggatg 3780

ctatttccaa gggaggtctg caacgtattc ttgaaaagta agcttttgca tttgattagc 3840

acaagtgcac aaccaagatt taacttttga acaaactaaa actaaccctt ttttgtattt 3900

tcttttgcta ggtatacatg gcaaatttat tcacagaaag tgatcacact atctgggatt 3960

tatggattct ggaagtatgc aaccaagaat gataaagttg ctagtgcaaa gaagcgctat 4020

cttgagatgt tttatgaact tggatttaag aaatcagtaa gtgtcaattt taaaggggaa 4080

ccttggatca acggttaagt tgtctttgtg caacctatag gtcaggggtt tgagccgtag 4140

aagtagccac taatatttac attagggtag actgtgtaca tatcacaccc cttggggtac 4200

ggccctttcc tggatcctgt atgaacgcgg gatgccttgt gcaccgggct gtattttttt 4260

ttttagtgtc acttctgtat tttgtttgag cttgtttata aagtttggaa atctgctgct 4320

aatttgtata tttgttggtt gtgtatttca ggctgagaaa gttccattgg ctattgatga 4380

atag 4384

<210> 11

<211> 803

<212> PROTEIN

<213> Nicotiana tabacum

<400> 11

Met Ala Ala Ser Gly Leu Ser Ile Lys Lys Ser Leu Glu Glu Ser Ile

1 5 10 15

Leu Ala His Pro Asp Glu Ile Leu Ala Leu Lys Ser Arg Ile Glu Thr

20 25 30

Glu Gly Lys Gly Val Met Lys Pro Leu Asp Leu Leu Asn His Leu Val

35 40 45

Ser Val Thr Ser Lys Thr Asn Gly Val Asn Ile Val Pro Ser Ala Leu

50 55 60

Val Glu Val Leu Ser Cys Ser Gln Glu Ala Val Ile Val Pro Pro Lys

65 70 75 80

Leu Ala Leu Ala Val Arg Pro Arg Pro Gly Val Trp Glu Tyr Leu Ser

85 90 95

Leu Asn Leu Lys Thr Lys Lys Val Ala Glu Leu Ser Ile Pro Glu Tyr

100 105 110

Leu Gln Leu Lys Glu Asn Thr Val Asp Glu Ser Gly Asn Ile Leu Glu

115 120 125

Leu Asp Phe Glu Pro Phe Thr Thr Val Thr Pro Pro Lys Thr Leu Ser

130 135 140

Asp Ser Ile Gly Asn Gly Leu Glu Phe Leu Asn Arg His Ile Ala Ser

145 150 155 160

Lys Met Phe His Asp Lys Glu Ile Ser Arg Cys Leu Leu Asp Phe Leu

165 170 175

Arg Asn His Asn Tyr Lys Gly Lys Ser Leu Met Val Lys Glu Ser Ile

180 185 190

Gln Ser Leu Glu Ser Phe Gln Leu Val Leu Lys Lys Ala Glu Glu His

195 200 205

Leu Cys Thr Leu Asn Pro Glu Thr Pro Tyr Ser Asn Phe Glu Ser Lys

210 215 220

Phe Glu Glu Ile Gly Leu Glu Arg Gly Trp Gly Asn Thr Ala Glu Arg

225 230 235 240

Val Gln Asp Thr Ile Ser His Leu Leu His Leu Leu Glu Ala Pro Asn

245 250 255

Ala Ser Ser Leu Glu Asn Phe Leu Gly Arg Ile Pro Leu Val Phe Asn

260 265 270

Val Val Ile Leu Thr Pro His Gly Tyr Phe Ala Gln Asp Asn Val Leu

275 280 285

Gly Tyr Pro Asp Thr Gly Gly Gln Val Val Tyr Ile Leu Asp Gln Val

290 295 300

Pro Ala Met Glu Arg Glu Met Leu His Arg Met Lys Leu Gln Gly Leu

305 310 315 320

Asp Asp Ile Ile Pro Arg Ile Leu Val Val Thr Arg Leu Leu Pro Asp

325 330 335

Ala Val Gly Thr Thr Cys Gly Glu Arg Met Glu Lys Val Tyr Gly Ala

340 345 350

Glu His Ser His Ile Ile Arg Val Pro Phe Arg Thr Glu Lys Gly Met

355 360 365

Leu Arg Lys Trp Ile Ser Arg Phe Glu Val Trp Pro Tyr Met Glu Thr

370 375 380

Phe Thr Glu Asp Val Ala Glu Glu Leu Val Lys Glu Leu Gln Ala Lys

385 390 395 400

Pro Asp Leu Ile Ile Gly Asn Tyr Ser Glu Gly Asn Leu Ala Ala Ser

405 410 415

Leu Leu Ala Lys Lys Phe Gly Ala Thr Gln Cys Thr Ile Ala His Ala

420 425 430

Leu Glu Lys Thr Lys Tyr Pro Asn Ser Asp Leu Asn Trp Lys Lys Phe

435 440 445

Asp Asp Lys Tyr His Phe Ser Ser Gln Phe Thr Ala Asp Leu Phe Ala

450 455 460

Met Asn His Thr Asp Phe Ile Ile Thr Ser Thr Phe Gln Glu Ile Ala

465 470 475 480

Gly Ser Lys Asn Thr Val Gly Gln Tyr Glu Ser His Thr Ala Phe Thr

485 490 495

Met Pro Gly Leu Tyr Arg Val Val His Gly Ile Asp Ser Phe Asp Pro

500 505 510

Lys Phe Asn Ile Val Ser Pro Gly Ala Asp Met Ser Ile Tyr Phe Pro

515 520 525

Tyr Thr Glu Lys Glu Lys Arg Leu Thr Asn Phe His Pro Glu Ile Glu

530 535 540

Glu Leu Leu Tyr Ser Pro Val Glu Asn Lys Asp His Leu Cys Val Leu

545 550 555 560

Lys Asp Arg Asn Lys Pro Ile Leu Phe Thr Met Ala Arg Leu Asp Arg

565 570 575

Val Lys Asn Leu Thr Gly Leu Val Glu Trp Tyr Ala Lys Asn Ala Arg

580 585 590

Leu Arg Glu Leu Val Asn Leu Val Val Val Gly Gly Asp Arg Arg Lys

595 600 605

Glu Ser Lys Asp Leu Glu Glu Gln Ala Glu Met Lys Lys Met Tyr Asp

610 615 620

Leu Ile Glu Thr Tyr Asn Leu Asn Gly Gln Phe Arg Trp Ile Ser Ser

625 630 635 640

Gln Met Asn Arg Val Arg Asn Gly Glu Leu Tyr Arg Tyr Ile Ala Asp

645 650 655

Thr Arg Gly Ala Phe Val Gln Pro Ala Phe Tyr Glu Ala Phe Gly Leu

660 665 670

Thr Val Val Glu Ser Met Thr Cys Gly Leu Pro Thr Phe Ala Thr Cys

675 680 685

Asn Gly Gly Pro Phe Glu Ile Ile Val Asn Gly Lys Ser Gly Phe His

690 695 700

Ile Asp Pro Asn Gln Gly Asp Lys Ala Ala Asp Met Leu Val Asn Phe

705 710 715 720

Phe Glu Lys Ser Lys Glu Asp Pro Ser Tyr Trp Asp Ala Ile Ser Lys

725 730 735

Gly Gly Leu Gln Arg Ile Leu Glu Lys Tyr Thr Trp Gln Ile Tyr Ser

740 745 750

Gln Lys Val Ile Thr Leu Ser Gly Ile Tyr Gly Phe Trp Lys Tyr Ala

755 760 765

Thr Lys Asn Asp Lys Val Ala Ser Ala Lys Lys Arg Tyr Leu Glu Met

770 775 780

Phe Tyr Glu Leu Gly Phe Lys Lys Ser Ala Glu Lys Val Pro Leu Ala

785 790 795 800

Ile Asp Glu

<210> 12

<211> 3926

<212> DNA

<213> Nicotiana tabacum

<400> 12

atggcaggca gtggtcttag cattaaggaa agtttggagg aatccattt ggctcatcca 60

gatgaaattt tggctctcaa gtcaaggtac attactgcat aatgatatta agacctagaa 120

gcggatccaa gattttgtta catttttgaa attataagtt tagaatctaa tatttgttat 180

cgcttgtttc cttattatct tgctgttgtt actgcctgtt gctactagtt tctgttcatc 240

cttccttgag ctgagtttct atcggaaaca acctctctac tctcaaagta ggaataagtt 300

atgcgtacac actaccctcc ccagactcca cttgtgtaat ttactgagtt tgttgttgtt 360

gttgttgtaa tctaatactt gttagaattt tactgatttt tcacatatat atctatgacc 420

catgtcgaaa atactatagc tcatgtgcta aatacattag taccattgtt ttgtaattgt 480

tttggttttg gaacaggatt gaaactgaag ggaaaggggt aatgaaacca gttgatctct 540

tgaaccattt ggtttctgtt actagtaaaa caaatggagt aaatgttgta cctagtgcac 600

ttgtggaagt tctcagttgc agccaagaag ctgtgattgt accaccaaaa ctagcactag 660

ctgtacgtcc gaggcccggt gtatgggagt acttgtcact gaatcttaag acaaagaaag 720

tggctgaatt gagcattcct gagtaccttc aattgaaaga gaatactgtt gatgaaaggt 780

aaagtaatag tctgcgattt cgctttgtga aattgaagtt ttttgtttga ttcttaatgt 840

tttgtgtatc aattatgtta ccagtggaaa catcttggag ttggattttg agccatttac 900

aactgttaca acaccaaaaa cactttctga ctctattggc aatggtttgg agtttcttaa 960

tcgccacatt gcttcgaaaa tgtttcttga taaggagatt gccaagtgcc tccttgactt 1020

tctcagaaac cataactaca aaggaaaggt agtaaaaaaa gtgtttcttt aaacaagttg 1080

tatgattatg tgtgtatttc taaatatgtc aatttgaaaa cagtcattga tggtgaaaga 1140

aagcattcaa agcctggaga gtttccaact tgttctgaaa aaagcagagg aatatttgca 1200

cacactgaat ccagaaactc catactccaa atttgaatcc aagtttgaag agattggctt 1260

ggaaagaggg tggggaaaca ccgctgaacg cgtgcaagac accattagtc atcttttgca 1320

tctccttgag gctcctaacg cgtcttcctt ggaaaatttc cttggtagaa tcccattggt 1380

tttcaatgtt gtgattctca ccccacatgg ttattttgct caagataatg tcttgggcta 1440

tcctgacact ggtggccagg tttgtgtccg atataacata tcaagaaatt ttgcattctt 1500

gatcatgttc tttataccat ttgaaccaac attctttttt tggttgtgaa atgttgaata 1560

ggttgtttac attcttgatc aagttccagc tatggagcgt gagatgcttc atcgtatgaa 1620

gcttcaagga ctcgacgata tcatccctcg catccttgtt gtaagtgccc ttaattttcc 1680

tggtttggtt tacctctaaa tgaaattgat tttctggctt tctaactttt ttggattgat 1740

ctttttgttg ttttatatca ggtaactagg ctgctgcctg atgctgtagg aaccacttgt 1800

ggcgagtgga tggagaaagt atatggggca gaacattctc atataattcg tgttccatt 1860

agaactgaga aaggaatgtt gcgcaaatgg atctcacgat tcgaagtctg gccatacatg 1920

gaaactttca ctgaggttgg aacataaaaa caaataaaaa tcattggaat gttcttctgc 1980

atttgaaaat gtcttgctaa ctaaagactc atttttaaat taatcatcag gatgttgcag 2040

aagaacttgt caaagaattg caagctaaac cagacttgat aattggaaac tacagtgagg 2100

gaaatcttgc tgcctcattg cttgctaaga aatttggggc tactcagtgt actattgctc 2160

atgccttgga aaaaactaag tatccaaact ctgaccttaa ttggaagaag tttgatgaca 2220

agtatcattt ctcaagtcag ttcactgctg atctttttgc catgaatcac actgatttca 2280

ttatcaccag cactttccaa gaaattgctg gaaggtaaaa gcaaatgcac accatcatag 2340

tatttcatat ttttacccta gtttatacta tttccatttg tcaactccaa cttgtttggg 2400

attgaaccat agttgttgtt tgtttatact atttccattc gccgacccca acttatttgg 2460

gactgagaca taattgttgt tattattgtt tgtttgttta tactatttcc attctcagac 2520

cccaacttct ttgggactga gccgtagatt gttgttgttg ttgttgttgt tgtttgttta 2580

tgctatttcc gttcaccgac cccaacttat ttgggactga ggtgtagaag tagtcgttgt 2640

tgtttgttta tacgacttcc aattgatatt cgaatgtttt tatttttgca gcaagaacac 2700

tgtaggacag tatgagagtc atactgcttt taccatgcct ggattgtatc gagtagtcca 2760

tggaatcaat tcgtttgatc caaagttcaa cattgtctcc cctggggctg atatgtcaat 2820

ctacttccct tacactgaga aggagaaaag actaaccaac ttccacccgg aaattgaaga 2880

actcctctac agtcctgttg agaataagga ccacttgtta gtcttcttta tttcattcat 2940

ttttctacac cttttttttc aacagattga ttgattggtt cttatcaacg taaacagatg 3000

tgtgttgaag gaccagaaca agccaattct ctttaccatg gcaaggctag atcgcgtgaa 3060

gaatctaaca gggctcgtgg aatggtatgc aaagaatgca aggctaaggg agctcgttaa 3120

ccttgtggtt gtaggcggag acagaaggaa agaatccaaa gatttagaag agcaagcaga 3180

gatgaagaag atgtatgatc ttatcgaaac atacaacctg aatggccaat tcaggtggat 3240

ttcttcccaa atgaatcgtg tgaggaacgg agaactttat cgatacattg cagacacgag 3300

gggtgctttc gttcaaccag cattttatga ggcatttggt ttgacagttg ttgagtctat 3360

gacttgtggt ttgccaactt ttgctacttg taatggtgga ccatttgaga ttatagtgaa 3420

tggaaaatct ggtttccata ttgatcctaa tcaaggtgac aaggctgctg atatgttggt 3480

taatttcttc gaaaaatcta aagaagatcc aagttattgg gatactattt ccaagggtgg 3540

tctgcagcgt attcttgaaa agtaagcttt tgcatttgat tagcacaagt gtacaaccaa 3600

gatttaactt atgaacaaac taaaactaac ccttttttta ttttcttttg ctaggtatac 3660

atggcaaatt tattcacaga aagtgatcac attatctggg atttatggat tctggaaata 3720

tgcaaccaag aatgacaaag ttgctagtgc gaagaagcgc tatcttgaaa tgttttatga 3780

atttgggttt aagaaatcag taagtgtcac ttctgtattt tgtttgagct tgtttgtaaa 3840

gtttggcaat cttctgctaa tttgtactat atttgttgac ttgtgcattt caggctgaga 3900

aagttccatt ggctattgat gaatag 3926

<210> 13

<211> 803

<212> PROTEIN

<213> Nicotiana tabacum

<400> 13

Met Ala Gly Ser Gly Leu Ser Ile Lys Glu Ser Leu Glu Glu Ser Ile

1 5 10 15

Leu Ala His Pro Asp Glu Ile Leu Ala Leu Lys Ser Arg Ile Glu Thr

20 25 30

Glu Gly Lys Gly Val Met Lys Pro Val Asp Leu Leu Asn His Leu Val

35 40 45

Ser Val Thr Ser Lys Thr Asn Gly Val Asn Val Val Pro Ser Ala Leu

50 55 60

Val Glu Val Leu Ser Cys Ser Gln Glu Ala Val Ile Val Pro Pro Lys

65 70 75 80

Leu Ala Leu Ala Val Arg Pro Arg Pro Gly Val Trp Glu Tyr Leu Ser

85 90 95

Leu Asn Leu Lys Thr Lys Lys Val Ala Glu Leu Ser Ile Pro Glu Tyr

100 105 110

Leu Gln Leu Lys Glu Asn Thr Val Asp Glu Ser Gly Asn Ile Leu Glu

115 120 125

Leu Asp Phe Glu Pro Phe Thr Thr Val Thr Thr Pro Lys Thr Leu Ser

130 135 140

Asp Ser Ile Gly Asn Gly Leu Glu Phe Leu Asn Arg His Ile Ala Ser

145 150 155 160

Lys Met Phe Leu Asp Lys Glu Ile Ala Lys Cys Leu Leu Asp Phe Leu

165 170 175

Arg Asn His Asn Tyr Lys Gly Lys Ser Leu Met Val Lys Glu Ser Ile

180 185 190

Gln Ser Leu Glu Ser Phe Gln Leu Val Leu Lys Lys Ala Glu Glu Tyr

195 200 205

Leu His Thr Leu Asn Pro Glu Thr Pro Tyr Ser Lys Phe Glu Ser Lys

210 215 220

Phe Glu Glu Ile Gly Leu Glu Arg Gly Trp Gly Asn Thr Ala Glu Arg

225 230 235 240

Val Gln Asp Thr Ile Ser His Leu Leu His Leu Leu Glu Ala Pro Asn

245 250 255

Ala Ser Ser Leu Glu Asn Phe Leu Gly Arg Ile Pro Leu Val Phe Asn

260 265 270

Val Val Ile Leu Thr Pro His Gly Tyr Phe Ala Gln Asp Asn Val Leu

275 280 285

Gly Tyr Pro Asp Thr Gly Gly Gln Val Val Tyr Ile Leu Asp Gln Val

290 295 300

Pro Ala Met Glu Arg Glu Met Leu His Arg Met Lys Leu Gln Gly Leu

305 310 315 320

Asp Asp Ile Ile Pro Arg Ile Leu Val Val Thr Arg Leu Leu Pro Asp

325 330 335

Ala Val Gly Thr Thr Cys Gly Glu Trp Met Glu Lys Val Tyr Gly Ala

340 345 350

Glu His Ser His Ile Ile Arg Val Pro Phe Arg Thr Glu Lys Gly Met

355 360 365

Leu Arg Lys Trp Ile Ser Arg Phe Glu Val Trp Pro Tyr Met Glu Thr

370 375 380

Phe Thr Glu Asp Val Ala Glu Glu Leu Val Lys Glu Leu Gln Ala Lys

385 390 395 400

Pro Asp Leu Ile Ile Gly Asn Tyr Ser Glu Gly Asn Leu Ala Ala Ser

405 410 415

Leu Leu Ala Lys Lys Phe Gly Ala Thr Gln Cys Thr Ile Ala His Ala

420 425 430

Leu Glu Lys Thr Lys Tyr Pro Asn Ser Asp Leu Asn Trp Lys Lys Phe

435 440 445

Asp Asp Lys Tyr His Phe Ser Ser Gln Phe Thr Ala Asp Leu Phe Ala

450 455 460

Met Asn His Thr Asp Phe Ile Ile Thr Ser Thr Phe Gln Glu Ile Ala

465 470 475 480

Gly Ser Lys Asn Thr Val Gly Gln Tyr Glu Ser His Thr Ala Phe Thr

485 490 495

Met Pro Gly Leu Tyr Arg Val Val His Gly Ile Asn Ser Phe Asp Pro

500 505 510

Lys Phe Asn Ile Val Ser Pro Gly Ala Asp Met Ser Ile Tyr Phe Pro

515 520 525

Tyr Thr Glu Lys Glu Lys Arg Leu Thr Asn Phe His Pro Glu Ile Glu

530 535 540

Glu Leu Leu Tyr Ser Pro Val Glu Asn Lys Asp His Leu Cys Val Leu

545 550 555 560

Lys Asp Gln Asn Lys Pro Ile Leu Phe Thr Met Ala Arg Leu Asp Arg

565 570 575

Val Lys Asn Leu Thr Gly Leu Val Glu Trp Tyr Ala Lys Asn Ala Arg

580 585 590

Leu Arg Glu Leu Val Asn Leu Val Val Val Gly Gly Asp Arg Arg Lys

595 600 605

Glu Ser Lys Asp Leu Glu Glu Gln Ala Glu Met Lys Lys Met Tyr Asp

610 615 620

Leu Ile Glu Thr Tyr Asn Leu Asn Gly Gln Phe Arg Trp Ile Ser Ser

625 630 635 640

Gln Met Asn Arg Val Arg Asn Gly Glu Leu Tyr Arg Tyr Ile Ala Asp

645 650 655

Thr Arg Gly Ala Phe Val Gln Pro Ala Phe Tyr Glu Ala Phe Gly Leu

660 665 670

Thr Val Val Glu Ser Met Thr Cys Gly Leu Pro Thr Phe Ala Thr Cys

675 680 685

Asn Gly Gly Pro Phe Glu Ile Ile Val Asn Gly Lys Ser Gly Phe His

690 695 700

Ile Asp Pro Asn Gln Gly Asp Lys Ala Ala Asp Met Leu Val Asn Phe

705 710 715 720

Phe Glu Lys Ser Lys Glu Asp Pro Ser Tyr Trp Asp Thr Ile Ser Lys

725 730 735

Gly Gly Leu Gln Arg Ile Leu Glu Lys Tyr Thr Trp Gln Ile Tyr Ser

740 745 750

Gln Lys Val Ile Thr Leu Ser Gly Ile Tyr Gly Phe Trp Lys Tyr Ala

755 760 765

Thr Lys Asn Asp Lys Val Ala Ser Ala Lys Lys Arg Tyr Leu Glu Met

770 775 780

Phe Tyr Glu Phe Gly Phe Lys Lys Ser Ala Glu Lys Val Pro Leu Ala

785 790 795 800

Ile Asp Glu

<210> 14

<211> 4532

<212> DNA

<213> Nicotiana tabacum

<400> 14

atggctgaac gtgctctgac tcgtgttcac agccttcgtg aacgtcttga tgccactttg 60

gctgcacatc gcaatgagat attgctgttt ctttcaaggt attgcctaag tagtgttctt 120

gtttcctaca aaagattcag ttggtgttca aaaaacgata tgtgatttga tttatctgcc 180

taagtcttgg tagtcataat tatccggtac ctgtgctggt gcgagttagc tggttcggaa 240

actactctta tgaaaacgag agatttagtt ggtgttgtct gcaattctgt agtatggact 300

attaagcaga tagatcatgt ttgatatcga aaaggaatgt atatgtgatg ttacttgaac 360

tggttttggt tattacagga ttgaaagcca tggaaaaggg atcttgaaac ctcaccagct 420

attggctgag ttcgatgcaa ttcgccaaga tgacaaaaag aagctgaatg atcatgcatt 480

tgaagaactc ctgaaatcta ctcaggtaat tttgattttg gctaaatgtg ttaccaagct 540

gaatgatcat gcatttgagt ttgtgtccga ctactacaat gatatgttat accaggaagc 600

gattgttctg ccaccttggg ttgcacttgc cattcgtttg aggcctggtg tgtgggaata 660

tgtccgtgtg aatgttaatg ctctagtcgt tgaggagctg accgtccctg agtatttgca 720

ttttaaggaa gaacttgttg atggaacgta agttttagtc tcttatttga tactatgtta 780

gagaataggc agtggattca atttatcagt gttgtttttt acctaatgca gctccaatgg 840

aaatttcgtt ctcgagttgg attttgagcc cttcactgca tcctttccta aaccgaccct 900

caccaaatct attgggaatg gagttgaatt cctcaatagg cacctttctg cgaaaatgtt 960

ccatgacaag gaaagcatga ccccgcttct tgaatttctt cgggttcaca attataaggg 1020

caaggtaact ttgttattcc cattcatata tatgttcagt ttgtgcttat catgcgccca 1080

atgatgtatg aatatgtact aaaggataga tgtacgattt cgtttgcaga caatgatgct 1140

gaatgacaga atacagaatt taaccactct gcaaaatgtc ctaaggaagg cagaggaata 1200

ccttattatg cttccccctg aaactccatt ttccgaattc gaacacaagt tccaagaaat 1260

tggattggag aagggatggg gcgacactgc ggagcgcgtg ctagagatga tatgcatgct 1320

tcttgatcta cttgaggctc ccgactcctg tactcttgag aagttcctag ggagaattcc 1380

tatggtgttc aacgtggtta tcctttcccc ccatggatat ttcgcccagg aaaatgtctt 1440

gggttatccc gacactggtg gccaggtgca ttactttagt ctttgtccgt gagtctatgt 1500

tgctcagatc ctctacaatg ccactgtacc cgtgtaggat actccaaata taatgcattt 1560

ttggaggatc tgtcaccggt gcaatggcat tttggaggtc ggagcaacaa acaactgcta 1620

gtatgcttct aaagcttgct tccataaatg ctaaggtcct tcacccgtaa tgtgcaggtt 1680

gtctacatat tagatcaagt tccagccttg gagcgtgaaa tgcttaaacg cctaaaggag 1740

caaggacttg atataacacc gcgtattctt attgttagta tttcttgtac ttgtaattgc 1800

tgcggattac acaaaatttt ctctttattg gcaacttatc ttgatattat tcccaggtta 1860

ctcgtctgct gcctgatgca gttggaacaa cttgtggtca gcggcttgag aaggtgtatg 1920

gagccgagca ctcacatatt cttagggtcc cctttaggac cgagaagggc attgttcgca 1980

aatggatatc tcgctttgaa gtgtggccat acatggagac tttcactgag gtgacactaa 2040

gcttccttgt atttgtctat cttctaattg gtattaggaa caatttgcta attattaacg 2100

ctttggcttt tcgtacatca ggatgttgca aaagaacttg ctgcagaact gcaggccaag 2160

ccagatttga taattggcaa ctatagcgag ggaaatcttg tggcttcatt gctggctcac 2220

aagttaggcg taacgcaggt ctgtgttatt tttcacctct tataaatctg attgtatttc 2280

cattagtctg gaactaaaag tactaaaatt ttcttttctt cgctgtgtta tttgccttct 2340

gcagtgcacc attgcccatg cattggagaa aacaaagtat cctgattctg acatctactg 2400

gaaaaaattt gacgaaaaat accatttctc gtcccagttt accgctgatc ttattgcaat 2460

gaatcacacc gattttatca tcaccagcac tttccaggag atagcaggaa ggtataacat 2520

caattgctaa ttcggttgca gtaacatttt gttcgatttc ttccccttat gcttaaccta 2580

ataccctaat gaattttcca gcaaggacac tgtcggacag tacgagagtc accaggcatt 2640

cacaatgcct ggattgtaca gagtcgttca cggcattgat gtgttcgatc ccaaattcaa 2700

cattgtctca cctggagctg atataaacct gtatttccca tattccgaga aggaaaagag 2760

attgacagca cttcacccag aaattgagga gcttctgtac agtgatgttg agaacgagga 2820

acatctgtaa gtttctaact tactcgtacc gtcagtggca gagccagaat tttcattaaa 2880

atggggtcaa aatataaaga cataaattca caaagaagcc aaggggtgtc aatatgtagt 2940

ataaatatat taaaaaaatt acctagctac acaatgtaat tttccgacaa aggggtatcg 3000

gttgcacttc ttgaatacat gtggctctgc cactgggtac agttacaaag tcctgttacc 3060

tatgtagatg agcttgtgct gaacatgttg tgattttggt aggtgtgtgc taaaggacag 3120

gaataagcca atcttattca caatggcgag attggatcgt gtgaagaact taaccggact 3180

tgttgagtgg tacgccaaga acgcacggct aagggagttg gttaaccttg ttgtcgttgg 3240

tggagaccga aggaaggaat ccaaagattt ggaagagcaa gcagagatga agaagatgta 3300

tgagctaata aagactcaca acttaaatgg ccaattcaga tggatttctt cacagatgaa 3360

ccgagtaagg aacggcgaac tctaccgata cattgccgac actaggggag ctttcgtgca 3420

gcctgcattc tatgaggctt tcggtttgac tgttgttgag gccatgacct gtggtttgcc 3480

tacatttgca actaatcatg gcggtccagc tgagatcatc gttaacggaa aatccggctt 3540

ccatatcgat ccatatcacg gtgagcaagc tgctgatctg ctagctgatt tctttgagaa 3600

atgtaagacg gaaccttctc attgggaaac tatttcaacc ggtggcctga agcgcatcca 3660

agagaagtaa gcaactcttt cttgactcta gtcattcaaa ttaacttggg atttgaggca 3720

tagttgattg ataatttatc gcgtctctac tactatatac aggtacacgt ggcaaatcta 3780

ctcggagaga ttattgacgt tggctgctgt ttacggtttc tggaaacatg tttctaagct 3840

tgatcgtcta gaaatccgtc gatatctaga aatgttttat gctctcaaat accggaagat 3900

ggtgagttct tctgcttcct gctcttctca tagtgtttaa tatacacttg attgattgca 3960

ttcacttaga ctaagttgct cggacacggg tgtggatgtc cgacacgagt gcggatctag 4020

agttcagatc cttcaagatg taaattataa gattcgggga tatggatcct agtacggata 4080

cgggtgcgag aatccggcta aaaataattt taaaaaaaat tatctctaaa ttatgagata 4140

ttatgtggaa tacttacgta taacttgtaa agtgtagatt ttttttaatt ctcaagttgt 4200

agattagtaa atgattgatt tcctagataa gtatgctatt ttcttcaaat ttactcttct 4260

gatttcgaaa atcaaattgt atctcgtctc gaatttttcc gtccgttatg gtcaaagtac 4320

ccaaaatcgt ttgaccaaat cggtacggat cccataccca cacccacact agtgtcgtat 4380

tgacacgggt gccgcaccta aactgctatg tcggagcaac ttagcactta gagaatcatt 4440

gatgttaaat tttcttaatt cttgaatctg ctaatgaaga ttttatcttg gtttttgttt 4500

aggctgaagc tgttccatg gctgctgaat ga 4532

<210> 15

<211> 805

<212> PROTEIN

<213> Nicotiana tabacum

<400> 15

Met Ala Glu Arg Ala Leu Thr Arg Val His Ser Leu Arg Glu Arg Leu

1 5 10 15

Asp Ala Thr Leu Ala Ala His Arg Asn Glu Ile Leu Leu Phe Leu Ser

20 25 30

Arg Ile Glu Ser His Gly Lys Gly Ile Leu Lys Pro His Gln Leu Leu

35 40 45

Ala Glu Phe Asp Ala Ile Arg Gln Asp Asp Lys Lys Lys Leu Asn Asp

50 55 60

His Ala Phe Glu Glu Leu Leu Lys Ser Thr Gln Glu Ala Ile Val Leu

65 70 75 80

Pro Pro Trp Val Ala Leu Ala Ile Arg Leu Arg Pro Gly Val Trp Glu

85 90 95

Tyr Val Arg Val Asn Val Asn Ala Leu Val Val Glu Glu Leu Thr Val

100 105 110

Pro Glu Tyr Leu His Phe Lys Glu Glu Leu Val Asp Gly Thr Ser Asn

115 120 125

Gly Asn Phe Val Leu Glu Leu Asp Phe Glu Pro Phe Thr Ala Ser Phe

130 135 140

Pro Lys Pro Thr Leu Thr Lys Ser Ile Gly Asn Gly Val Glu Phe Leu

145 150 155 160

Asn Arg His Leu Ser Ala Lys Met Phe His Asp Lys Glu Ser Met Thr

165 170 175

Pro Leu Leu Glu Phe Leu Arg Val His Asn Tyr Lys Gly Lys Thr Met

180 185 190

Met Leu Asn Asp Arg Ile Gln Asn Leu Thr Thr Leu Gln Asn Val Leu

195 200 205

Arg Lys Ala Glu Glu Tyr Leu Ile Met Leu Pro Pro Glu Thr Pro Phe

210 215 220

Ser Glu Phe Glu His Lys Phe Gln Glu Ile Gly Leu Glu Lys Gly Trp

225 230 235 240

Gly Asp Thr Ala Glu Arg Val Leu Glu Met Ile Cys Met Leu Leu Asp

245 250 255

Leu Leu Glu Ala Pro Asp Ser Cys Thr Leu Glu Lys Phe Leu Gly Arg

260 265 270

Ile Pro Met Val Phe Asn Val Val Ile Leu Ser Pro His Gly Tyr Phe

275 280 285

Ala Gln Glu Asn Val Leu Gly Tyr Pro Asp Thr Gly Gly Gln Val Val

290 295 300

Tyr Ile Leu Asp Gln Val Pro Ala Leu Glu Arg Glu Met Leu Lys Arg

305 310 315 320

Leu Lys Glu Gln Gly Leu Asp Ile Thr Pro Arg Ile Leu Ile Val Thr

325 330 335

Arg Leu Leu Pro Asp Ala Val Gly Thr Thr Cys Gly Gln Arg Leu Glu

340 345 350

Lys Val Tyr Gly Ala Glu His Ser His Ile Leu Arg Val Pro Phe Arg

355 360 365

Thr Glu Lys Gly Ile Val Arg Lys Trp Ile Ser Arg Phe Glu Val Trp

370 375 380

Pro Tyr Met Glu Thr Phe Thr Glu Asp Val Ala Lys Glu Leu Ala Ala

385 390 395 400

Glu Leu Gln Ala Lys Pro Asp Leu Ile Ile Gly Asn Tyr Ser Glu Gly

405 410 415

Asn Leu Val Ala Ser Leu Leu Ala His Lys Leu Gly Val Thr Gln Cys

420 425 430

Thr Ile Ala His Ala Leu Glu Lys Thr Lys Tyr Pro Asp Ser Asp Ile

435 440 445

Tyr Trp Lys Lys Phe Asp Glu Lys Tyr His Phe Ser Ser Gln Phe Thr

450 455 460

Ala Asp Leu Ile Ala Met Asn His Thr Asp Phe Ile Ile Thr Ser Thr

465 470 475 480

Phe Gln Glu Ile Ala Gly Ser Lys Asp Thr Val Gly Gln Tyr Glu Ser

485 490 495

His Gln Ala Phe Thr Met Pro Gly Leu Tyr Arg Val Val His Gly Ile

500 505 510

Asp Val Phe Asp Pro Lys Phe Asn Ile Val Ser Pro Gly Ala Asp Ile

515 520 525

Asn Leu Tyr Phe Pro Tyr Ser Glu Lys Glu Lys Arg Leu Thr Ala Leu

530 535 540

His Pro Glu Ile Glu Glu Leu Leu Tyr Ser Asp Val Glu Asn Glu Glu

545 550 555 560

His Leu Cys Val Leu Lys Asp Arg Asn Lys Pro Ile Leu Phe Thr Met

565 570 575

Ala Arg Leu Asp Arg Val Lys Asn Leu Thr Gly Leu Val Glu Trp Tyr

580 585 590

Ala Lys Asn Ala Arg Leu Arg Glu Leu Val Asn Leu Val Val Val Gly

595 600 605

Gly Asp Arg Arg Lys Glu Ser Lys Asp Leu Glu Glu Gln Ala Glu Met

610 615 620

Lys Lys Met Tyr Glu Leu Ile Lys Thr His Asn Leu Asn Gly Gln Phe

625 630 635 640

Arg Trp Ile Ser Ser Gln Met Asn Arg Val Arg Asn Gly Glu Leu Tyr

645 650 655

Arg Tyr Ile Ala Asp Thr Arg Gly Ala Phe Val Gln Pro Ala Phe Tyr

660 665 670

Glu Ala Phe Gly Leu Thr Val Val Glu Ala Met Thr Cys Gly Leu Pro

675 680 685

Thr Phe Ala Thr Asn His Gly Gly Pro Ala Glu Ile Ile Val Asn Gly

690 695 700

Lys Ser Gly Phe His Ile Asp Pro Tyr His Gly Glu Gln Ala Ala Asp

705 710 715 720

Leu Leu Ala Asp Phe Phe Glu Lys Cys Lys Thr Glu Pro Ser His Trp

725 730 735

Glu Thr Ile Ser Thr Gly Gly Leu Lys Arg Ile Gln Glu Lys Tyr Thr

740 745 750

Trp Gln Ile Tyr Ser Glu Arg Leu Leu Thr Leu Ala Ala Val Tyr Gly

755 760 765

Phe Trp Lys His Val Ser Lys Leu Asp Arg Leu Glu Ile Arg Arg Tyr

770 775 780

Leu Glu Met Phe Tyr Ala Leu Lys Tyr Arg Lys Met Ala Glu Ala Val

785 790 795 800

Pro Leu Ala Ala Glu

805

<210> 16

<211> 5901

<212> DNA

<213> Nicotiana tabacum

<400> 16

atgcttttta tgggagtaaa ttttatggcc ggtcattcaa ctttgtgttc attacgcaaa 60

agtcattttt cttggtgttt attacgcaag tcatttttct tttttttttg ttacgtaaaa 120

atcattcaac tatgtgttta ttatctaaaa ttcaattttt tttttccttt tgttacacaa 180

aaatcatttt actttactct atttatcaca aaagtcacct tggccagatt ttataatagg 240

cttttatctt ttgttacaca aaaattattt tactttactc tatttatcac aaaagtcacc 300

ttggccagat tttataatag gcttttatct tttgttacac aaaaattatt ttactttact 360

ctatttatca caaaagtcac cttggccaga ttttacaata cttttacctt aaaagactat 420

tatgcccttg acattataaa tcctctcatt tatataatac cttctatatg atacactata 480

taatatattt ttacctaggt attttactta taattaaaat aatattaaat tattttattt 540

atctatttta taatatattc atacatttaa ttttttcatg gcaaatcact ttgtttaatc 600

atatttaaac atgaacaaat tttaaatatc aaaaaaataa aaaaataaaa aaaatattta 660

tttgaaataa taacaaacag atttgtttaa caaatgatag ttttttttta tagtcaataa 720

aatttttaaa aaaattcaaa gatatttgtt tttaatatta atatttttaa agctttatct 780

gttaatatta tttatttgaa agtattaatc tgatgtgtca ttgtgttaaa tgtgagtatt 840

ttatttattg gattaatgag tatggcttgg ctgataaaaa gctttgattt tataattttc 900

attaaaaata ttttattaag ctagtacctg acaaatttaa tatcttgaaa attaacgtta 960

agaaaaaatt aaatataaaa atatattata aaaataataa ataaataata tcaagttatt 1020

ttaattataa ataaaataca tggttaaaaa tatattatat agcatataat atagaaggta 1080

ttacataaat gagatgattt aaagggcata atagactttt caggtgaatg atttgtaaaa 1140

tatggttaaa gtgattattg tgataattag agcatagtaa aataattttt atgtaacaaa 1200

agaaaaaaaa aatgactttt gggtaatgaa cataaatttg aataactttt acgtaacaaa 1260

agaataaaat aaattttgga taataaacat aaaattgaat gaccacctat aaaatttatt 1320

atttttttgg gctcttcttg atttgatttt ttagtttagc ctttgcagta atcttggttg 1380

tcacgcgtag cgttgtgctt tcgccacata agtatttagt agacttaatt aatgtcatta 1440

tatcggttgg tgtggtttta attacttaac tgtactatta tattaggtgg aaggtttgaa 1500

aatttatagt agtaacattc tagatcattg aaaatattgg tgtttcagtg actttttagt 1560

atgtcatttt cattttctaa gtggttgtac taatatagta tattaaaatt ttgattggtt 1620

gagaaacaat ctctctcacc tacacggtac gggtaaggta tgcgtatacg cttatcctcc 1680

ctacactcca tttgtgggac tattgttgtt attttggata agctgaggta tccatcttct 1740

actaactgca ctagtttatt ttttttgctg tttacagttg aaacaattgt ctgaggattt 1800

ctcacctgct gaatcaactg caatggctga acgtgtgctg actcgtgttc acagccttcg 1860

tgaacgtctt gatgctactt tggctgctca tcgcaatgag atattactgt ttctttcaag 1920

gtatagccaa agatagtatt cttgttaact aaaaaagatt cagttggtgt tcaaaaaacg 1980

atacgtttat ctgcctaagt cttggtagtc agaattatcc ggtacctatg ctggtgtgag 2040

ttagctggct aggaaaccac tcttatgaaa acaagagatt tagttagagt tgtctgtaat 2100

tctgtagtat ggactatgta tgtgatgcta tttgaactgg ttttggttat tataggattg 2160

aaagccatgg aaaagggatc ttgaaaccgc atcagctatt ggctgagttt gatgcaattc 2220

gccaagatga caaaaagaaa ctgaatgatc atgcatttga agaactcctg aagtccactc 2280

aggtaatatg gttttggcta tatttgtcgc caacgccaag ctcatatttt tatattattt 2340

tgagcttgtg tctgaatacg acgatgatat gttatactag gaagcaattg ttctgccacc 2400

ttgggttgca cttgcgattc gtttgaggcc tggtgtgtgg gaatatgtcc gtgtgaatgt 2460

caatgcgcta gtcgttgagg agctgactgt ccctgagtat ttgcatttca aggaagaact 2520

tgtcgatgga acgtaagtgt tagtcttcaa tttgatgcta tgttagagaa taggctgtgg 2580

aatttattga tcaatgctgt gctttgtcct gatacagctc caatggaaat ttcgttctcg 2640

agttggattt tgagcccttc accgcatcct ttcctaaacc aaccctcacc aaatctatcg 2700

gaaatggagt tgaattcctc aataggcacc tctctgcgaa aatgttccat gacaaggaaa 2760

gcatgacccc gcttcttgaa tttcttcggg ttcacaatta taagggcaag gtgacttgct 2820

atttccatt atctataggt tcggtttgtg cttatcatgc gcccaatgac atatgaatat 2880

gcgctaaagg atagatatat gatttccttt gcagacaatg atgctgaacg acagaataca 2940

gaatttaacc acactgcaaa atgtcctaag gaaggcagag gaatacctca ttatgcttcc 3000

ccctgaaact ccattttccg aattcgaaca caagttccaa gaaattggat tggagaaggg 3060

atggggcgac actgcagagc gcgtgctgga gatgatatgc atgcttcttg atctcctcga 3120

ggctcccgat tcctgtactc ttgagaagtt cttggggaga attcctatgg tgttcaatgt 3180

ggttatcctt tccccccacg gatatttcgc cccaggaaaat gtcttgggtt atcccgacac 3240

tggtggccag gtgcattact ttaatcttta tccgtgagtc tatgtttgtt cgaatcctct 3300

agaaatgtca ctgtacctat gtaggatact ccaaatataa tgcattttgg ggggatctgt 3360

tatgggtgcg atggcatttt tggaggtcgg agcaacaaac aattgctatg tattcttcta 3420

aagcttgctt tcataaatgc taaggtcctt cacccttaat gtgcaggttg tctatatatt 3480

agatcaagtt ccagccttgg agcgtgaaat gcttaagcgc ctaaaggagc aaggacttga 3540

tatcacaccg cgtattctta ttgttagtat ttcctgtact tgtaattact gcggattaca 3600

caaaatttcc tttttatctt cttaacaact tatcttgatg gtattcccag gttactcgtc 3660

tgctacctga tgcagttgga acgacttgtg gtcagcggct tgagaaggtg tatggagccg 3720

agcactcaca tattctgagg gtccccttta ggactgagaa gggcattgtt cgtaaatgga 3780

tctctcgctt tgaagtgtgg ccatatatgg agactttcac tgaggtgaca ctaaaacttc 3840

cttatatttg tctatcttct aattggtatt aggaataatt tgttaattgt taactctttg 3900

tcttttcgta catcaggatg tcgcaaaaga acttgctgca gaattgcagg ccaagccaga 3960

tttgataata ggcaactata gcgagggaaa tcttgtggct tcattgctcg ctcataagtt 4020

aggcgtaaca caggtctgtg ttgtttttca ctctcttaaa gatctgattg catttccatt 4080

agtctggaac tagaagtact aaaaagttct tttcttcact gtgttatttg ccgtcggcag 4140

tgcaccatag ctcatgcatt ggagaaaaca aagtatcctg attctgacat ctactggaaa 4200

aaattcgatg aaaaatacca tttctcgtcc cagtttaccg ctgatcttat tgcaatgaat 4260

cacaccgatt ttatcatcac cagcactttc caggagatag caggaaggta taacatcaat 4320

ttgctacttc gactgcaaca gcattgtgtt cccatttctt tcccttatgc ttaacctaat 4380

accgtcatga attttccagc aaggacactg tcggacagta cgagagtcat caggcattca 4440

caatgcccgg attgtacaga gttgttcacg gcattgatgt gttcgacccc aaattcaaca 4500

ttgtctcacc tggagctgac ataaacctct atttcccata ttccgagaag gaaaagagac 4560

tgacagcact tcaccctgaa atcgaggagc tgctgtacag tgacattgag aacgaggaac 4620

atctgtaagt ttctacctta ctcgtacagt cagtggcgga gccagaattt tcactaaaat 4680

aaggtcaaaa tataaagaca taaatccaca aagaagccaa gggtgtcaat atatagtata 4740

aatacattaa aaaaattacc tatctacaca gtgtaatttt ccgacaaagg ggtgtcggtt 4800

gacactcctt gaatacatgt ggctctgcca ctgggtacag ttacaaagtt ctgttaccta 4860

tgtagatgag cttgtgctga acatgttgtg attttggcag gtgtgtgcta aaggacagga 4920

ataagccaat cttattcaca atggcgagat tggatcgtgt gaagaattta accggacttg 4980

ttgagtggta tgccaagaac gcacggctaa gggagttggt taaccttgtt gtggttggtg 5040

gagatcgaag gaaagaatcc aaagatttgg aagagcaaac agaaatgaaa aagatgtatg 5100

agctaataaa gactcacaat ttaaatggcc aattcagatg gatttcttca cagatgaacc 5160

gagtgaggaa cggtgaactc taccgataca ttgctgacac tagagggagct ttcgtgcagc 5220

ctgcattcta cgaggctttc ggtttgactg ttgttgaggc catgacctgt ggtttgccta 5280

catttgcaac taatcatggc ggtccagctg agatcatcgt taacggaaaa tctggcttcc 5340

acatcgatcc atatcacggt gagcaagctg ctgatctgct agctgatttc tttgagaaat 5400

gtaagacaga accttctcat tgggaaacca tttcaacggg tggcctgaag cgcatccaag 5460

agaagtaagc aactctttct tgactctagt cattgaaatt aactttcttg actctagtca 5520

ttgaaattaa ctcgggattt gaggcgtagt tgattgatat tttatcgcgt ctctactact 5580

gatatataca ggtacacgtg gcaaatctac tcggagaggc tattgacatt ggctgctgtt 5640

tacgggttct ggaaacatgt ttctaagctt gatcgtctag aaatccgtcg atatcttgaa 5700

atgttttatg ctctcaaata ccgcaagatg gtgagttcct cttcttcctt gcccttctcc 5760

tagtgtttaa gatacaatat aattgattgc attatcttag agaatcatta atgttaaatt 5820

ttcttaattc ttgaatctgt taatgaagtt tttctcttgg tttttgttta ggctgaagct 5880

gttccattgg ctgctgagtg a 5901

<210> 17

<211> 825

<212> PROTEIN

<213> Nicotiana tabacum

<400> 17

Met Leu Phe Met Gly Leu Lys Gln Leu Ser Glu Asp Phe Ser Pro Ala

1 5 10 15

Glu Ser Thr Ala Met Ala Glu Arg Val Leu Thr Arg Val His Ser Leu

20 25 30

Arg Glu Arg Leu Asp Ala Thr Leu Ala Ala His Arg Asn Glu Ile Leu

35 40 45

Leu Phe Leu Ser Arg Ile Glu Ser His Gly Lys Gly Ile Leu Lys Pro

50 55 60

His Gln Leu Leu Ala Glu Phe Asp Ala Ile Arg Gln Asp Asp Lys Lys

65 70 75 80

Lys Leu Asn Asp His Ala Phe Glu Glu Leu Leu Lys Ser Thr Gln Glu

85 90 95

Ala Ile Val Leu Pro Pro Trp Val Ala Leu Ala Ile Arg Leu Arg Pro

100 105 110

Gly Val Trp Glu Tyr Val Arg Val Asn Val Asn Ala Leu Val Val Glu

115 120 125

Glu Leu Thr Val Pro Glu Tyr Leu His Phe Lys Glu Glu Leu Val Asp

130 135 140

Gly Thr Ser Asn Gly Asn Phe Val Leu Glu Leu Asp Phe Glu Pro Phe

145 150 155 160

Thr Ala Ser Phe Pro Lys Pro Thr Leu Thr Lys Ser Ile Gly Asn Gly

165 170 175

Val Glu Phe Leu Asn Arg His Leu Ser Ala Lys Met Phe His Asp Lys

180 185 190

Glu Ser Met Thr Pro Leu Leu Glu Phe Leu Arg Val His Asn Tyr Lys

195 200 205

Gly Lys Thr Met Met Leu Asn Asp Arg Ile Gln Asn Leu Thr Thr Leu

210 215 220

Gln Asn Val Leu Arg Lys Ala Glu Glu Tyr Leu Ile Met Leu Pro Pro

225 230 235 240

Glu Thr Pro Phe Ser Glu Phe Glu His Lys Phe Gln Glu Ile Gly Leu

245 250 255

Glu Lys Gly Trp Gly Asp Thr Ala Glu Arg Val Leu Glu Met Ile Cys

260 265 270

Met Leu Leu Asp Leu Leu Glu Ala Pro Asp Ser Cys Thr Leu Glu Lys

275 280 285

Phe Leu Gly Arg Ile Pro Met Val Phe Asn Val Val Ile Leu Ser Pro

290 295 300

His Gly Tyr Phe Ala Gln Glu Asn Val Leu Gly Tyr Pro Asp Thr Gly

305 310 315 320

Gly Gln Val Val Tyr Ile Leu Asp Gln Val Pro Ala Leu Glu Arg Glu

325 330 335

Met Leu Lys Arg Leu Lys Glu Gln Gly Leu Asp Ile Thr Pro Arg Ile

340 345 350

Leu Ile Val Thr Arg Leu Leu Pro Asp Ala Val Gly Thr Thr Cys Gly

355 360 365

Gln Arg Leu Glu Lys Val Tyr Gly Ala Glu His Ser His Ile Leu Arg

370 375 380

Val Pro Phe Arg Thr Glu Lys Gly Ile Val Arg Lys Trp Ile Ser Arg

385 390 395 400

Phe Glu Val Trp Pro Tyr Met Glu Thr Phe Thr Glu Asp Val Ala Lys

405 410 415

Glu Leu Ala Ala Glu Leu Gln Ala Lys Pro Asp Leu Ile Ile Gly Asn

420 425 430

Tyr Ser Glu Gly Asn Leu Val Ala Ser Leu Leu Ala His Lys Leu Gly

435 440 445

Val Thr Gln Cys Thr Ile Ala His Ala Leu Glu Lys Thr Lys Tyr Pro

450 455 460

Asp Ser Asp Ile Tyr Trp Lys Lys Phe Asp Glu Lys Tyr His Phe Ser

465 470 475 480

Ser Gln Phe Thr Ala Asp Leu Ile Ala Met Asn His Thr Asp Phe Ile

485 490 495

Ile Thr Ser Thr Phe Gln Glu Ile Ala Gly Ser Lys Asp Thr Val Gly

500 505 510

Gln Tyr Glu Ser His Gln Ala Phe Thr Met Pro Gly Leu Tyr Arg Val

515 520 525

Val His Gly Ile Asp Val Phe Asp Pro Lys Phe Asn Ile Val Ser Pro

530 535 540

Gly Ala Asp Ile Asn Leu Tyr Phe Pro Tyr Ser Glu Lys Glu Lys Arg

545 550 555 560

Leu Thr Ala Leu His Pro Glu Ile Glu Glu Leu Leu Tyr Ser Asp Ile

565 570 575

Glu Asn Glu Glu His Leu Cys Val Leu Lys Asp Arg Asn Lys Pro Ile

580 585 590

Leu Phe Thr Met Ala Arg Leu Asp Arg Val Lys Asn Leu Thr Gly Leu

595 600 605

Val Glu Trp Tyr Ala Lys Asn Ala Arg Leu Arg Glu Leu Val Asn Leu

610 615 620

Val Val Val Gly Gly Asp Arg Arg Lys Glu Ser Lys Asp Leu Glu Glu

625 630 635 640

Gln Thr Glu Met Lys Lys Met Tyr Glu Leu Ile Lys Thr His Asn Leu

645 650 655

Asn Gly Gln Phe Arg Trp Ile Ser Ser Gln Met Asn Arg Val Arg Asn

660 665 670

Gly Glu Leu Tyr Arg Tyr Ile Ala Asp Thr Arg Gly Ala Phe Val Gln

675 680 685

Pro Ala Phe Tyr Glu Ala Phe Gly Leu Thr Val Val Glu Ala Met Thr

690 695 700

Cys Gly Leu Pro Thr Phe Ala Thr Asn His Gly Gly Pro Ala Glu Ile

705 710 715 720

Ile Val Asn Gly Lys Ser Gly Phe His Ile Asp Pro Tyr His Gly Glu

725 730 735

Gln Ala Ala Asp Leu Leu Ala Asp Phe Phe Glu Lys Cys Lys Thr Glu

740 745 750

Pro Ser His Trp Glu Thr Ile Ser Thr Gly Gly Leu Lys Arg Ile Gln

755 760 765

Glu Lys Tyr Thr Trp Gln Ile Tyr Ser Glu Arg Leu Leu Thr Leu Ala

770 775 780

Ala Val Tyr Gly Phe Trp Lys His Val Ser Lys Leu Asp Arg Leu Glu

785 790 795 800

Ile Arg Arg Tyr Leu Glu Met Phe Tyr Ala Leu Lys Tyr Arg Lys Met

805 810 815

Ala Glu Ala Val Pro Leu Ala Ala Glu

820 825

<210> 18

<211> 8323

<212> DNA

<213> Nicotiana tabacum

<400> 18

atggcgaatc caaagttcac aagagtacct agcatgaggg agagagttga ggatactctc 60

tctgctcacc gtaaccagct tgttgctctc ctctccaggt atattaataa actctatata 120

cttgttattt tctttatttt tttgtcttta ctgataaatt taactgtttt cttctttaaa 180

tcttgctttc gatgcatgat ttctgttgtg ttaaattgcg taaccatttt atctaaaagt 240

ttatgctgat aaacactttt aaattttaat atgtaaatta tattatgtct caacatcaac 300

atgtggatgg ccaaaaatat aaagcttaat tttcgttatt ttgaatgatt tttctctgcg 360

agtgttacgg tttgcgtaca cattacctaa acctcctccc tagtccccac ttgtgggaat 420

ttaatttttt ttttctttgt ttttttttgt tgttgttgtt gtctgagttc aattcctacc 480

atgttagctt ggcaaaaata agttggtaaa gcttgacccc aactagtttt agttgatcga 540

tttatttggt gatttatagt tcaataataa taattactat tagagaaagt tccagcagct 600

tttctgtttg tttttccagt tttagtgatt gatatatgtg tatatatatt ctttgtttct 660

tttaagatac gtggcgcagg ggaaggggat attgcaacct caccacttga tcgatgagtt 720

caacaacgct gtatgtgatg acactgcttg tgagaagctc aaagatggtc cctttagtga 780

agtcttgaaa gctactcagg tatattcact aatccatggg aatcaagatg atactgtata 840

tctttattat ggtgtctttc agaaatttga cgatgatgaa atgcaacttt tctctgtttg 900

tcaccttatc cagactgttt ttttattttt tatttttcat tttttaactt gaaatgctct 960

taatttcctt tgtttatcga taagaccgga tttacaatgt atgaacggag catcttaaga 1020

accttctgga atgaagatat aagatataaa acatggtgtc cgttttctcc tttgtggaat 1080

cagtgtacat atagactgtt attttggtcc cactttctgg atcttctgat cacaccttct 1140

catgcagagg cgagcttgat ggtttcaacc tttaaattct tactattgaa tccatttcac 1200

tttcgaaatt atgagttcga aatctaatat ttgttgaaat ttttgcaaat gttcacatat 1260

aagtttaagc tttgtgtcaa gaatactggg ctcaatggat tccaatagac caggctgtat 1320

ccgcctctgt ctccactctc cctgcatcca cttctttcgt gtgactaata atgcttaatg 1380

agctagaact cgttttaatg tttgaataag ttgcttatat cagagcagct tttgatgttt 1440

caatctttaa cgggttatgc agtaccagca ttctgcggct gaaaaacagg aatctgagat 1500

ttacttgtct ctggctgaat ttcttgttca ttttgctaac aagtactttg gagttaatgc 1560

ttgctctctg ttgtcaaaat aggaagccat tgtgctgcca ccatttgttg ccatagcagt 1620

tcgtccaagg ccaggtgttt gggagtatgt tcgtgttaat gtatatgatt tgagcgttga 1680

acaattgact gttcctgaat atcttcattt caaggaagaa cttgtggatg gagagtaagc 1740

tctttcttat ttcaatacga aacataaaaa tttacagaag ttgaataatt aacaaatttg 1800

ttgattttta atgtatgcca ggggtaataa tcactttgtg cttgagctgg attttgagcc 1860

atttaatgca tcagttcctc gtccatctcg atcgtcatcc attggcaatg gagtccaatt 1920

cctcaatcgt catctttcct caattatgtt tcgcagcaaa gactctctgg accccttact 1980

tgatttcctt agaggacact gtcataaagg gaatgtaagt accaaaagca gttttccctt 2040

tgtaaatgtc tgcttgtccc tgattatcta ctaaatcttt caacacgcgc aaccattata 2100

agaaatgtac aatacttcta gttagaattt catcatcgac aaactatctg ctttactttt 2160

tatttttccc atttgatgga tgatagttta gtttatataa cagatgatat tttggttgaa 2220

gggtaccatg aactttttca caaccactta atggatacat agttgtaata gttgacattt 2280

tggaataata ttgtctcact tggaaatgtt taagaagtat tactacttct atttgtaaga 2340

tggattgttt atctatgcag gtcttgatgt tgaatgatcg tatacagcga atctccaggc 2400

tggagtctgc tctttctaaa gcagaggatt atctctccaa gctatcacca gatacatcct 2460

ataatgagtt cgaatacgcg tgagcttgta cacatttgtt ttgttttctt tcaagcatat 2520

gtaatttctc aagaaaaggg aaatctatag gagttgaaac attctttatg gaaccatgtg 2580

catgcagatt gcaagaaatg ggctttgaga gaggttgggg tgatactgcc agacgtgttt 2640

tggagacgat gcatcttctt tctgacattc ttcaggctcc ggatccatca accttggaga 2700

catttcttgg tagactacct atggtgttca atgtcgtcat attatcccct catggatatt 2760

ttggccaagc aaatgtcttg ggtttgcccg acactggtgg ccaggtaata acaaggagaa 2820

tgaggtcttg tattatgtac tccctccgtt ccaatctata tgaacctatt tgactgggta 2880

tggaaagaaa tgaagacttg taaaacttgt ggttctttag aaattccaaa cattacattt 2940

ggttttttcc ctcttcctgg aaattatact actgaatcat ctctagatgt tccagtttaa 3000

cttgagacgt aagggtaaat aacggaccat tactctgtcc tttcttgcag taggcttggt 3060

acaatgaata tagttcgcat agttgccgga agctagagct gtgttagaaa actcaggaac 3120

attaatttgg cgatgctaat cactgctaat gttactgaag catccatggt tttccttgat 3180

gttattctcc ttttggttgc ttcacaggtt gtctatatac tggatcaagt gcgtgccttg 3240

gaggccgaaa tgcttcttag aataaagcaa caaggactta acttcaagcc tagaatcctt 3300

gtcgtgagta catatatatt atgcaagctc ttatttggtt tgtgggattg cagttgacat 3360

caatttgctt actctgatta ctaaaggtca cacggctgat acctgatgct aaaggaacca 3420

tgtgcaacca gaggttggag aggattagtg gaactgaata ctcgcatatt ttacgtgtcc 3480

cttttaggac agagaaggga atccttcata aatggatatc taggtttgat gtatggcctt 3540

acctggagaa gttcactgag gtaacctctt tgtcccttgg aaattgcctt ttgttgctga 3600

tgtttctgct agtgtgctta aatgacggat gttaactagt cacttgctag cgtttgcaat 3660

agcaacggga aaagaaagga tttttgctag tttgaagtct gcctccaaga aaaattatat 3720

taaaagttta tggctagtgg aaacatcagt cattcatgta ccttatttct atgcccaagt 3780

tgtttaagtt gaaagtaatt tggccaacta tgcaaattgg gagaacgtgt agccaactat 3840

tgtgtttgcc gacatgttga tatacttttt ggtcctgatt tatatttgtt ggtttgtcat 3900

actggatgaa gcaattctca tgtttttctg cttatatata ttggaagaag agatacttgt 3960

cgtttcatca tttttctcga cctctctatt accaacactt tgccaattta atgtttggaa 4020

atgtcttctt gaccaggatg tggcaagtga aatgaccgct gagctccagg gaaagccaga 4080

tctgattatt ggcaactaca gtgatggaaa tttagttgcc tcccttttgg catataaaat 4140

gggtgtcaca caggtaggaa atacatgatt ctttatcttg ctagcactaa gtcttgaggt 4200

tatgtatctg caatagaaat tttacgcttt gccttcattt ctttttaatt atttttccag 4260

tgtaccattg ctcatgcctt ggaaaaaaca aagtatcctg attctgacat ctactggaaa 4320

aagtttgagg agaaatatca tttttcatgt cagtttactg ctgatctact ggcaatgaat 4380

aattcagatt tcattatcac cagtacttat caagagattg caggaacgta agtcatttta 4440

atctggtcgt ttaaatctga tatttcttcc ctagtagtct attcaatccg aatttcagtt 4500

cagtatatga tgtcatcggt tgaggaactg tgattggtaa ccttatcaaa tccgtagctg 4560

ctctataatt ttatttcgta attggagaaa caatttttta ttattgagct tgtagtctga 4620

gctagaattt ggttctttat ctatcaagta gcataatact acaactattt tttatgtgtg 4680

gcaatttgca atttcaattt tctatttcta taagttgcag cttttcttcc tgttctgatc 4740

atatttacat ggctgaaact caatagaaaa ctaggctagt tgatcaaaag tagttggatg 4800

ctttaaaatt agtagacgtt ttgctaaatg agtgaccaat gttattaaaa aaacgttcat 4860

gttttcaacc cttttggcat acatttgacc actgcccaag attttggata agtacatgca 4920

gtgcttataa ttataaagca ttttatccca ccttgttttt cattatgaaa attaagtaat 4980

ttacgagtat ttgtataagt tacttcataa attagaagta aatctggatt gtgtaaagtt 5040

attcgccccg tatatactga aagctacttg aacaagcaaa aaaacagaca aacgtaacat 5100

tctccatgga ttaatgagac ttgtatatat atatatatat atatgtaaag agagagagag 5160

agagatttgg cttgtaacca catgtatatt atgccatatg gatgtgacat tgatgtgact 5220

agacctaaat gttttgtttc aatgtccacg ggagttttac gtagagttaa gaggagaaga 5280

gagtgaggaa tactaatgtt tgatggtacc ccttggcttc ttgacctgga tactcagtgt 5340

tcttattcat gcctatactt tggtccttga tttcattctc ccttttctag cttgagctgc 5400

atcaaagaaa ttccactgta aaaaaaataa tgctcaccat attggtgcaa catggcaaac 5460

atgtatccta tttgatgatc aatcaacttt atttttctcc tgttaattga cctcagtgtg 5520

taactctcta tgtatgatag cattgtaact tgtgtcatga ttcataaata gggtactaga 5580

attggatggt tgacatagta aatggtcaat tgatgatcca caaaatatgc acctactgat 5640

taaaatgtga tagggcaggt ttatttttgt ttgtggttaa cacagtactt aaccctatat 5700

ttaatacaat ttggcttatc tacaatcttt tcttcagtgt ttatgcgaat tccttattgc 5760

acaacaatat tgtctttctg agttctattc tgttgttgct tacactttta ttattccagt 5820

aacatagatg tgaagacatt agattggttg cttgcaaatt gatagccact tgtttcagga 5880

agaatactgt tggtcagtac gagagccata ctgcattcac cctcccggga ctatatcgcg 5940

tcgttcatgg cattgatgtt ttcgatccca aattcaatat agtgtctcct ggagctgaca 6000

tgacaattta tttcccatat tctgacaagg aaaaaagact aacgtctttg catggctcga 6060

ttgaaaagtt gttatttgat cctgcgcaga atgaagagca tatgtaagtg gcatccgttt 6120

gtacttaatt tttttggaat agatgacata ttatttgcat gaatatgaaa aggagggtct 6180

gatatgattt tctatagata aactaccaat gatattattt aaaaactcct ggatactgta 6240

ttaggagaag aagagaacca ggggtagatg gcattagaat cccttaaatc ttgaagagtc 6300

gtcactaacg ctcccaacac ttctgcctca gaccctcaac taaatactat tattgttgat 6360

ttctttggag aagctataag aatctctctc tccttatggt gaaaatttta cttggcttta 6420

tacttaactt ccaaggctcc ctcttataaa atgcaaaaac tgtctgtatt cactctcttg 6480

gttaacaatt gatccaatca aatgcatatg gaacatcttt ctttacgttt cttctaaagt 6540

tcgtttgagg ataaggagta gaatctgaga agatagacta gtaggtaacc ttagggacgg 6600

atgtggaaat taacatatgg gctcagcttt tctgccgagt gcagaccatg tatatgcgtt 6660

aaaaaattca ctaaacaagt aaatgtttga ttttgaaccc agtaaatcaa atgagttgtg 6720

gtagaatctc gaactcgaac cgataaagtt caaatccagg atccgctttt aggtaaactc 6780

taccttggga agtgttatat atatgtccct gattatttct ttttccgttt cctttctatt 6840

ttaattttta aagttatttt tagatggttt tattttttga taagtggtaa gttgttaata 6900

ttccaaatta aatgccattg tcataactat atacatttat aaagaatgat tgatcctagt 6960

ttctcattcc taagatccaa ataaggcaat aaacaatgtc ttagtaattg gacctgcttc 7020

tggtgatcaa cgcttgatcg cgtagttagt tatagatgac tgtaaaaact ttaaccattt 7080

taatggtttt gtcaaagaac aaatatcgga catattatag agaatggact attgtacttt 7140

gcttctgatt ggtcatttta ttgtgatccg taaattggct gtgactgatg tcatatcttt 7200

gcttacagag gtaatctgaa tgataaatca aaacccataa ttttttcaat ggcaaggcta 7260

gaccatgtta agaacattac gggactagtt gagtgctatg ctaaaaatgc cacattgagg 7320

gaattggcga accttgttgt agtagctgga tacaacgatg taaagaaatc cagtgataga 7380

gaagaaataa cagaaattga gaagatgcat gctcttatta aggagcataa attggatggg 7440

caattcagat gggtatcagc ccaaacaaac cgggcacgta atggtgagct ctatcgctat 7500

atagctgacc agagaggtat atttgttcag gtatgctatt tgtattgtat tagtccaatt 7560

tcattttttg caccaaaaga aaggttgtta ttgtgacgta tatgtttgtt ttagcctgca 7620

ttttatgaag catttggact aacggtggtt gaagctatga cttgtggtct tccaacattt 7680

gcaacttgcc atggtggtcc taatgagatc attgaacccg gtgtatctgg gttccatatt 7740

gatccttatc atcccgataa agctgctgaa ctcatgtcag aattctttca acgctgcaaa 7800

caagatccta ctcactggga aaaaatatct gcatctggtc tccgaaggat tcttgagagg 7860

tctgtagttg tgtacatgta tagaagatta aagaatgcta ccttgatatt tatttgaatc 7920

aaaaataaca ggaacatctc ttttttgaac atcactcaag ttcttatatt aaataatttt 7980

taggtatacg tggaagattt actccgagag gctgatgact ttatctggcg tatatggttt 8040

ctggaagctt gtttcaaaac ttgagaggcg tgaaactaga cgataccttg agatgttcta 8100

cattctcaaa ttccgcgagt tggtgagtgc cttttagctc cttttcagtt ccaataaact 8160

atatatgtgg tttaagtaag tattaagcat aaacatgtcc gtgcttgggg ctgtcgaaaa 8220

tgctatggac atatcctgag ctaaggattt ttcaagaaaa ttgatgttag ctttactcta 8280

tttacaggca aaatctgtac ctctagcaat tgatgacaag tga 8323

<210> 19

<211> 810

<212> PROTEIN

<213> Nicotiana tabacum

<400> 19

Met Ala Asn Pro Lys Phe Thr Arg Val Pro Ser Met Arg Glu Arg Val

1 5 10 15

Glu Asp Thr Leu Ser Ala His Arg Asn Gln Leu Val Ala Leu Leu Ser

20 25 30

Arg Tyr Val Ala Gln Gly Lys Gly Ile Leu Gln Pro His His Leu Ile

35 40 45

Asp Glu Phe Asn Asn Ala Val Cys Asp Asp Thr Ala Cys Glu Lys Leu

50 55 60

Lys Asp Gly Pro Phe Ser Glu Val Leu Lys Ala Thr Gln Glu Ala Ile

65 70 75 80

Val Leu Pro Pro Phe Val Ala Ile Ala Val Arg Pro Arg Pro Gly Val

85 90 95

Trp Glu Tyr Val Arg Val Asn Val Tyr Asp Leu Ser Val Glu Gln Leu

100 105 110

Thr Val Pro Glu Tyr Leu His Phe Lys Glu Glu Leu Val Asp Gly Glu

115 120 125

Gly Asn Asn His Phe Val Leu Glu Leu Asp Phe Glu Pro Phe Asn Ala

130 135 140

Ser Val Pro Arg Pro Ser Arg Ser Ser Ser Ile Gly Asn Gly Val Gln

145 150 155 160

Phe Leu Asn Arg His Leu Ser Ser Ile Met Phe Arg Ser Lys Asp Ser

165 170 175

Leu Asp Pro Leu Leu Asp Phe Leu Arg Gly His Cys His Lys Gly Asn

180 185 190

Val Leu Met Leu Asn Asp Arg Ile Gln Arg Ile Ser Arg Leu Glu Ser

195 200 205

Ala Leu Ser Lys Ala Glu Asp Tyr Leu Ser Lys Leu Ser Pro Asp Thr

210 215 220

Ser Tyr Asn Glu Phe Glu Tyr Ala Leu Gln Glu Met Gly Phe Glu Arg

225 230 235 240

Gly Trp Gly Asp Thr Ala Arg Arg Val Leu Glu Thr Met His Leu Leu

245 250 255

Ser Asp Ile Leu Gln Ala Pro Asp Pro Ser Thr Leu Glu Thr Phe Leu

260 265 270

Gly Arg Leu Pro Met Val Phe Asn Val Val Ile Leu Ser Pro His Gly

275 280 285

Tyr Phe Gly Gln Ala Asn Val Leu Gly Leu Pro Asp Thr Gly Gly Gln

290 295 300

Val Val Tyr Ile Leu Asp Gln Val Arg Ala Leu Glu Ala Glu Met Leu

305 310 315 320

Leu Arg Ile Lys Gln Gln Gly Leu Asn Phe Lys Pro Arg Ile Leu Val

325 330 335

Val Thr Arg Leu Ile Pro Asp Ala Lys Gly Thr Met Cys Asn Gln Arg

340 345 350

Leu Glu Arg Ile Ser Gly Thr Glu Tyr Ser His Ile Leu Arg Val Pro

355 360 365

Phe Arg Thr Glu Lys Gly Ile Leu His Lys Trp Ile Ser Arg Phe Asp

370 375 380

Val Trp Pro Tyr Leu Glu Lys Phe Thr Glu Asp Val Ala Ser Glu Met

385 390 395 400

Thr Ala Glu Leu Gln Gly Lys Pro Asp Leu Ile Ile Gly Asn Tyr Ser

405 410 415

Asp Gly Asn Leu Val Ala Ser Leu Leu Ala Tyr Lys Met Gly Val Thr

420 425 430

Gln Cys Thr Ile Ala His Ala Leu Glu Lys Thr Lys Tyr Pro Asp Ser

435 440 445

Asp Ile Tyr Trp Lys Lys Phe Glu Glu Lys Tyr His Phe Ser Cys Gln

450 455 460

Phe Thr Ala Asp Leu Leu Ala Met Asn Asn Ser Asp Phe Ile Ile Thr

465 470 475 480

Ser Thr Tyr Gln Glu Ile Ala Gly Thr Lys Asn Thr Val Gly Gln Tyr

485 490 495

Glu Ser His Thr Ala Phe Thr Leu Pro Gly Leu Tyr Arg Val Val His

500 505 510

Gly Ile Asp Val Phe Asp Pro Lys Phe Asn Ile Val Ser Pro Gly Ala

515 520 525

Asp Met Thr Ile Tyr Phe Pro Tyr Ser Asp Lys Glu Lys Arg Leu Thr

530 535 540

Ser Leu His Gly Ser Ile Glu Lys Leu Leu Phe Asp Pro Ala Gln Asn

545 550 555 560

Glu Glu His Ile Gly Asn Leu Asn Asp Lys Ser Lys Pro Ile Ile Phe

565 570 575

Ser Met Ala Arg Leu Asp His Val Lys Asn Ile Thr Gly Leu Val Glu

580 585 590

Cys Tyr Ala Lys Asn Ala Thr Leu Arg Glu Leu Ala Asn Leu Val Val

595 600 605

Val Ala Gly Tyr Asn Asp Val Lys Lys Ser Ser Asp Arg Glu Glu Ile

610 615 620

Thr Glu Ile Glu Lys Met His Ala Leu Ile Lys Glu His Lys Leu Asp

625 630 635 640

Gly Gln Phe Arg Trp Val Ser Ala Gln Thr Asn Arg Ala Arg Asn Gly

645 650 655

Glu Leu Tyr Arg Tyr Ile Ala Asp Gln Arg Gly Ile Phe Val Gln Pro

660 665 670

Ala Phe Tyr Glu Ala Phe Gly Leu Thr Val Val Glu Ala Met Thr Cys

675 680 685

Gly Leu Pro Thr Phe Ala Thr Cys His Gly Gly Pro Asn Glu Ile Ile

690 695 700

Glu Pro Gly Val Ser Gly Phe His Ile Asp Pro Tyr His Pro Asp Lys

705 710 715 720

Ala Ala Glu Leu Met Ser Glu Phe Phe Gln Arg Cys Lys Gln Asp Pro

725 730 735

Thr His Trp Glu Lys Ile Ser Ala Ser Gly Leu Arg Arg Ile Leu Glu

740 745 750

Arg Tyr Thr Trp Lys Ile Tyr Ser Glu Arg Leu Met Thr Leu Ser Gly

755 760 765

Val Tyr Gly Phe Trp Lys Leu Val Ser Lys Leu Glu Arg Arg Glu Thr

770 775 780

Arg Arg Tyr Leu Glu Met Phe Tyr Ile Leu Lys Phe Arg Glu Leu Ala

785 790 795 800

Lys Ser Val Pro Leu Ala Ile Asp Asp Lys

805 810

<210> 20

<211> 3604

<212> DNA

<213> Nicotiana tabacum

<400> 20

atgtttacat ggctgaaact caatataaaa aacaagggta ggtgatcaaa aatcgttgga 60

tgcttaaaat cagtagacgt tttgctaaat gagcgaccaa tgttattgaa aacgttcatg 120

ttttcaaccc ttttggcata catttgagca ttgcccaaga ttttggataa gtagatgcag 180

tgcttataat tttaaagcat tgtatcctgc cttgtttttc attgtcaaaa ttaattaact 240

tacaagtatt tctataagtt gcttcataaa ttagaagtaa atctggattg tgtaatgtta 300

ttcgcctcgt aaatactgaa agctgcttga acaagtgaaa aaacacagac aaacgtaaca 360

ttctccatgg attgatgaga cttgtaaaat acatatatag aaatttggct tgtaaccaca 420

tgtatattat gccatatgga tgtgacattg atgtgactag acctaaatgt tttgtttcca 480

tgtccactgg agttttacgt atagttaaga ggagaaaaga ctgaggaata ctaatgtatg 540

atggtacccc tttgcttctt gacctggata cccagtgttc ctattcatgc ctatactttg 600

gtccttgatt tcactctccc ttttctaact tgagctgcat caaagaaatt tccactgtaa 660

aaaaataaat aatgctcacc atatctctgc aacattgcaa acatgtatcc catatgattg 720

atattggtgc gacatggcaa acatgtatcc tatttgatga tcaatcaaat ttatttttcc 780

cctgtcaaaa tgacctcagt gtgtaattcc ctatgtattt gatagcattg taactcgtgt 840

catgattcat gaatagggta ctagaattgc atggttgaca aatattaact ggtcgattga 900

tgatccacaa aacatgcact tactgactaa aatgtgatgg gacagattta tttttgtttg 960

tgattaacac agtacttaac cctatactta atacaatttg gcctagctac aatcttttct 1020

tcagtgcaaa ttccttgtta cacgaccaat attgtctttc tgagttctat tctgttgtta 1080

cttacacttt tattattcga ataagacatt agattgcttg catgcaaatt gatagccact 1140

tgtttcagga agaatactgt tggtcagtac gagagccata ctgcattcac cctcccagga 1200

ctatatcgcg tcgttcatgg cattgatgtt ttcgatccca aattcaatat agtgtctcct 1260

ggagctgaca tgacaattta cttcccatat tctgacaagg aaaaaagact aacgtctttg 1320

catggctcga ttgagaagtt gttatttgat cctgcgcaga atgaagagca tatgtaagtg 1380

acatccatt gtacttattt taatttggaa tagatgacat acttatttgc atgaatataa 1440

actgacaacc cagagatttc ctacattaga aaaggagggt ctgatatgat tttctacaaa 1500

taaattccca gtgatattgt tcaaaaagtc ctggatactt tattatgaga gaaccagga 1560

tagatggcac tagaatccct taatcttgag aagtcgccac ttatcgctcc caacactttc 1620

tgagaccctc aagtaactac tattattgtt tgatatcttg gagaagctat aagaatcttt 1680

ttctccttat tgtaattttt tttacgtgac tttaaactta acttccaagc tccttctgat 1740

aaaatgcaaa aactgtctgt attcactgtc ttggtttatt aacaattgat ccaatcaaat 1800

gcatatggaa catctttctt tttgtttctt caaaagttcg tttgaggata aggagtagaa 1860

tctgagaaga tagactagta ggtaacctta ggggcggatg tagaaatcaa cgtatgggtt 1920

cagctttgtt gcagaccctg tatatgcatt aaaaaaatca ctaaataagt aaataattga 1980

ttttgaaccc agtaaatcaa aatgagttgt agtagaatcc tgaactcgaa ccgataaagt 2040

tggatccact acggggtaaa ctctaccttg agaagtgttt atatatgtcc ctaattattt 2100

cttttctgtt tcctttctat tttaattttt taagttcctt tttagatggt tttatttttt 2160

gacaagtggt aagttgttag tattccaaat taaatgccat tgccataact atatacattt 2220

ataaagattg attgacccta gtttctcatt cctaagatcc aaataaggca ataaacaata 2280

tgtcttagta cttgaacctg cttctggtgg tcaacacttg atcgcgtagt tagttataga 2340

tgactgtaaa aaccttaatc attttaatgg ttttgtcaaa gaacaaatat cggacatatt 2400

atagcgaatg gactattgta cttttcttct gattggtcat tttattgtga tccgtaagtt 2460

ggctgagact gatgtcatat ctttgcttac agaggtaatc tgaatgataa atcaaaaccc 2520

ataatttttt caatggcaag gctagaccat gttaagaaca ttacgggact agttgagtgc 2580

tatgctaaaa atgccacatt gagggaattg gctaaccttg ttgttgtagc tggatacaac 2640

gatgtaaaga aatccagtga tagagaagaa atagcagaaa ttgagaagat gcatgctctt 2700

attaagggagc ataaattgga tgggcaattc agatggatag cagcccaaac aaaccgggca 2760

cgtaatggtg agctctatcg ctatatagct gacaagagag gtatatttgt tcaggtacgc 2820

tgtttgtatt gtatttgtcc acattccttt ttttgcaccg aaagaaaggt tgttattgtg 2880

acaaatatgt ttgttttagc ctgcatttta tgaagcattt ggactcacgg tggttgaagc 2940

tatgacttgt ggtcttccaa catttgcaac ttgccatggt ggtccgaacg agatcattga 3000

acacggtgta tctgggttcc atattgatcc ttatcatccc gataaagctg ctgaactcat 3060

ggcagaattc tttcaacgct gcaaacaaga tcctactcac tgggaaaaaa tatctgcatc 3120

tggtctccga aggattcttg agaggtttgt agttgtgtac atatatagaa gattaaagat 3180

tgttcccttg atattatttg aatgaaaaat aacagtaaca tctctttttg aacatcgctc 3240

aagttcttgt gttaaataat tgttaggtat acgtggaaaa tttactccga gaggctgatg 3300

actttgtctg gtgtatatgg tttctggaag cttgtttcaa aacttgagag gcgcgaaact 3360

agacgatacc ttgagatgtt ctacattctc aaattccgcg agttggtgag tgcctttttg 3420

ctcattttca gttacaatca actatatatg tggtttaaat acgtattaag cataaacatg 3480

tccgtgattg cggctgtcga aaatgctatg gacatatcct gagctaagga gttttcaaga 3540

gaattgattt ggcttactct gtttacaggc aaaatctgtt cctctggcaa ttgatgacaa 3600

GTGA 3604

<210> 21

<211> 335

<212> PROTEIN

<213> Nicotiana tabacum

<400> 21

Met Phe Thr Trp Leu Lys Leu Asn Ile Lys Asn Lys Gly Arg Lys Asn

1 5 10 15

Thr Val Gly Gln Tyr Glu Ser His Thr Ala Phe Thr Leu Pro Gly Leu

20 25 30

Tyr Arg Val Val His Gly Ile Asp Val Phe Asp Pro Lys Phe Asn Ile

35 40 45

Val Ser Pro Gly Ala Asp Met Thr Ile Tyr Phe Pro Tyr Ser Asp Lys

50 55 60

Glu Lys Arg Leu Thr Ser Leu His Gly Ser Ile Glu Lys Leu Leu Phe

65 70 75 80

Asp Pro Ala Gln Asn Glu Glu His Ile Gly Asn Leu Asn Asp Lys Ser

85 90 95

Lys Pro Ile Ile Phe Ser Met Ala Arg Leu Asp His Val Lys Asn Ile

100 105 110

Thr Gly Leu Val Glu Cys Tyr Ala Lys Asn Ala Thr Leu Arg Glu Leu

115 120 125

Ala Asn Leu Val Val Val Ala Gly Tyr Asn Asp Val Lys Lys Ser Ser

130 135 140

Asp Arg Glu Glu Ile Ala Glu Ile Glu Lys Met His Ala Leu Ile Lys

145 150 155 160

Glu His Lys Leu Asp Gly Gln Phe Arg Trp Ile Ala Ala Gln Thr Asn

165 170 175

Arg Ala Arg Asn Gly Glu Leu Tyr Arg Tyr Ile Ala Asp Lys Arg Gly

180 185 190

Ile Phe Val Gln Pro Ala Phe Tyr Glu Ala Phe Gly Leu Thr Val Val

195 200 205

Glu Ala Met Thr Cys Gly Leu Pro Thr Phe Ala Thr Cys His Gly Gly

210 215 220

Pro Asn Glu Ile Ile Glu His Gly Val Ser Gly Phe His Ile Asp Pro

225 230 235 240

Tyr His Pro Asp Lys Ala Ala Glu Leu Met Ala Glu Phe Phe Gln Arg

245 250 255

Cys Lys Gln Asp Pro Thr His Trp Glu Lys Ile Ser Ala Ser Gly Leu

260 265 270

Arg Arg Ile Leu Glu Arg Tyr Thr Trp Lys Ile Tyr Ser Glu Arg Leu

275 280 285

Met Thr Leu Ser Gly Val Tyr Gly Phe Trp Lys Leu Val Ser Lys Leu

290 295 300

Glu Arg Arg Glu Thr Arg Arg Tyr Leu Glu Met Phe Tyr Ile Leu Lys

305 310 315 320

Phe Arg Glu Leu Ala Lys Ser Val Pro Leu Ala Ile Asp Asp Lys

325 330 335

<210> 22

<211> 4030

<212> DNA

<213> Nicotiana tabacum

<400> 22

atggcggaac gtgtgctgac tcgtgttcat agccttcgtg aacgtcttga tgctactttg 60

gctgctcatc gcaatgagat tttgctgttt ctttcaaggt atagtcttag cagattgttc 120

tttgatttag ttgttattgc cagttctaat gtatgggctt atatataaac aaagtgttga 180

agtatgcaac catataaact gacagcttaa aatgcttgag agaacacact tttatttatt 240

taattatgcc ttcagcacaa gaagtggaac ttgacgcaat ggaaccatag gtcacgggtt 300

caagtcttgg aacagcctgc aatctaaggc tgcgtgtagt agaccctagt ggtccggccc 360

ttccacatat ctcgcttagt gtaccgggcc cattgagtac gggttcggcc gaacccagtc 420

gctttggtcc aatccatata tttgtcttaa aaatatattg aatatataca aattgttaat 480

ttagtttaaa tatgtgtatc atgggttatt catgctggtt ttggctgttg caggattgaa 540

agccatggaa aagggatact gaaacctcac cagttgctgg ctgaatttga ttcaattcac 600

aaagaagaca aaaacaaact gaatgatcat gcttttgaag aagtcctgaa atccactcag 660

gtatttgtgg ttttagtgtt aggtgatgga tagcatttat tgttttacta agatcacata 720

tgtgtcagtt tgtggctagt atttaaaatc tggtgtattt tgtcatacta ggaagcaatt 780

gttttgtccc cttgggttgc gcttgccatt cgtctgaggc ctggtgtgtg ggaatacgtt 840

cgtgtgaatg tcaacgctct tgttgttgag gagcttaccg tgcctgagta tttgcaattc 900

aaggaagaac ttgttaatgg aacgtaagtt ttaggttcga atttgttgat ttgttagata 960

acatgttctg aactttttga ttaaagttgt gtttttgact gatgcagctc gcacgataac 1020

tttgttcttg agttggattt tgagcccttc actgcatcat ttccaaaacc aaccctcacc 1080

aaatcaattg gaaatggagt tgaattcctt aaccgacacc tctctgccaa aatgttccat 1140

gacaaggaaa gcatgacccc tcttctcgag tttcttcgag ttcaccacta caagggcaag 1200

gtaaacttgt ttttcctgtt tgtctatgaa tttagtttag ttgttttgct ccgcgaaaat 1260

ttcagtggaa actgatttat gcaaccactg agtgattaat atgttcaaac ttaccgactt 1320

ctggttttct gtgtagacaa tgatgctgaa tgacagaatt caggacttaa atactctcca 1380

aaatgtccta aggaaagctg aggaatacct cactaccctt tcccctgaaa cttcatactc 1440

ggcatttgag cacaagttcc aagaaattgg cttggagagg ggttggggtg acactgcgga 1500

gcgtgttcta gagatgatct gcatgctcct ggatctcctc gaggctcctg actcgtgcac 1560

gcttgagaag ttccttggta gaattccaat ggtttttaat gtggtcatac tttcacccca 1620

tggttatttc gcccaggaaa atgtcttggg ttaccccgac actggtggcc aggtgcactg 1680

cttatctgtg ttcggtctta ttatctcttt aaaccctact gccacaagtg ctgagatgaa 1740

cctcctttaa tttgcaggtt gtctatattt tggatcaagt tcctgctttg gagcgtgaga 1800

tgctcaagcg cataaaggag caaggacttg acatcaaacc gcgtattctt attgttcgta 1860

ttcccagtaa ttgtgtttaa acttatgatt atgcaggatt ttatctgttc taatacagca 1920

ctcttgctta aattctcagg ttactcggct gctgcctgat gcggttggta ccacttgtgg 1980

tcagaggctt gagaaagtgt ttggaacaga gcactcacac attcttaggg tcccctttag 2040

gaccgagaag ggcattgttc gcaaatggat ctctcgcttt gaagtctggc catacatgga 2100

gacattcact gaggtgaagc aagctttctc tattcatttt tcaatcttcc aattggtttt 2160

ggcagcaatt ttctgcttgc tttgacttcc gctaaaactt cggattttat tgcattagga 2220

tgtggcgaaa gaaattgctg cagaattgca ggctaagcca gatcttatca ttggcaatta 2280

tagtgaggc aaccttgctg cctccttgtt ggctcacaaa ttaggtgtaa cacaggtcgg 2340

caatgtttgt gacatgtaat ttcatctttg catttccttt cgtttgcaac taaaagattt 2400

aagagttctc tctctctttt ttttttccgt ctactttgcc ttatgcagtg cacgatagct 2460

catgctttgg agaaaacaaa atatcctgat tctgatatct acttgaagaa atttgatgaa 2520

aaataccatt tctcagccca gtttactgcc gatcttattg caatgaatca caccgatttc 2580

atcatcacca gcactttcca ggagatagcg ggaaggtatt tttacatcag tttcccactc 2640

tgattaaatt acaatgtatt tccctatatg attaaatact gtgtttgatc ctaaatcatt 2700

tctaaatttt ccagcaagga cactgttgga cagtacgaga gccacatggc gttcacaatg 2760

cctggactgt atagagttgt tcacggcatt gatgtgtttg accccaaatt taacattgtg 2820

tcaccaggag ctgatatgaa tctctatttc ccatactacg agaaggaaaa gagattgaca 2880

gcatatcacc ctgaaattga ggagctgctg tttagtgatg ttgagaatga cgaacacatg 2940

tatgttacta aactagcaat cctgctgcaa aattatggct aattatgtaa acaagtttgt 3000

actgaataga tttgttattc gatcaggtgt gtgctgaaga acaggaataa gcctatcata 3060

ttcactatgg ctagattgga tcgagtgaag aacttaactg gacttgtcga gctgtacgcc 3120

aagaacccac ggctaaggga gttggttaac cttgtcgtgg ttggaggaga ccgaaggaaa 3180

gaatccaaag acttggaaga acaggcagag atgaagaaga tgtacgaact tataaagact 3240

cacaatttga acggccaatt ccgatggatt tcttcccaga tgaaccgcgt gaggaatggc 3300

gaactctaca ggtacattgc cgatactagg ggagctttcg tgcagcctgc attttacgag 3360

gcttttggtt tgactgttgt tgaggccatg acctgtggtt tgcctacatt tgcaactaat 3420

cacggtggtc cagctgagat catcgttcac gggaaatctg gtttccacat tgatccatac 3480

cacggggatc aggcagctga acttctcgct gatttctttg agaaatgtaa gaaagaacct 3540

tcgcactggg aagccatttc cgagggcggc cttaagcgta tacaggagaa gtaagcaaac 3600

tgctactctt ttcatttttg caaaacctac tatgatcatt attaagctca tttttgcaaa 3660

acctacttgc tgttgttatt gtttgttgct tccttttcac tgttctttga gctgaaggtc 3720

tatcagaaac agtctctcta ccttcacaag gtaggggtaa gatctgcgtg cacgttaccc 3780

tcctcaaact ctacttaatt gtgagattac actaggtttg ttgttgttga ttctttgcta 3840

attaattaaa aggtacacat ggcaaatata ctcggatcgg ttgttgacac tggctgctgt 3900

atatggattc tggaagcatg tttccaagct tgatcgtctt gaaattcgcc gttatcttga 3960

aatgttctat gctctcaaat tccgcaagct ggtgagtttc attgctttct gcactcctgc 4020

aattgtatag 4030

<210> 23

<211> 808

<212> PROTEIN

<213> Nicotiana tabacum

<400> 23

Met Ala Glu Arg Val Leu Thr Arg Val His Ser Leu Arg Glu Arg Leu

1 5 10 15

Asp Ala Thr Leu Ala Ala His Arg Asn Glu Ile Leu Leu Phe Leu Ser

20 25 30

Arg Ile Glu Ser His Gly Lys Gly Ile Leu Lys Pro His Gln Leu Leu

35 40 45

Ala Glu Phe Asp Ser Ile His Lys Glu Asp Lys Asn Lys Leu Asn Asp

50 55 60

His Ala Phe Glu Glu Val Leu Lys Ser Thr Gln Glu Ala Ile Val Leu

65 70 75 80

Ser Pro Trp Val Ala Leu Ala Ile Arg Leu Arg Pro Gly Val Trp Glu

85 90 95

Tyr Val Arg Val Asn Val Asn Ala Leu Val Val Glu Glu Leu Thr Val

100 105 110

Pro Glu Tyr Leu Gln Phe Lys Glu Glu Leu Val Asn Gly Thr Ser His

115 120 125

Asp Asn Phe Val Leu Glu Leu Asp Phe Glu Pro Phe Thr Ala Ser Phe

130 135 140

Pro Lys Pro Thr Leu Thr Lys Ser Ile Gly Asn Gly Val Glu Phe Leu

145 150 155 160

Asn Arg His Leu Ser Ala Lys Met Phe His Asp Lys Glu Ser Met Thr

165 170 175

Pro Leu Leu Glu Phe Leu Arg Val His His Tyr Lys Gly Lys Thr Met

180 185 190

Met Leu Asn Asp Arg Ile Gln Asp Leu Asn Thr Leu Gln Asn Val Leu

195 200 205

Arg Lys Ala Glu Glu Tyr Leu Thr Thr Leu Ser Pro Glu Thr Ser Tyr

210 215 220

Ser Ala Phe Glu His Lys Phe Gln Glu Ile Gly Leu Glu Arg Gly Trp

225 230 235 240

Gly Asp Thr Ala Glu Arg Val Leu Glu Met Ile Cys Met Leu Leu Asp

245 250 255

Leu Leu Glu Ala Pro Asp Ser Cys Thr Leu Glu Lys Phe Leu Gly Arg

260 265 270

Ile Pro Met Val Phe Asn Val Val Ile Leu Ser Pro His Gly Tyr Phe

275 280 285

Ala Gln Glu Asn Val Leu Gly Tyr Pro Asp Thr Gly Gly Gln Val Val

290 295 300

Tyr Ile Leu Asp Gln Val Pro Ala Leu Glu Arg Glu Met Leu Lys Arg

305 310 315 320

Ile Lys Glu Gln Gly Leu Asp Ile Lys Pro Arg Ile Leu Ile Val Thr

325 330 335

Arg Leu Leu Pro Asp Ala Val Gly Thr Thr Cys Gly Gln Arg Leu Glu

340 345 350

Lys Val Phe Gly Thr Glu His Ser His Ile Leu Arg Val Pro Phe Arg

355 360 365

Thr Glu Lys Gly Ile Val Arg Lys Trp Ile Ser Arg Phe Glu Val Trp

370 375 380

Pro Tyr Met Glu Thr Phe Thr Glu Asp Val Ala Lys Glu Ile Ala Ala

385 390 395 400

Glu Leu Gln Ala Lys Pro Asp Leu Ile Ile Gly Asn Tyr Ser Glu Gly

405 410 415

Asn Leu Ala Ala Ser Leu Leu Ala His Lys Leu Gly Val Thr Gln Cys

420 425 430

Thr Ile Ala His Ala Leu Glu Lys Thr Lys Tyr Pro Asp Ser Asp Ile

435 440 445

Tyr Leu Lys Lys Phe Asp Glu Lys Tyr His Phe Ser Ala Gln Phe Thr

450 455 460

Ala Asp Leu Ile Ala Met Asn His Thr Asp Phe Ile Ile Thr Ser Thr

465 470 475 480

Phe Gln Glu Ile Ala Gly Ser Lys Asp Thr Val Gly Gln Tyr Glu Ser

485 490 495

His Met Ala Phe Thr Met Pro Gly Leu Tyr Arg Val Val His Gly Ile

500 505 510

Asp Val Phe Asp Pro Lys Phe Asn Ile Val Ser Pro Gly Ala Asp Met

515 520 525

Asn Leu Tyr Phe Pro Tyr Tyr Glu Lys Glu Lys Arg Leu Thr Ala Tyr

530 535 540

His Pro Glu Ile Glu Glu Leu Leu Phe Ser Asp Val Glu Asn Asp Glu

545 550 555 560

His Met Cys Val Leu Lys Asn Arg Asn Lys Pro Ile Ile Phe Thr Met

565 570 575

Ala Arg Leu Asp Arg Val Lys Asn Leu Thr Gly Leu Val Glu Leu Tyr

580 585 590

Ala Lys Asn Pro Arg Leu Arg Glu Leu Val Asn Leu Val Val Val Gly

595 600 605

Gly Asp Arg Arg Lys Glu Ser Lys Asp Leu Glu Glu Gln Ala Glu Met

610 615 620

Lys Lys Met Tyr Glu Leu Ile Lys Thr His Asn Leu Asn Gly Gln Phe

625 630 635 640

Arg Trp Ile Ser Ser Gln Met Asn Arg Val Arg Asn Gly Glu Leu Tyr

645 650 655

Arg Tyr Ile Ala Asp Thr Arg Gly Ala Phe Val Gln Pro Ala Phe Tyr

660 665 670

Glu Ala Phe Gly Leu Thr Val Val Glu Ala Met Thr Cys Gly Leu Pro

675 680 685

Thr Phe Ala Thr Asn His Gly Gly Pro Ala Glu Ile Ile Val His Gly

690 695 700

Lys Ser Gly Phe His Ile Asp Pro Tyr His Gly Asp Gln Ala Ala Glu

705 710 715 720

Leu Leu Ala Asp Phe Phe Glu Lys Cys Lys Lys Glu Pro Ser His Trp

725 730 735

Glu Ala Ile Ser Glu Gly Gly Leu Lys Arg Ile Gln Glu Lys Tyr Thr

740 745 750

Trp Gln Ile Tyr Ser Asp Arg Leu Leu Thr Leu Ala Ala Val Tyr Gly

755 760 765

Phe Trp Lys His Val Ser Lys Leu Asp Arg Leu Glu Ile Arg Arg Tyr

770 775 780

Leu Glu Met Phe Tyr Ala Leu Lys Phe Arg Lys Leu Val Ser Phe Ile

785 790 795 800

Ala Phe Cys Thr Pro Ala Ile Val

805

<210> 24

<211> 4054

<212> DNA

<213> Nicotiana tabacum

<400> 24

atggccgaac gtgtgctaac tcgtgttcac agccttcgcg aacgtcttga tgctactttg 60

gctgctcatc gcaatgagat tttgctgttt ctttcaaggt atagtcttag cagattgttc 120

tttgatttag ttggtgttat ttgccagttc taatgtatgg actaatatat gaacaaagtg 180

cgaccatttc aactgacaac ttaaaatgtt tgagagaata cacgtttatt tacttaatta 240

tggcttgagc ataggaagtg tatcttggcg taactcgtaa agttgacctc atgtgacaag 300

gaggtcacgg tttcgagccg tggaaacagc ctcttgcaga aatgcaggta aggctgcgtg 360

caatagatcg cccttccacg gacccgcgca tagcgggaac ttagtgcacc ggttgggctg 420

tcctttttta tgtcttcagc acaaaaattt agtttaaaca tgtgtatcat ggattattca 480

tgctggtttt gccggttgca ggattgaaag ccacggaaaa gggatattga aacctcacca 540

gttgctggct gagtttgaat caattcacaa agaagacaaa aacaaactga atgatcatgc 600

ttttgaagaa gtcctgaaat ctactcaggt aatttgtggt tttagtgtta ggtgatggat 660

agcatttatt gtcttactaa gatcatatat gtgtcagttt gtggctagta tttgaaaagt 720

ctggtgtggt ttgtcatact aggaagcaat tgtcttgtcc ccttgggttg cgcttgccat 780

tcgtctgcgg cctggtgtgt gggaatatgt tcgtgtgaat gtcaatgcac ttattgtcga 840

ggagctgact gtgcctgaat atttgcaatt caaggaagaa cttgttaatg gaacgtaagt 900

tttaggttcg aaatgatgat ttgttaaata atatgttctg aactttttga ttaatgttgt 960

gttttcccct gatgcagctc gaacgataac tttgttcttg agctggattt tgagcccttc 1020

actgcatcat ttcccaaacc aaccctcacc aaatcaattg gaaatggagt tgaattcctc 1080

aaccgacacc tctctgccaa aatgttccat gacaaggaaa gcatgacccc tcttctcgag 1140

tttcttcgag ttcatcacta caagggcaag gtaaacttgt ttttcctgtt tgtctatgaa 1200

tttagtttct gaaagttgct ttgcttcgtg aattttttag tggcaactga tttatgattt 1260

tctgtgcaga caatgatgct gaatgacaga gttcaggact taaacactct ccaaaatgtc 1320

ctaaggaagg ctgaggaata tctcactacc ctttcccctg aaacttcata ctcggtattt 1380

gagcacaagt tccaagaaat tggcctagag aggggctggg gtgacaatgc tgagcgtgtt 1440

ctagagatga tctgcatgct cctggatctc ctcgaggctc cagactcatg cactcttgag 1500

aagttccttg gtagaattcc tatggttttt aatgtggtca ttctttcacc tcacggatat 1560

ttcgcccagg aaaatgtctt gggttacccc gatactggtg gccaggtgca ctgcttattt 1620

gtaacacctt acgcttttcc ctctgaaact tatttgcggc aagttctaag gtcctccttc 1680

cttaatttgc aggttgtcta tattttggat caagttccgg ccttggagcg tgagatgctc 1740

aagcgcataa aggagcaagg acttgatatc aaaccgcgta ttcttattgt tcgtatctcc 1800

aataattgcg tttaaactta tgattgtgca ggatttgatc tgttcaaatc taatgactga 1860

ttttcttttt tttttttttt tccctcaggt tactcggctg ctgcctgatg cggttggtac 1920

cacttgtggt cagcggcttg agaaagtgtt tggaacagag cattcacata ttcttagggt 1980

cccctttagg accgagaagg gcatcgttcg caaatggatc tctcgctttg aagtctggcc 2040

ttacatggag acattcactg aggtgaagca agctttctct attcattttt caatcttcca 2100

atctgttttg gcagcaattt ttcacttact aacactttgg ctttcgctaa aacttcggat 2160

tttattacat taggatgtgg caaaagaaat tgctgcagaa ctgcaggcaa agccagatct 2220

tataatcggc aactacagcg agggcaacct tgctgcctcc ttgttggctc acaagttagg 2280

tgtaactcag gtctgtaatg tttgtcacct gttatttcaa ctttgcattt cctttcattt 2340

gcaactagaa gttaagagtt ctctctcttt tatcttttcc gtctattttg ccttctgcag 2400

tgcaccatag ctcatgcgtt ggagaaaaca aaatatcctg attctgatat ctacttgaag 2460

aaatttgatg aaaaatacca tttctcagcc cagtttactg ccgatcttat tgcaatgaat 2520

cacaccgatt tcataatcac cagcactttc caggagatag cgggaaggta ttacatcaca 2580

atggatttcc gatatgatta aattagttaa tttaatccta cttcattgtg tttgatccta 2640

aaacttttct aaatttccca gcaaggacac tgttggacag tacgagagcc acatggcttt 2700

cacgatgcct ggattgtata gagttgttca cggcattgat gtgttcgatc ccaaattcaa 2760

cattgtgtca ccaggagctg atatgaatct ctatttcccc tacttcgaga aggaaaagcg 2820

attgacagca tatcaccctg aaattgagga gctgctgttt agcgatgttg agaatgacga 2880

acacatgtat gttactaaac tagcaatcct gctgcaaaat tgtggctaat tatgtaaaaa 2940

agtttttact gaatagattt gtgcttctat caggtgtgtg ctgaaggaca ggaataagcc 3000

aattatattc accatggcta gattggatcg agtgaagaac ttaactggac ttgtggagtt 3060

gtacgccaag aacccacggc taagggagtt ggttaacctt gtcgtggttg gtggagaccg 3120

aaggaaggaa tccaaagatt tggaagaaca ggcagagatg aagaagatgt atgaacttat 3180

aaagacgcac aatttaaacg gccaattccg atggatttct tccgatga accgcgtgag 3240

gaatggcgaa ctctacaggt acattgccga tactagggga gcttttgtgc agcctgcatt 3300

ttacgaggct tttggtttga ctgttgttga ggccatgacc tgtggtttgc ctacgtttgc 3360

aactaatcac ggtggtccag ctgagatcat cgttcacggg aagtctggtt ttcacattga 3420

tccataccac ggcgagcagg cagctgaact tctagctgat ttctttgaga gatgtaagaa 3480

agaaccttca cactgggaag ccatttccga gggcggcctt aagcgtatac aggagaagta 3540

agcaagctgc tactcttttc atttttgcaa aacctaccat gatcattatt aagctcattt 3600

ttgcaaaacc tacttgttat tctttgttgc ttccttttcc ctgttttttg agccgaggtt 3660

ttatcgaaaa catgctttct accttcacaa ggtaggggta aggtctgcgt ttgttattat 3720

tgttgttgtt gattctctgc gaattaatta aaaggtacac atggcaaatc tactcggatc 3780

ggttgttgac actggctgct gtttatggat tctggaagca tgtttccaaa cttgatcgtc 3840

ttgaaattcg tcgttatctt gaaatgttct atgctctaaa attccgcaaa ctggtgagtt 3900

tcactgcttt ctgcactctt ccaattgtta gttgagtgca ctcatttaaa ctgtagctaa 3960

agctgttgta aatcttcagt taagcagctg ctaatgaagt ttttatcttt tgtttttggt 4020

tcaggctgaa gctgtcccgt tggctgttga gtaa 4054

<210> 25

<211> 805

<212> PROTEIN

<213> Nicotiana tabacum

<400> 25

Met Ala Glu Arg Val Leu Thr Arg Val His Ser Leu Arg Glu Arg Leu

1 5 10 15

Asp Ala Thr Leu Ala Ala His Arg Asn Glu Ile Leu Leu Phe Leu Ser

20 25 30

Arg Ile Glu Ser His Gly Lys Gly Ile Leu Lys Pro His Gln Leu Leu

35 40 45

Ala Glu Phe Glu Ser Ile His Lys Glu Asp Lys Asn Lys Leu Asn Asp

50 55 60

His Ala Phe Glu Glu Val Leu Lys Ser Thr Gln Glu Ala Ile Val Leu

65 70 75 80

Ser Pro Trp Val Ala Leu Ala Ile Arg Leu Arg Pro Gly Val Trp Glu

85 90 95

Tyr Val Arg Val Asn Val Asn Ala Leu Ile Val Glu Glu Leu Thr Val

100 105 110

Pro Glu Tyr Leu Gln Phe Lys Glu Glu Leu Val Asn Gly Thr Ser Asn

115 120 125

Asp Asn Phe Val Leu Glu Leu Asp Phe Glu Pro Phe Thr Ala Ser Phe

130 135 140

Pro Lys Pro Thr Leu Thr Lys Ser Ile Gly Asn Gly Val Glu Phe Leu

145 150 155 160

Asn Arg His Leu Ser Ala Lys Met Phe His Asp Lys Glu Ser Met Thr

165 170 175

Pro Leu Leu Glu Phe Leu Arg Val His His Tyr Lys Gly Lys Thr Met

180 185 190

Met Leu Asn Asp Arg Val Gln Asp Leu Asn Thr Leu Gln Asn Val Leu

195 200 205

Arg Lys Ala Glu Glu Tyr Leu Thr Thr Leu Ser Pro Glu Thr Ser Tyr

210 215 220

Ser Val Phe Glu His Lys Phe Gln Glu Ile Gly Leu Glu Arg Gly Trp

225 230 235 240

Gly Asp Asn Ala Glu Arg Val Leu Glu Met Ile Cys Met Leu Leu Asp

245 250 255

Leu Leu Glu Ala Pro Asp Ser Cys Thr Leu Glu Lys Phe Leu Gly Arg

260 265 270

Ile Pro Met Val Phe Asn Val Val Ile Leu Ser Pro His Gly Tyr Phe

275 280 285

Ala Gln Glu Asn Val Leu Gly Tyr Pro Asp Thr Gly Gly Gln Val Val

290 295 300

Tyr Ile Leu Asp Gln Val Pro Ala Leu Glu Arg Glu Met Leu Lys Arg

305 310 315 320

Ile Lys Glu Gln Gly Leu Asp Ile Lys Pro Arg Ile Leu Ile Val Thr

325 330 335

Arg Leu Leu Pro Asp Ala Val Gly Thr Thr Cys Gly Gln Arg Leu Glu

340 345 350

Lys Val Phe Gly Thr Glu His Ser His Ile Leu Arg Val Pro Phe Arg

355 360 365

Thr Glu Lys Gly Ile Val Arg Lys Trp Ile Ser Arg Phe Glu Val Trp

370 375 380

Pro Tyr Met Glu Thr Phe Thr Glu Asp Val Ala Lys Glu Ile Ala Ala

385 390 395 400

Glu Leu Gln Ala Lys Pro Asp Leu Ile Ile Gly Asn Tyr Ser Glu Gly

405 410 415

Asn Leu Ala Ala Ser Leu Leu Ala His Lys Leu Gly Val Thr Gln Cys

420 425 430

Thr Ile Ala His Ala Leu Glu Lys Thr Lys Tyr Pro Asp Ser Asp Ile

435 440 445

Tyr Leu Lys Lys Phe Asp Glu Lys Tyr His Phe Ser Ala Gln Phe Thr

450 455 460

Ala Asp Leu Ile Ala Met Asn His Thr Asp Phe Ile Ile Thr Ser Thr

465 470 475 480

Phe Gln Glu Ile Ala Gly Ser Lys Asp Thr Val Gly Gln Tyr Glu Ser

485 490 495

His Met Ala Phe Thr Met Pro Gly Leu Tyr Arg Val Val His Gly Ile

500 505 510

Asp Val Phe Asp Pro Lys Phe Asn Ile Val Ser Pro Gly Ala Asp Met

515 520 525

Asn Leu Tyr Phe Pro Tyr Phe Glu Lys Glu Lys Arg Leu Thr Ala Tyr

530 535 540

His Pro Glu Ile Glu Glu Leu Leu Phe Ser Asp Val Glu Asn Asp Glu

545 550 555 560

His Met Cys Val Leu Lys Asp Arg Asn Lys Pro Ile Ile Phe Thr Met

565 570 575

Ala Arg Leu Asp Arg Val Lys Asn Leu Thr Gly Leu Val Glu Leu Tyr

580 585 590

Ala Lys Asn Pro Arg Leu Arg Glu Leu Val Asn Leu Val Val Val Gly

595 600 605

Gly Asp Arg Arg Lys Glu Ser Lys Asp Leu Glu Glu Gln Ala Glu Met

610 615 620

Lys Lys Met Tyr Glu Leu Ile Lys Thr His Asn Leu Asn Gly Gln Phe

625 630 635 640

Arg Trp Ile Ser Ser Gln Met Asn Arg Val Arg Asn Gly Glu Leu Tyr

645 650 655

Arg Tyr Ile Ala Asp Thr Arg Gly Ala Phe Val Gln Pro Ala Phe Tyr

660 665 670

Glu Ala Phe Gly Leu Thr Val Val Glu Ala Met Thr Cys Gly Leu Pro

675 680 685

Thr Phe Ala Thr Asn His Gly Gly Pro Ala Glu Ile Ile Val His Gly

690 695 700

Lys Ser Gly Phe His Ile Asp Pro Tyr His Gly Glu Gln Ala Ala Glu

705 710 715 720

Leu Leu Ala Asp Phe Phe Glu Arg Cys Lys Lys Glu Pro Ser His Trp

725 730 735

Glu Ala Ile Ser Glu Gly Gly Leu Lys Arg Ile Gln Glu Lys Tyr Thr

740 745 750

Trp Gln Ile Tyr Ser Asp Arg Leu Leu Thr Leu Ala Ala Val Tyr Gly

755 760 765

Phe Trp Lys His Val Ser Lys Leu Asp Arg Leu Glu Ile Arg Arg Tyr

770 775 780

Leu Glu Met Phe Tyr Ala Leu Lys Phe Arg Lys Leu Ala Glu Ala Val

785 790 795 800

About Leu Ala Val Glu

805

<210> 26

<211> 3752

<212> DNA

<213> Nicotiana tabacum

<400> 26

atggcctcaa cagttgctga tagcatgcct gatgctttga aacaaagccg gtatcatatg 60

aagagatgct tcgctaggtg aacacccttc ttttatgttt tttcccctct acgtgtttat 120

gtcaaatttc catgcataat gctaactact tttcttcttt ttgacttcaa aattggatgt 180

gaaaggttca ttgcaatggg aaggaggcta atgaagttga aacatttaac agaagaaata 240

gaagaaacta ttgaagacaa ggcagaaaga accaggattt tggagggttc acttggaaaa 300

attatgagtt ccacacaggt cagcaccatt taaccaactt agttgaacag gaaaaaaaga 360

aaaagcaaaa gagttattgc aaggcgtaac gattttcttt gaaattttca ggaggcagct 420

gttgttccac cttatgttgc ttttgcagta aggcacaatc ctggcttctg ggattatgtc 480

aaagttaacg ctgaaactct ctctgtggaa gctatttcag ccagggaata tctcaaattc 540

aaagagatga tctttgacga agactggtaa gtggaaaatt gtatcatttt aaagagaaac 600

aattttgtaa catacaagaa tagttttgat ggttgaatgt gcaagcaggg caaaggatga 660

taatgcactg gaagtagatt ttggtgcttt tgactactct aatcctcggt tagccctttc 720

ctcttctgtc ggaaatgggc tcaactttat ctcaaaagtt ctgtcttcaa agtttggtgg 780

aaagccagag gacgcccagc ctttgcttga ttacttacta gctcttaatc atcaaggaga 840

ggtatgaaaa tggactacct ttgtttctta aaggtattat ataatgatgc gcgttataaa 900

gttccttttt aaattgaaac tttgcagaat ctaatgatca atgagaatct gaatggtgtt 960

gctaagcttc aagcagcatt gatagtagct gaagtttttg tatcttcctt tcccaaagac 1020

acaccttata aagactttga gcataagtaa gcttctcata tgcttccatt gtcatatgca 1080

gtataccaat gacatgctac cgaaaagttg tttatgtttg tgacttgatt atgaaaactc 1140

taggctcaaa gaatggggct ttgataaagg gtggggtcac aatgcaggaa gagtaagaga 1200

gacaatgaga ctgctttccg agataatcca agcaccagat cccataaata tggagtcctt 1260

tttcagcaag cttcctacta cattcaacat tgttatcttc tccattcatg gttactttgg 1320

ccaagcagat gtccttggtc tgcccgatac tggaggccag gtctacatat acagcaattt 1380

atctcctttt gcctcatatt gcttattagc gacacttgca tcattgaaat cagactttta 1440

cttcacaggt tgtttatatt ctggatcaag taagggcttt agaggaggaa atgttacaaa 1500

gaatcaagca gcaagggcta aacgtgaagc ccaagattct tgtggtgagt tttgcaaaaa 1560

tatgcttaga caggttttga gattgatcgg agaagggatt aagatgatca agatctttgt 1620

ttcctgcttt catgatgtaa acaggtatct cgtctcatac cagatgctcg agggacaaca 1680

tgcaatcagg agatggaacc tattcttaac tcatcccatt ctcacatcct gagaattcca 1740

ttcaggactg agaaaggagt tcttcgccaa tgggtttctc ggtttgatat ctatccttac 1800

ttggagaact atgccaaggc aagtcttcta acaaaattac cacctattca tacactttat 1860

ttactttctt gaactaatcg tttggtttgt gacgtatatc attaggatgc ttctgctaag 1920

atacttgagc tcatggaagg taaaccagac ctcataattg ggaactacac tgatggaaat 1980

ttagtggcat ctctattggc caacaaactt ggagttactc aggttccgta gctgatcata 2040

tgatcatatt ttctacattg tttcttgata attaaatgga aatcttattg gatgataaca 2100

ttttagggaa ccattgctca tgcattagag aaaactaagt atgaagattc tgatgtgaag 2160

tggaagcagt ttgatcccaa gtaccacttt tcttgccaat ttactgccga tttattggca 2220

atgaatgctg ctgattttat cattaccagc acatatcaag aaatcgctgg aaggttagca 2280

ctgactctct cagtatattt ggcaacttaa tgaatttact gcagtggcca acactaaaag 2340

ctatcattcg tccttcagcg aaactaggcc tggacaatat gaaagtcaca cagcatttac 2400

catgccgggg ctttatagag ctgtttcagg catcaatgta tttgatccaa agttcaacat 2460

tgctgctcct ggggctgaac agtctaccta tttccctttc actgagaaac agaaacgatt 2520

cagcacattt cgtcctgcta ttaacgaatt actttacagt aatgaggaaa acaatgagca 2580

catgtaagtc taattgccca ttttcctaat ctaaccattg cttaaatcgt tctgttttta 2640

ccggatgtgt ggtacttatc agtaacattt ttttttggat cagtggattt cttgcagacc 2700

ggaaaaaacc aattatattt tcaatggcga gatttgatac agtgaagaac ctgtcaggct 2760

tgactgagtg gtatgggaag aataagaagt tgcggaactt ggtaaacctt gttattgttg 2820

ggggattctt cgatccatca aaatcaaaag accgggagga agcagctgaa atcaagaaga 2880

tgcatgaatt gattgagaaa taccagctca agggacaaat gagatggata gcagctcaaa 2940

ctgataaata tcgaaatagt gagctatacc gaactattgc tgacactaag ggagcttttg 3000

tccaaccggc tttatatgaa gcttttggac taaccgttat tgaagcaatg gattgtggat 3060

tgcctacgtt tgcaactaat caaggtggac ctgcagaaat cattgttgat ggggtttcag 3120

gtttccatat tgatccttac aatggggacg aatcaagcaa gaaaatagct gatttctttg 3180

agaagtgtaa ggttgattct aaatattgga acaggatatc tgagggaggt ctcaagcgca 3240

ttgaagaatg gtaacaaact agttccaagt ttaaaaaatg gaaaaaatgc ttatcatgtt 3300

atattttcgt ggttttaagt tctgcttcga tgcagttata cgtggaagat ttatgcaaac 3360

aaagtgttga atatgggatc aatctatgga ttttggagac aattcaatgt ggggcaaaag 3420

caggctaagc aaagatactt tgagatgttt tacaatcctc tcttcaggaa attggtaggt 3480

tgtatatgtt gaatacaatt tactaagatc ctcaaaatga ccaagaaata tacattgact 3540

atgctacttt tgtaatttca caggccaaaa gcgtgccgat cccacatgaa gagccattgc 3600

cacttgcaac atcagactct actcaatccc aagaattaaa actaccacta ccagttccag 3660

cagcagtagc taaagttctg ccattaacaa ggcatgcttt taacttaatt acttctctac 3720

ctagagtaac tggtaaagtg gatgtcaagt ga 3752

<210> 27

<211> 840

<212> PROTEIN

<213> Nicotiana tabacum

<400> 27

Met Ala Ser Thr Val Ala Asp Ser Met Pro Asp Ala Leu Lys Gln Ser

1 5 10 15

Arg Tyr His Met Lys Arg Cys Phe Ala Arg Phe Ile Ala Met Gly Arg

20 25 30

Arg Leu Met Lys Leu Lys His Leu Thr Glu Glu Ile Glu Glu Thr Ile

35 40 45

Glu Asp Lys Ala Glu Arg Thr Arg Ile Leu Glu Gly Ser Leu Gly Lys

50 55 60

Ile Met Ser Ser Thr Gln Glu Ala Ala Val Val Pro Pro Tyr Val Ala

65 70 75 80

Phe Ala Val Arg His Asn Pro Gly Phe Trp Asp Tyr Val Lys Val Asn

85 90 95

Ala Glu Thr Leu Ser Val Glu Ala Ile Ser Ala Arg Glu Tyr Leu Lys

100 105 110

Phe Lys Glu Met Ile Phe Asp Glu Asp Trp Ala Lys Asp Asp Asn Ala

115 120 125

Leu Glu Val Asp Phe Gly Ala Phe Asp Tyr Ser Asn Pro Arg Leu Ala

130 135 140

Leu Ser Ser Ser Val Gly Asn Gly Leu Asn Phe Ile Ser Lys Val Leu

145 150 155 160

Ser Ser Lys Phe Gly Gly Lys Pro Glu Asp Ala Gln Pro Leu Leu Asp

165 170 175

Tyr Leu Leu Ala Leu Asn His Gln Gly Glu Asn Leu Met Ile Asn Glu

180 185 190

Asn Leu Asn Gly Val Ala Lys Leu Gln Ala Ala Leu Ile Val Ala Glu

195 200 205

Val Phe Val Ser Ser Phe Pro Lys Asp Thr Pro Tyr Lys Asp Phe Glu

210 215 220

His Lys Leu Lys Glu Trp Gly Phe Asp Lys Gly Trp Gly His Asn Ala

225 230 235 240

Gly Arg Val Arg Glu Thr Met Arg Leu Leu Ser Glu Ile Ile Gln Ala

245 250 255

Pro Asp Pro Ile Asn Met Glu Ser Phe Phe Ser Lys Leu Pro Thr Thr

260 265 270

Phe Asn Ile Val Ile Phe Ser Ile His Gly Tyr Phe Gly Gln Ala Asp

275 280 285

Val Leu Gly Leu Pro Asp Thr Gly Gly Gln Val Val Tyr Ile Leu Asp

290 295 300

Gln Val Arg Ala Leu Glu Glu Glu Met Leu Gln Arg Ile Lys Gln Gln

305 310 315 320

Gly Leu Asn Val Lys Pro Lys Ile Leu Val Val Ser Arg Leu Ile Pro

325 330 335

Asp Ala Arg Gly Thr Thr Cys Asn Gln Glu Met Glu Pro Ile Leu Asn

340 345 350

Ser Ser His Ser His Ile Leu Arg Ile Pro Phe Arg Thr Glu Lys Gly

355 360 365

Val Leu Arg Gln Trp Asp Ala Ser Ala Lys Ile Leu Glu Leu Met Glu

370 375 380

Gly Lys Pro Asp Leu Ile Ile Gly Asn Tyr Thr Asp Gly Asn Leu Val

385 390 395 400

Ala Ser Leu Leu Ala Asn Lys Leu Gly Val Thr Gln Gly Thr Ile Ala

405 410 415

His Ala Leu Glu Lys Thr Lys Tyr Glu Asp Ser Asp Val Lys Trp Lys

420 425 430

Gln Phe Asp Pro Lys Tyr His Phe Ser Cys Gln Phe Thr Ala Asp Leu

435 440 445

Leu Ala Met Asn Ala Ala Asp Phe Ile Ile Thr Ser Thr Tyr Gln Glu

450 455 460

Ile Ala Gly Ser Glu Thr Arg Pro Gly Gln Tyr Glu Ser His Thr Ala

465 470 475 480

Phe Thr Met Pro Gly Leu Tyr Arg Ala Val Ser Gly Ile Asn Val Phe

485 490 495

Asp Pro Lys Phe Asn Ile Ala Ala Pro Gly Ala Glu Gln Ser Thr Tyr

500 505 510

Phe Pro Phe Thr Glu Lys Gln Lys Arg Phe Ser Thr Phe Arg Pro Ala

515 520 525

Ile Asn Glu Leu Leu Tyr Ser Asn Glu Glu Asn Asn Glu His Ile Gly

530 535 540

Phe Leu Ala Asp Arg Lys Lys Pro Ile Ile Phe Ser Met Ala Arg Phe

545 550 555 560

Asp Thr Val Lys Asn Leu Ser Gly Leu Thr Glu Trp Tyr Gly Lys Asn

565 570 575

Lys Lys Leu Arg Asn Leu Val Asn Leu Val Ile Val Gly Gly Phe Phe

580 585 590

Asp Pro Ser Lys Ser Lys Asp Arg Glu Glu Ala Ala Glu Ile Lys Lys

595 600 605

Met His Glu Leu Ile Glu Lys Tyr Gln Leu Lys Gly Gln Met Arg Trp

610 615 620

Ile Ala Ala Gln Thr Asp Lys Tyr Arg Asn Ser Glu Leu Tyr Arg Thr

625 630 635 640

Ile Ala Asp Thr Lys Gly Ala Phe Val Gln Pro Ala Leu Tyr Glu Ala

645 650 655

Phe Gly Leu Thr Val Ile Glu Ala Met Asp Cys Gly Leu Pro Thr Phe

660 665 670

Ala Thr Asn Gln Gly Gly Pro Ala Glu Ile Ile Val Asp Gly Val Ser

675 680 685

Gly Phe His Ile Asp Pro Tyr Asn Gly Asp Glu Ser Ser Lys Lys Ile

690 695 700

Ala Asp Phe Phe Glu Lys Cys Lys Val Asp Ser Lys Tyr Trp Asn Arg

705 710 715 720

Ile Ser Glu Gly Gly Leu Lys Arg Ile Glu Glu Cys Tyr Thr Trp Lys

725 730 735

Ile Tyr Ala Asn Lys Val Leu Asn Met Gly Ser Ile Tyr Gly Phe Trp

740 745 750

Arg Gln Phe Asn Val Gly Gln Lys Gln Ala Lys Gln Arg Tyr Phe Glu

755 760 765

Met Phe Tyr Asn Pro Leu Phe Arg Lys Leu Ala Lys Ser Val Pro Ile

770 775 780

Pro His Glu Glu Pro Leu Pro Leu Ala Thr Ser Asp Ser Thr Gln Ser

785 790 795 800

Gln Glu Leu Lys Leu Pro Leu Pro Val Pro Ala Ala Val Ala Lys Val

805 810 815

Leu Pro Leu Thr Arg His Ala Phe Asn Leu Ile Thr Ser Leu Pro Arg

820 825 830

Val Thr Gly Lys Val Asp Val Lys

835 840

<210> 28

<211> 3268

<212> DNA

<213> Nicotiana tabacum

<400> 28

atggcctcaa ctgttgctgg tagcatgcct gatgctttga aacaaagccg atatcatatg 60

aagagatgct tcgctaggtg aacacccttc ttgttctttt tgttttttcc ctctaccatt 120

tatgtcaaat ttcaatgcat aatgctaact actttttttc tttttgactt caaaattgga 180

cgtgaaaggt tcattgcaat gggaaggagg ttgatgaagc tgaaacattt aacagaagaa 240

atagaaaaaa ctattgaaga caaggcagaa agaaccaaga ttttggaggg ttcacttgga 300

aaaattatga gttccacaca ggtcagcacc atttaaccaa cttaattgaa taggaagaaa 360

aaaaaaagca aaagagttat tgcaaggcgt aacgatttcc tttgaaattt tcaggaggca 420

gctgttgtcc caccttatgt tgcttttgca gtaaggcaca atcctggctt ctgggattat 480

gtcaaagttg acgctgaaac tctctctgtg gaagctattt cagccaggga ctatctcaaa 540

ttcaaagaga tgatctttga tgaagattgg taactggaag attgtatcat tttaaagaaa 600

caatttttta atattcaaga ttagttttga tggttgaatg tgcaagcagg gcaaaggatg 660

aaaatgcact cgaagtagat tttggtgctt ttgactactc taatcatcgg ttagcccttt 720

cctcttctgt cggaaatggg ctaaacttca tctcgaaagt tttgtcttca aagtttggtg 780

gaaaggcaga agatgcccag cctttgcttg attacttact agctcttaat catcaaggag 840

aggtatggaa atggactacc ttcctttctt aaggaattat ataatgatgt atgttataaa 900

gatccttttt aaacattgac actttgcaga atctaatgat caatgagaat ctgaatggcg 960

tctctaagct tcaagcagca ttgatagtag ctgaagtttt tgtatcttcc tttcccaaag 1020

acacacctta taaagacttt gagcataagt aagcttttca aacgcttctg ttatcatatg 1080

caatatacca agaatatgtt gccttttgaa aagttgttta tgtttatgac ttgataatga 1140

aaatactagg ctcaaagaat ggggctttga gaaagggtgg ggtcacaatg caggaagagt 1200

aagagagaca atgagactgc tttccgagat aatccaagcg ccagatccca taaatatgga 1260

gtcctttttc agcaggcttc ctactacatt caacattgtt atcttctcca ttcatggtta 1320

ctttggccaa gcagatgtcc ttggtttgcc cgatactgga ggccaggttt acatacacag 1380

caatttatct ccttttgcct catatttact tattagcgac acttgcatta ttgaaatcac 1440

atttgtattt aacaggttgt ttatattctg gatcaagtaa gagccttaga ggaggaaatg 1500

ttacaaagaa tcaagcagca agggttaaat gtgaagccca agattcttgt ggtgagttat 1560

gcaaaaatat gcgtagccaa ggttttgaaa ttgttcagag gggattaaga tgatcgagat 1620

atttgtttcc ttcttccatt gatgtgtaca ggtcactcgt ctcattccag atgctcgagg 1680

gactacatgc aatcaggaga tggaacctat acttaactcg tcccattctc acatcctgag 1740

aattccatc aggacagaga aaggagttct tcgccaatgg gtttctcggt ttgatatcta 1800

tccttacttg gagaactatg ccaaggcaag tctcctacca aaattaccac ctattcatac 1860

actttattca gttttttgag ctaatcattc tcatttgtca cgtatgtgat taggatgctt 1920

ctgctaagat acttgagctc atggaaggta aaccagacct cattattggg aactacactg 1980

atggaaattt agtggcatct ctattggcca acaaacttgg agttactcag gttctacagc 2040

tgatcattta tctgatcaga ttttctacat tgttttcttg ataattaaac ggaaatctta 2100

tgagattgta acattttagg gaaccattgc tcatgcatta gagaaaacca agtatgaaga 2160

ttctgatgtc aagtggaagc agtttgattc caagtaccac ttttcttgcc aattcactgc 2220

cgatttattg gcaatgaatg ctgctgattt tatcattacc agcacatatc aagaaatcgc 2280

aggaaggtta gcactgactc tctcagtata tttggcaact taatgaatgt actgcttgtg 2340

gccaacacta aaagctatta ctcgtccttc agcgaaacta ggcctggaca atatgaaagt 2400

cacacagcat ttaccatgcc ggggctttat agagctgttt caggcatcaa tgtatttgat 2460

ccaaagttca acattgctgc tcctggggct gaacagtctg cctatttccc cttcactgag 2520

aaacagaaac gattcagcgc gtttcgtcct gctattgagg aactacttta cagtaatgag 2580

caaaacaacg agcacatgta agtctaattg ccccattttc ctaatctaac cattgcttaa 2640

atgttctgtt tttacttgat atgtggtact tatcagtgat attttttatt ggaacagtgg 2700

atttcttgca gaccgtaaaa aaccaattat attttcaatg gcaagatttg atacggtgaa 2760

gaacttgtca ggcttgactg agtggtatgg gaagaataag aagttgcgga acttggttaa 2820

cctcgttatc gttgggggat tcttcgatcc atcaaaatca aaagaccggg aggaagcagc 2880

tgaaatcaag aagatgcatg aattgattga gaaatacaag ctcaagggac aaatgagatg 2940

gatagcagct caaactgata aatatcaaaa cagtgagcta tatcgaacta ttgctgacac 3000

taaaggagct ttcgtccaac cggctttata tgaagctttt ggactaactg ttattgaagc 3060

aatgaattgt ggactgccta catttgctac taatcaaggc ggacctgcag aaatcattgt 3120

tgatggggtt tcaggcttcc atattgatcc ttacaatggg gatgaatcga gcaagaaaat 3180

agctgatttc tttgagaagt gtaaggttga ttctaaatat tggaacaaga tatgtggagg 3240

aggtctcaag cgcattgaag aatggtaa 3268

<210> 29

<211> 732

<212> PROTEIN

<213> Nicotiana tabacum

<400> 29

Met Ala Ser Thr Val Ala Gly Ser Met Pro Asp Ala Leu Lys Gln Ser

1 5 10 15

Arg Tyr His Met Lys Arg Cys Phe Ala Arg Phe Ile Ala Met Gly Arg

20 25 30

Arg Leu Met Lys Leu Lys His Leu Thr Glu Glu Ile Glu Lys Thr Ile

35 40 45

Glu Asp Lys Ala Glu Arg Thr Lys Ile Leu Glu Gly Ser Leu Gly Lys

50 55 60

Ile Met Ser Ser Thr Gln Glu Ala Ala Val Val Pro Pro Tyr Val Ala

65 70 75 80

Phe Ala Val Arg His Asn Pro Gly Phe Trp Asp Tyr Val Lys Val Asp

85 90 95

Ala Glu Thr Leu Ser Val Glu Ala Ile Ser Ala Arg Asp Tyr Leu Lys

100 105 110

Phe Lys Glu Met Ile Phe Asp Glu Asp Trp Ala Lys Asp Glu Asn Ala

115 120 125

Leu Glu Val Asp Phe Gly Ala Phe Asp Tyr Ser Asn His Arg Leu Ala

130 135 140

Leu Ser Ser Ser Val Gly Asn Gly Leu Asn Phe Ile Ser Lys Val Leu

145 150 155 160

Ser Ser Lys Phe Gly Gly Lys Ala Glu Asp Ala Gln Pro Leu Leu Asp

165 170 175

Tyr Leu Leu Ala Leu Asn His Gln Gly Glu Asn Leu Met Ile Asn Glu

180 185 190

Asn Leu Asn Gly Val Ser Lys Leu Gln Ala Ala Leu Ile Val Ala Glu

195 200 205

Val Phe Val Ser Ser Phe Pro Lys Asp Thr Pro Tyr Lys Asp Phe Glu

210 215 220

His Lys Leu Lys Glu Trp Gly Phe Glu Lys Gly Trp Gly His Asn Ala

225 230 235 240

Gly Arg Val Arg Glu Thr Met Arg Leu Leu Ser Glu Ile Ile Gln Ala

245 250 255

Pro Asp Pro Ile Asn Met Glu Ser Phe Phe Ser Arg Leu Pro Thr Thr

260 265 270

Phe Asn Ile Val Ile Phe Ser Ile His Gly Tyr Phe Gly Gln Ala Asp

275 280 285

Val Leu Gly Leu Pro Asp Thr Gly Gly Gln Val Val Tyr Ile Leu Asp

290 295 300

Gln Val Arg Ala Leu Glu Glu Glu Met Leu Gln Arg Ile Lys Gln Gln

305 310 315 320

Gly Leu Asn Val Lys Pro Lys Ile Leu Val Val Thr Arg Leu Ile Pro

325 330 335

Asp Ala Arg Gly Thr Thr Cys Asn Gln Glu Met Glu Pro Ile Leu Asn

340 345 350

Ser Ser His Ser His Ile Leu Arg Ile Pro Phe Arg Thr Glu Lys Gly

355 360 365

Val Leu Arg Gln Trp Asp Ala Ser Ala Lys Ile Leu Glu Leu Met Glu

370 375 380

Gly Lys Pro Asp Leu Ile Ile Gly Asn Tyr Thr Asp Gly Asn Leu Val

385 390 395 400

Ala Ser Leu Leu Ala Asn Lys Leu Gly Val Thr Gln Gly Thr Ile Ala

405 410 415

His Ala Leu Glu Lys Thr Lys Tyr Glu Asp Ser Asp Val Lys Trp Lys

420 425 430

Gln Phe Asp Ser Lys Tyr His Phe Ser Cys Gln Phe Thr Ala Asp Leu

435 440 445

Leu Ala Met Asn Ala Ala Asp Phe Ile Ile Thr Ser Thr Tyr Gln Glu

450 455 460

Ile Ala Gly Ser Glu Thr Arg Pro Gly Gln Tyr Glu Ser His Thr Ala

465 470 475 480

Phe Thr Met Pro Gly Leu Tyr Arg Ala Val Ser Gly Ile Asn Val Phe

485 490 495

Asp Pro Lys Phe Asn Ile Ala Ala Pro Gly Ala Glu Gln Ser Ala Tyr

500 505 510

Phe Pro Phe Thr Glu Lys Gln Lys Arg Phe Ser Ala Phe Arg Pro Ala

515 520 525

Ile Glu Glu Leu Leu Tyr Ser Asn Glu Gln Asn Asn Glu His Ile Gly

530 535 540

Phe Leu Ala Asp Arg Lys Lys Pro Ile Ile Phe Ser Met Ala Arg Phe

545 550 555 560

Asp Thr Val Lys Asn Leu Ser Gly Leu Thr Glu Trp Tyr Gly Lys Asn

565 570 575

Lys Lys Leu Arg Asn Leu Val Asn Leu Val Ile Val Gly Gly Phe Phe

580 585 590

Asp Pro Ser Lys Ser Lys Asp Arg Glu Glu Ala Ala Glu Ile Lys Lys

595 600 605

Met His Glu Leu Ile Glu Lys Tyr Lys Leu Lys Gly Gln Met Arg Trp

610 615 620

Ile Ala Ala Gln Thr Asp Lys Tyr Gln Asn Ser Glu Leu Tyr Arg Thr

625 630 635 640

Ile Ala Asp Thr Lys Gly Ala Phe Val Gln Pro Ala Leu Tyr Glu Ala

645 650 655

Phe Gly Leu Thr Val Ile Glu Ala Met Asn Cys Gly Leu Pro Thr Phe

660 665 670

Ala Thr Asn Gln Gly Gly Pro Ala Glu Ile Ile Val Asp Gly Val Ser

675 680 685

Gly Phe His Ile Asp Pro Tyr Asn Gly Asp Glu Ser Ser Lys Lys Ile

690 695 700

Ala Asp Phe Phe Glu Lys Cys Lys Val Asp Ser Lys Tyr Trp Asn Lys

705 710 715 720

Ile Cys Gly Gly Gly Leu Lys Arg Ile Glu Glu Trp

725 730

<210> 30

<211> 3937

<212> DNA

<213> Nicotiana tabacum

<400> 30

atggctactg caccagccct aaatagatca gagtccatag ctgatagcat gccagaggcc 60

ttaaggcaaa gccggtacca catgaagaaa tgttttgcca agtacataga gcaaggaaag 120

aggatgatga aacttcataa cttgatggat gagttggaga aagtaattga tgatcctgct 180

gaaaggaacc atgttttgga aggcttactt ggctacatat tatgcactac aatggtatag 240

ctagattcat atgtacttat gatgccctta tattgtttcc tgatgtatta ctcttaaaac 300

cttctttgat caaatttaca ggaggctgca gttgttcctc cctacattgc ctttgccacg 360

agacagaatc ctggattctg ggaatatgtg aaagtgaatg ctaatgatct ttctgttgag 420

ggtattacag ctacagaata cttgaaattc aaggaaatga tagttgatga atgctggtat 480

agtatacgtt gcagcttatc ataccttttg tggttttata acttcaatca gaaaactcat 540

cagagttacc tttgtgtgaa catgaaatgc agggcaaaag atgaatatgc actggaaatt 600

gattttggag cagtagactt ctcaacgcct cgactgaccc tatcctcttc aattggcaat 660

ggtctcagtt atgtttccaa gtttctaact tcaaagctaa atgctacctc cgcgagtgca 720

cagtgtctgg ttgactactt gctcactttg aatcatcaag gagatgtacg tcaacaaaaa 780

tcaaactcca taagtaaact tgtcaactct aagaagaaaa aataggaaaa gaagattcac 840

gtaacaaatt ttctttatgt tcaactgcag aaactgatga tcaatgagac actcagcact 900

gtctcaaagc ttcaggctgc actggttgta gcagaagcat ctatttcctc tttaccaaca 960

gatacaccat atgagagctt tgagctaagg tgatttgttt tttcctctac ttccctccac 1020

ttgtgccatg ctacgtagta ctaagtaact tcaattcttg taaagattca aacagtgggg 1080

ttttgagaaa ggatggggtg atacagctga aagggtcagc gacaccatga gaacactgtc 1140

tgaggtgctt caggcaccag atccattgaa cattcagaag ttctttggaa gggttccaac 1200

tgttttcaat attgtattgt tctctgtcca tggatacttt ggccaagcag atgttcttgg 1260

cttgccagac actggtggtc aggtaagcat ttaatagctt ttacatttaa cttctatgca 1320

ttgacaataa aataattttt aacagtttga ccacttctgc tcttgttcaa caggtagttt 1380

atgttttgga tcaagttgta gcttttgaag aagaaatgct acaaagaatt aaacagcagg 1440

ggctcaatat taagcctcaa attcttgtgg tgagttccta gacaatcgac gtgactatgc 1500

aattatgtag aggctgttta gaaaagttaa tatcatatgt tgattgcaca gttaacccga 1560

ctgattccgg atgcaaaagg aacaaagtgc aaccaggaac tagaaccaat caagaataca 1620

aaacattcac acatcctcag agttccatt aggacagaaa aaggagtgct taatcaatgg 1680

gtttcacgat ttgatatcta tccatatctg gagagatata ctcaggtatg tatttttata 1740

tcaaccttgc tcatcaaaga tgtgttgttt cctcaattcc atttttcccc ttggcaaaag 1800

gatgctgctg acaaaatcgt cgagctaatg gaaggcaaac ctgatctaat cattggtaac 1860

tacactgatg ggaatctagt ggcttcacta atggctagaa aacttgggat aactctggta 1920

acttttctta atcatatttg atgttgcttc ttctccaagt tagttcttaa tctccactga 1980

cctagaccat ctttgcaaca gggaactatt gctcatgctt tggagaagac aaaatatgaa 2040

gactctgaca taaaattgaa ggaactcgat ccgaagtacc acttctcttg ccaattcaca 2100

gctgatttga ttgcaatgaa ttcagcagat ttcattatca ctagcacata ccaagaaata 2160

gctggaaggt aagaattaga gctaataagt aatgcattca tatgtatttc agcatcgctc 2220

tttcaccatc atcgaataca caccactact cagtaaatgt atttgctcaa aagtttgcaa 2280

cttaatggat ctcattcttg aatgcttcaa catatgcagc aaagataaac caggacagta 2340

tgagagccat agtgcattta cccttccagg gctttacaga gttgcttcag gtatcaatgt 2400

ctttgatcca aaatttaata ttgctgcacc tggggcagac cagtcggtgt atttccctta 2460

cacagaaaag cagaagcgtt tgactgcttt ccgccctgcc attgaggaac tgctttttag 2520

taaagtggac aatgacgagc acgtgtaagt ctaagtgtta aacttcagct tagtgcctag 2580

aacatcccac tgctctatgt attgatgttt cacttgtttc aaacagtgga tatttagaag 2640

acagaaagaa acctatcctg tttaccatgg caaggctgga cacagtgaag aacacatctg 2700

gactaacaga atggtatggc aagaacaaga ggctcagaag cttagttaac cttgttgtgg 2760

ttggtggttc ctttgatcct acaaaatcca aggataggga agaagcagct gaaataaaaa 2820

agatgcacat gctgatagag aaataccagc ttaagggtca gattagatgg atagcagctc 2880

agactgacag atacagaaat agtgaactct accgcacaat agcagattcc aaaggagctt 2940

ttgtgcagcc tgcattgtat gaagcatttg gtctaacagt cattgaggca atgaactgtg 3000

gattaccaac ctttgctacc aaccaaggtg gccctgctga gattattgtt gatggggtct 3060

caggctttca tattgatcca aataatgggg atgaatcaag caacaaaatt gccaactttt 3120

tccaaaaatg cagggaggat cctgagtatt ggaacaggat ttcagtccag ggtctaaacc 3180

gtatatatga atggtaactc acagataagc cattcaaatt gcaaagaggc acatatcttg 3240

cagaaaattt cttaatcctt aaatcctaat tttttgcagt tacacatgga agatctatgc 3300

aaacaaggta ttgaatatgg ggtccatcta tactttttgg aggacattgt acagagatca 3360

gaaacaagca aagcaaagat acatcgagac tttctacaat cttgagttta ggaacttggt 3420

atagtgctgc atgacattga cagtatacca caaacatctt tatgagatga attactttta 3480

ataaaattgt ttttaacctt tgcttcctta atggcactta ttgcaggtaa aaaatgtgcc 3540

tatcagaaag gacgaaacac cacaaggacc aaaggagagg gagaaagtta agccacagat 3600

atcacaaagg catgctctaa agcttttgcc tacagttttt caagagaccc tagtatattc 3660

tagtactaaa ttagaattat acagcatgca gcttttgctg ttcacctttc taaatcacca 3720

gttgtgtcaa tcaagttgac aaaatcaata aattgggatt ttccctttcc tatgcttgat 3780

tgttattact cctactttgt ttatggtagt cttccttcat tgttttctcc tgtacttctt 3840

ttactacaac tgtactgaca tactaattat ttctgtgtac caggcgctca caatcaaggt 3900

tgcagaagta agattagata aaattgctac tgcatga 3937

<210> 31

<211> 860

<212> PROTEIN

<213> Nicotiana tabacum

<400> 31

Met Ala Thr Ala Pro Ala Leu Asn Arg Ser Glu Ser Ile Ala Asp Ser

1 5 10 15

Met Pro Glu Ala Leu Arg Gln Ser Arg Tyr His Met Lys Lys Cys Phe

20 25 30

Ala Lys Tyr Ile Glu Gln Gly Lys Arg Met Met Lys Leu His Asn Leu

35 40 45

Met Asp Glu Leu Glu Lys Val Ile Asp Asp Pro Ala Glu Arg Asn His

50 55 60

Val Leu Glu Gly Leu Leu Gly Tyr Ile Leu Cys Thr Thr Met Glu Ala

65 70 75 80

Ala Val Val Pro Pro Tyr Ile Ala Phe Ala Thr Arg Gln Asn Pro Gly

85 90 95

Phe Trp Glu Tyr Val Lys Val Asn Ala Asn Asp Leu Ser Val Glu Gly

100 105 110

Ile Thr Ala Thr Glu Tyr Leu Lys Phe Lys Glu Met Ile Val Asp Glu

115 120 125

Cys Trp Ala Lys Asp Glu Tyr Ala Leu Glu Ile Asp Phe Gly Ala Val

130 135 140

Asp Phe Ser Thr Pro Arg Leu Thr Leu Ser Ser Ser Ile Gly Asn Gly

145 150 155 160

Leu Ser Tyr Val Ser Lys Phe Leu Thr Ser Lys Leu Asn Ala Thr Ser

165 170 175

Ala Ser Ala Gln Cys Leu Val Asp Tyr Leu Leu Thr Leu Asn His Gln

180 185 190

Gly Asp Lys Leu Met Ile Asn Glu Thr Leu Ser Thr Val Ser Lys Leu

195 200 205

Gln Ala Ala Leu Val Val Ala Glu Ala Ser Ile Ser Ser Leu Pro Thr

210 215 220

Asp Thr Pro Tyr Glu Ser Phe Glu Leu Arg Phe Lys Gln Trp Gly Phe

225 230 235 240

Glu Lys Gly Trp Gly Asp Thr Ala Glu Arg Val Ser Asp Thr Met Arg

245 250 255

Thr Leu Ser Glu Val Leu Gln Ala Pro Asp Pro Leu Asn Ile Gln Lys

260 265 270

Phe Phe Gly Arg Val Pro Thr Val Phe Asn Ile Val Leu Phe Ser Val

275 280 285

His Gly Tyr Phe Gly Gln Ala Asp Val Leu Gly Leu Pro Asp Thr Gly

290 295 300

Gly Gln Val Val Tyr Val Leu Asp Gln Val Val Ala Phe Glu Glu Glu

305 310 315 320

Met Leu Gln Arg Ile Lys Gln Gln Gly Leu Asn Ile Lys Pro Gln Ile

325 330 335

Leu Val Leu Thr Arg Leu Ile Pro Asp Ala Lys Gly Thr Lys Cys Asn

340 345 350

Gln Glu Leu Glu Pro Ile Lys Asn Thr Lys His Ser His Ile Leu Arg

355 360 365

Val Pro Phe Arg Thr Glu Lys Gly Val Leu Asn Gln Trp Val Ser Arg

370 375 380

Phe Asp Ile Tyr Pro Tyr Leu Glu Arg Tyr Thr Gln Asp Ala Ala Asp

385 390 395 400

Lys Ile Val Glu Leu Met Glu Gly Lys Pro Asp Leu Ile Ile Gly Asn

405 410 415

Tyr Thr Asp Gly Asn Leu Val Ala Ser Leu Met Ala Arg Lys Leu Gly

420 425 430

Ile Thr Leu Gly Thr Ile Ala His Ala Leu Glu Lys Thr Lys Tyr Glu

435 440 445

Asp Ser Asp Ile Lys Leu Lys Glu Leu Asp Pro Lys Tyr His Phe Ser

450 455 460

Cys Gln Phe Thr Ala Asp Leu Ile Ala Met Asn Ser Ala Asp Phe Ile

465 470 475 480

Ile Thr Ser Thr Tyr Gln Glu Ile Ala Gly Ser Lys Asp Lys Pro Gly

485 490 495

Gln Tyr Glu Ser His Ser Ala Phe Thr Leu Pro Gly Leu Tyr Arg Val

500 505 510

Ala Ser Gly Ile Asn Val Phe Asp Pro Lys Phe Asn Ile Ala Ala Pro

515 520 525

Gly Ala Asp Gln Ser Val Tyr Phe Pro Tyr Thr Glu Lys Gln Lys Arg

530 535 540

Leu Thr Ala Phe Arg Pro Ala Ile Glu Glu Leu Leu Phe Ser Lys Val

545 550 555 560

Asp Asn Asp Glu His Val Gly Tyr Leu Glu Asp Arg Lys Lys Pro Ile

565 570 575

Leu Phe Thr Met Ala Arg Leu Asp Thr Val Lys Asn Thr Ser Gly Leu

580 585 590

Thr Glu Trp Tyr Gly Lys Asn Lys Arg Leu Arg Ser Leu Val Asn Leu

595 600 605

Val Val Val Gly Gly Ser Phe Asp Pro Thr Lys Ser Lys Asp Arg Glu

610 615 620

Glu Ala Ala Glu Ile Lys Lys Met His Met Leu Ile Glu Lys Tyr Gln

625 630 635 640

Leu Lys Gly Gln Ile Arg Trp Ile Ala Ala Gln Thr Asp Arg Tyr Arg

645 650 655

Asn Ser Glu Leu Tyr Arg Thr Ile Ala Asp Ser Lys Gly Ala Phe Val

660 665 670

Gln Pro Ala Leu Tyr Glu Ala Phe Gly Leu Thr Val Ile Glu Ala Met

675 680 685

Asn Cys Gly Leu Pro Thr Phe Ala Thr Asn Gln Gly Gly Pro Ala Glu

690 695 700

Ile Ile Val Asp Gly Val Ser Gly Phe His Ile Asp Pro Asn Asn Gly

705 710 715 720

Asp Glu Ser Ser Asn Lys Ile Ala Asn Phe Phe Gln Lys Cys Arg Glu

725 730 735

Asp Pro Glu Tyr Trp Asn Arg Ile Ser Val Gln Gly Leu Asn Arg Ile

740 745 750

Tyr Glu Cys Tyr Thr Trp Lys Ile Tyr Ala Asn Lys Val Leu Asn Met

755 760 765

Gly Ser Ile Tyr Thr Phe Trp Arg Thr Leu Tyr Arg Asp Gln Lys Gln

770 775 780

Ala Lys Gln Arg Tyr Ile Glu Thr Phe Tyr Asn Leu Glu Phe Arg Asn

785 790 795 800

Leu Val Lys Asn Val Pro Ile Arg Lys Asp Glu Thr Pro Gln Gly Pro

805 810 815

Lys Glu Arg Glu Lys Val Lys Pro Gln Ile Ser Gln Arg His Ala Leu

820 825 830

Lys Leu Leu Pro Thr Val Phe Gln Glu Thr Leu Ala Leu Thr Ile Lys

835 840 845

Val Ala Glu Val Arg Leu Asp Lys Ile Ala Thr Ala

850 855 860

<210> 32

<211> 3905

<212> DNA

<213> Nicotiana tabacum

<400> 32

atggctactg caccagccct gaaaagatca gagtccatag ctgatagcat gccagaggcc 60

ttaaggcaaa gccggtacca catgaagaaa tgttttgcca agtacataga gcaaggcaag 120

aggatgatga aacttcataa cttgatggat gaattggaga aagtaattga tgatcctgct 180

gaaaggaacc atgttttgga aggcttactt ggctacatat tatgtactac aatggtatag 240

ctagattcat atgtacttat gatgtcctta tattgtttcc ggaggcatta ttcttaaatc 300

cttctttgat caaatttgta ggaggctgca gttgttcctc cctatattgc cttcgccacg 360

agacagaatc ctggattctg ggaatatgtg aaagtcaatg ctaatgatct ttctgttgag 420

ggtattacag ctacagatta cttgaaattc aaggaaatga tagttgatga aagctggtat 480

agaatacttt gcagcttatc ataccttttg tggttttata atttcaatca gaaaactcat 540

cagagttacc tttgtgtgaa catgacatgc agggcaaaag atgaatatgc actggaaatt 600

gattttggag cagtagactt ctcaacgcct cgactgaccc tatcctcttc aattggaaat 660

ggtctcagtt atgtttccaa gtttctaact tcaaagctaa atgctacctc agcgagtgca 720

cagtgtctgg ttgactactt gctcactttg aatcaccaag gagatgtacg tcaacaaaaa 780

tcaaactcca taagtaaact tgtcaactct aagaagtaaa aataggaaaa gaagattcat 840

gtaacaaatt ttctttatgt tcaactgtag aaactgatga tcaatgagac actcggcact 900

gtctcaaagc ttcaggctgc actggttgta gcagaagcat ctatttcctc cttaccaaca 960

gatacaccat accagagctt tgagctaagg tgatttgttt tttcctctac ttccttccac 1020

ttttggtgtg ctacatagta ctaagtaact tcaattcttg taaagattca aacagtgggg 1080

ttttgagaaa ggatggggtg atacagctga aagggtccgc gacaccatga gaacactttc 1140

tgaggtactt caggcgccag atccattgaa cattgagaag ttctttggga gggttccaac 1200

tgttttcaat attgtattgt tctctgttca tggatacttt ggccaagcaa atgttcttgg 1260

cttgccagac acaggtggtc aggtaagcat ctaatagctt ttacatttaa cttctatgca 1320

ttgacaataa aataacttct acactaccaa ataatttttg aaagtttgac cacttcggct 1380

cttgttcaac aggtggttta tgttttggat caagttgtag cttttgaaga agaaatgctc 1440

caaagaatta aacagcaggg gctcaatatt aagcctcaaa ttcttgtggt gagctcctag 1500

acaatgacgt gactatgcaa ttaagtagag gctgtttaga aaagttaata tcatatgttg 1560

attgcacagt taacccgact gattccggac gccaaaggaa caaagtgcaa cccaggaacta 1620

gaaccaatca agaatacaaa acattcacac atcctcagag ttccatttag gacagaaaaa 1680

ggagtgctta atcaatgggt ttcacgattt gatatctatc catatctgga gagatatact 1740

caggtgtgta tttttatatc aaccctgctc atcaaagatg tgttgtttcc tcaattccat 1800

ttttcgcctt gacaaaagga cgctgctgac aaaatcatcg agctaatgga aggcaaacct 1860

gatctaatca ttggtaacta cactgatggg aatctagtgg cttctctaat ggctagaaag 1920

cttgggataa ctctggtaac ttttcttatc atatttgatg ttgtttcttc tccaagttgg 1980

ttcttaatgt caactaaccc agaccatctt tgtaacaggg aactattgct catgctctgg 2040

agaagacaaa atatgaagac tctgacatca aattgaagga actcgatccg aagtaccact 2100

tttcttgcca attcacagct gatttgattg caatgaattc agcagatttc attatcacaa 2160

gcacatatca agaaatagcc ggaaggtaag aattggaact acggaagcag agagctaata 2220

agtagtgcac tcatatattt cagcatcgct ctttcgcata atcgaataca caccactact 2280

cagtaaatgt acttgctcaa aagtttacaa gtttatggat cttattcttg aatgcttcaa 2340

catatgcagc aaagataggc caggacagta tgagagccat agtgcattta cccttccagg 2400

gctttacaga gttgcttcag gcatcaatgt ctttgatcct aaatttaata ttgctgcacc 2460

tggggcagac caatcggtgt atttccctta cacagaaaag cagacgcgtt tgactgcttt 2520

ccgccctgcc attgaggaac tgctttttag taaagtggac aatgacgagc acatgtaagt 2580

cttagtgtta aacttcagct ttcagcttag tgcctagaac attccactgg ctctatgtat 2640

taatgtttca cttgtttcaa acacagtgga tatttagaag acagaaagaa acctatcctg 2700

tttaccatgg caaggctgga cacagtgaag aacacatctg gactaacaga atggtatggc 2760

aagaacaaga ggctcagaag cttagttaac cttgttgtgg ttggtggttc ctttgatcct 2820

acaaaatcca aggatagaga agaagcagct gaaataaaaa agatgcacat gctgatagag 2880

aaataccagc ttaagggtca gatcagatgg atagcagctc agactgacag atatagaaac 2940

agtgaactct accgcacaat agcagattcc aaaggagctt ttgtgcagcc tgcattatat 3000

gaagcatttg gtctaacagt cattgaggca atgaactgtg gattaccaac ctttgctacc 3060

aaccaaggtg gccctgctga gattattgtt gatggggtct caggctttca tattgatcca 3120

aataatgggg atgaatcaag caacaaagtt gccaactttt tccaaaaatg cagggaggat 3180

cctgagtatt ggaacaggat ttcagtccag ggtctaaacc gtatatatga atggtaactc 3240

acagataagc cattcaaatt gcaaagaggc acatatcttg ctgaaaattt cttaatcctt 3300

taatcctaaa attttgcagt tacacatgga agatctatgc aaacaaggta ttgaatatgg 3360

ggtccatcta tactttttgg aggacattgt acagagatca gaaacaagca aagcaaagat 3420

acatcgagac tttctacaat cttgagttta ggaacttggt atagtgctgc atgacattga 3480

cagtatacca caaacatctt tatgagatga attactttta ataaaattgt ttttaacctt 3540

tgcctcctta atgacactta ttgcaggtaa aaaatgtgcc tatcagacag gacgaaacac 3600

cacaaggacc aaaggagagg agggagaaag ttaagccaca gatatcacaa aggcatgctc 3660

taaagctttt gcctatagtt tttcaggaga ccctagtata ttctagtact aaattagaat 3720

tatacagcat gcagcttgct tctgctgttc acctttctaa atcaccagtt atgtcaatca 3780

agttgacaaa atcaataaat tcggcttttc cctttcctat gcttgattgt tattactcct 3840

acttcgttta tggtagtctt ccttcattgt tttctcctgt acttctttta ctacaactgt 3900

actga 3905

<210> 33

<211> 913

<212> PROTEIN

<213> Nicotiana tabacum

<400> 33

Met Ala Thr Ala Pro Ala Leu Lys Arg Ser Glu Ser Ile Ala Asp Ser

1 5 10 15

Met Pro Glu Ala Leu Arg Gln Ser Arg Tyr His Met Lys Lys Cys Phe

20 25 30

Ala Lys Tyr Ile Glu Gln Gly Lys Arg Met Met Lys Leu His Asn Leu

35 40 45

Met Asp Glu Leu Glu Lys Val Ile Asp Asp Pro Ala Glu Arg Asn His

50 55 60

Val Leu Glu Gly Leu Leu Gly Tyr Ile Leu Cys Thr Thr Met Glu Ala

65 70 75 80

Ala Val Val Pro Pro Tyr Ile Ala Phe Ala Thr Arg Gln Asn Pro Gly

85 90 95

Phe Trp Glu Tyr Val Lys Val Asn Ala Asn Asp Leu Ser Val Glu Gly

100 105 110

Ile Thr Ala Thr Asp Tyr Leu Lys Phe Lys Glu Met Ile Val Asp Glu

115 120 125

Ser Trp Ala Lys Asp Glu Tyr Ala Leu Glu Ile Asp Phe Gly Ala Val

130 135 140

Asp Phe Ser Thr Pro Arg Leu Thr Leu Ser Ser Ser Ile Gly Asn Gly

145 150 155 160

Leu Ser Tyr Val Ser Lys Phe Leu Thr Ser Lys Leu Asn Ala Thr Ser

165 170 175

Ala Ser Ala Gln Cys Leu Val Asp Tyr Leu Leu Thr Leu Asn His Gln

180 185 190

Gly Asp Lys Leu Met Ile Asn Glu Thr Leu Gly Thr Val Ser Lys Leu

195 200 205

Gln Ala Ala Leu Val Val Ala Glu Ala Ser Ile Ser Ser Leu Pro Thr

210 215 220

Asp Thr Pro Tyr Gln Ser Phe Glu Leu Arg Phe Lys Gln Trp Gly Phe

225 230 235 240

Glu Lys Gly Trp Gly Asp Thr Ala Glu Arg Val Arg Asp Thr Met Arg

245 250 255

Thr Leu Ser Glu Val Leu Gln Ala Pro Asp Pro Leu Asn Ile Glu Lys

260 265 270

Phe Phe Gly Arg Val Pro Thr Val Phe Asn Ile Val Leu Phe Ser Val

275 280 285

His Gly Tyr Phe Gly Gln Ala Asn Val Leu Gly Leu Pro Asp Thr Gly

290 295 300

Gly Gln Val Val Tyr Val Leu Asp Gln Val Val Ala Phe Glu Glu Glu

305 310 315 320

Met Leu Gln Arg Ile Lys Gln Gln Gly Leu Asn Ile Lys Pro Gln Ile

325 330 335

Leu Val Leu Thr Arg Leu Ile Pro Asp Ala Lys Gly Thr Lys Cys Asn

340 345 350

Gln Glu Leu Glu Pro Ile Lys Asn Thr Lys His Ser His Ile Leu Arg

355 360 365

Val Pro Phe Arg Thr Glu Lys Gly Val Leu Asn Gln Trp Val Ser Arg

370 375 380

Phe Asp Ile Tyr Pro Tyr Leu Glu Arg Tyr Thr Gln Asp Ala Ala Asp

385 390 395 400

Lys Ile Ile Glu Leu Met Glu Gly Lys Pro Asp Leu Ile Ile Gly Asn

405 410 415

Tyr Thr Asp Gly Asn Leu Val Ala Ser Leu Met Ala Arg Lys Leu Gly

420 425 430

Ile Thr Leu Gly Thr Ile Ala His Ala Leu Glu Lys Thr Lys Tyr Glu

435 440 445

Asp Ser Asp Ile Lys Leu Lys Glu Leu Asp Pro Lys Tyr His Phe Ser

450 455 460

Cys Gln Phe Thr Ala Asp Leu Ile Ala Met Asn Ser Ala Asp Phe Ile

465 470 475 480

Ile Thr Ser Thr Tyr Gln Glu Ile Ala Gly Ser Lys Asp Arg Pro Gly

485 490 495

Gln Tyr Glu Ser His Ser Ala Phe Thr Leu Pro Gly Leu Tyr Arg Val

500 505 510

Ala Ser Gly Ile Asn Val Phe Asp Pro Lys Phe Asn Ile Ala Ala Pro

515 520 525

Gly Ala Asp Gln Ser Val Tyr Phe Pro Tyr Thr Glu Lys Gln Thr Arg

530 535 540

Leu Thr Ala Phe Arg Pro Ala Ile Glu Glu Leu Leu Phe Ser Lys Val

545 550 555 560

Asp Asn Asp Glu His Ile Gly Tyr Leu Glu Asp Arg Lys Lys Pro Ile

565 570 575

Leu Phe Thr Met Ala Arg Leu Asp Thr Val Lys Asn Thr Ser Gly Leu

580 585 590

Thr Glu Trp Tyr Gly Lys Asn Lys Arg Leu Arg Ser Leu Val Asn Leu

595 600 605

Val Val Val Gly Gly Ser Phe Asp Pro Thr Lys Ser Lys Asp Arg Glu

610 615 620

Glu Ala Ala Glu Ile Lys Lys Met His Met Leu Ile Glu Lys Tyr Gln

625 630 635 640

Leu Lys Gly Gln Ile Arg Trp Ile Ala Ala Gln Thr Asp Arg Tyr Arg

645 650 655

Asn Ser Glu Leu Tyr Arg Thr Ile Ala Asp Ser Lys Gly Ala Phe Val

660 665 670

Gln Pro Ala Leu Tyr Glu Ala Phe Gly Leu Thr Val Ile Glu Ala Met

675 680 685

Asn Cys Gly Leu Pro Thr Phe Ala Thr Asn Gln Gly Gly Pro Ala Glu

690 695 700

Ile Ile Val Asp Gly Val Ser Gly Phe His Ile Asp Pro Asn Asn Gly

705 710 715 720

Asp Glu Ser Ser Asn Lys Val Ala Asn Phe Phe Gln Lys Cys Arg Glu

725 730 735

Asp Pro Glu Tyr Trp Asn Arg Ile Ser Val Gln Gly Leu Asn Arg Ile

740 745 750

Tyr Glu Cys Tyr Thr Trp Lys Ile Tyr Ala Asn Lys Val Leu Asn Met

755 760 765

Gly Ser Ile Tyr Thr Phe Trp Arg Thr Leu Tyr Arg Asp Gln Lys Gln

770 775 780

Ala Lys Gln Arg Tyr Ile Glu Thr Phe Tyr Asn Leu Glu Phe Arg Asn

785 790 795 800

Leu Val Lys Asn Val Pro Ile Arg Gln Asp Glu Thr Pro Gln Gly Pro

805 810 815

Lys Glu Arg Arg Glu Lys Val Lys Pro Gln Ile Ser Gln Arg His Ala

820 825 830

Leu Lys Leu Leu Pro Ile Val Phe Gln Glu Thr Leu Val Tyr Ser Ser

835 840 845

Thr Lys Leu Glu Leu Tyr Ser Met Gln Leu Ala Ser Ala Val His Leu

850 855 860

Ser Lys Ser Pro Val Met Ser Ile Lys Leu Thr Lys Ser Ile Asn Ser

865 870 875 880

Ala Phe Pro Phe Pro Met Leu Asp Cys Tyr Tyr Ser Tyr Phe Val Tyr

885 890 895

Gly Ser Leu Pro Ser Leu Phe Ser Pro Val Leu Leu Leu Leu Gln Leu

900 905 910

Tyr

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Claims (40)

1. A tobacco plant cell capable of expressing the INV gene, containing: (i) a polynucleotide comprising, consisting or consisting essentially of a sequence characterized by at least 81% sequence identity to SEQ ID NO: 5 ( NtINV4-S ) or at least 85% sequence identity to SEQ ID NO: 7 ( NtINV4-T ); (ii) a polypeptide encoded by the polynucleotide specified in (i); (iii) a polypeptide comprising, consisting or consisting essentially of a sequence having at least 85% sequence identity to SEQ ID NO: 6 (NtINV4-S) or at least 85% sequence identity to SEQ ID NO: 8 (NtINV4-T); or (iv) a construct, vector or expression vector comprising the isolated polynucleotide referred to in (i), wherein the tobacco plant cell comprises at least one modification that provides for a decrease in (a) the expression or activity of the polynucleotide or (b) the expression or activity of the polypeptide encoding the polypeptide, compared to a control tobacco plant cell in which the expression or activity of the polynucleotide or polypeptide has not been modified, wherein the reduced expression or reduced activity provides for a reduced level of one or more reducing sugars in a dried leaf of a plant containing a cell of a tobacco plant, compared to a level of one or more reducing sugars in a dried leaf of a control plant containing a cell of a control tobacco plant, wherein preferably the reducing sugar is glucose or fructose, or a combination thereof. 2. The tobacco plant cell of claim 1, wherein the dried leaf obtained from the midsection of the plant has reduced glucose levels of at least about 63% compared to a control dried leaf obtained from the midsection of the plant; or the dried leaf obtained from the midsection of the plant is characterized by reduced fructose levels of at least about 43%, respectively, compared to a control dried leaf obtained from the midsection of the plant; or the dried leaf obtained from the midsection of the plant is characterized by reduced glucose levels of at least about 63% and reduced fructose levels of at least about 43%, respectively, compared to a control dried leaf obtained from the midsection of the plant. 3. A tobacco plant cell according to claim 1 or 2, wherein at least one modification is a genetic mutation in a polynucleotide and the plant is Nicotiana tabacum . 4. A tobacco plant cell according to any of the preceding claims, further comprising at least one modification in the NtSUS polynucleotide or the polypeptide encoded by it, wherein preferably the NtSUS polynucleotide or the polypeptide encoded by it is selected from the group consisting of NtSUS2-T , NtSUS3-S , NtSUS3-T , NtSUS4-S , NtSUS4-T or a combination of two or more of them, wherein more preferably the NtSUS polynucleotide or the polypeptide encoded therefrom is selected from the group consisting of NtSUS2-S , NtSUS3-S , NtSUS3-T and NtSUS4-S or a combination of two or more of them. 5. A tobacco plant cell according to claim 4, comprising at least one mutation in the NtINV4 polynucleotide(s) specified in (i) of claim 1, or the NtINV4 polypeptide(s) specified in (ii) of claim 1 or in (iii) of claim 1, and at least one mutation in the NtSUS polynucleotide(s) or the polypeptide(s) encoded by it. 6. A tobacco plant having a reduced content of reducing sugars formed during drying, wherein the plant comprises a tobacco plant cell according to any one of paragraphs 1-5. 7. A part of a tobacco plant having a reduced content of reducing sugars formed during drying, wherein the part of the plant comprises a cell of a tobacco plant according to any one of paragraphs 1-5. 8. Tobacco plant material, dried tobacco plant material or homogenized tobacco plant material having a reduced content of reducing sugars formed during drying and containing a tobacco plant cell according to any one of claims 1-5. 9. The tobacco plant material, dried tobacco plant material or homogenized tobacco plant material of claim 8, wherein the tobacco plant material is selected from the group consisting of biomass, seed, stem, flowers or leaves or a combination of two or more thereof; or the dried plant material is selected from the group consisting of flue-dried plant material, sun-dried plant material or air-dried plant material, or a combination of two or more thereof. 10. A tobacco product having a reduced content of reducing sugars formed during drying and containing a tobacco plant cell according to any one of claims 1-5, a part of a plant according to claim 7 containing a tobacco plant cell according to any one of claims 1-5, or a tobacco plant material according to claim 8 containing a tobacco plant cell according to any one of claims 1-5. 11. A method for obtaining a tobacco plant according to claim 6, comprising the following stages: (a) obtaining a tobacco plant cell comprising at least one modification according to any one of paragraphs 1 to 5; and (b) reproducing a tobacco plant cell to produce a tobacco plant. 12. The method according to claim 11, wherein in step (a) at least one modification is introduced by genome editing. 13. The method of claim 11, wherein the genome editing methods are selected from CRISPR-mediated genome editing, zinc finger nuclease-mediated mutagenesis, chemical or radiation mutagenesis, homologous recombination, oligonucleotide-directed mutagenesis, and meganuclease-mediated mutagenesis. 14. The method according to claim 11 or 12, wherein in step (a) at least one modification is introduced using a polynucleotide to provide interference or by introducing at least one mutation or a combination thereof. 15. A method for obtaining dried plant material with a reduced amount of reducing sugars compared to a control plant material, comprising the steps of: (a) obtaining a plant or part thereof according to paragraph 6 or 7 or plant material according to paragraph 8 or 9; (b) collecting plant material from them and (c) drying of plant material. 16. A method for producing liquid tobacco extract, comprising the following stages: (a) obtaining a first tobacco starting material from a plant or part thereof comprising a tobacco plant cell according to any one of paragraphs 1 to 3; (b) obtaining a second tobacco starting material from a plant or part thereof comprising a tobacco plant cell comprising at least one modification in the NtSUS polynucleotide or the polypeptide encoded by it, (c) heating the first tobacco starting material at a first extraction temperature; (d) heating the second tobacco feedstock material at a second extraction temperature; (e) collecting volatile compounds released from the first tobacco starting materials and the second tobacco starting materials during heating; and (f) combining the collected volatile compounds released from the first and second tobacco starting materials and forming a liquid tobacco extract from the combined volatile compounds. 17. The method of claim 16, wherein the NtSUS polynucleotide or the polypeptide encoded by it in step (b) is selected from the group consisting of NtSUS2-T , NtSUS3-S , NtSUS3-T , NtSUS4-S , NtSUS4-T or a combination of two or more of them. 18. The method of claim 16, wherein the NtSUS polynucleotide or the polypeptide encoded by it in step (b) is selected from the group consisting of NtSUS2-S , NtSUS3-S , NtSUS3-T and NtSUS4-S or a combination of two or more of them.

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