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EP1326999A2 - Procede permettant la prediction des phenotypes d'arbres a l'aide de l'acide nucleique - Google Patents

Procede permettant la prediction des phenotypes d'arbres a l'aide de l'acide nucleique

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Publication number
EP1326999A2
EP1326999A2 EP01949143A EP01949143A EP1326999A2 EP 1326999 A2 EP1326999 A2 EP 1326999A2 EP 01949143 A EP01949143 A EP 01949143A EP 01949143 A EP01949143 A EP 01949143A EP 1326999 A2 EP1326999 A2 EP 1326999A2
Authority
EP
European Patent Office
Prior art keywords
tree
trees
restriction
hybrid
nucleic acid
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP01949143A
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German (de)
English (en)
Inventor
Simon Potter
Paul A. Watson
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Pulp and Paper Research Institute of Canada
Original Assignee
Pulp and Paper Research Institute of Canada
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Filing date
Publication date
Application filed by Pulp and Paper Research Institute of Canada filed Critical Pulp and Paper Research Institute of Canada
Publication of EP1326999A2 publication Critical patent/EP1326999A2/fr
Withdrawn legal-status Critical Current

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Classifications

    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
    • A01H1/04—Processes of selection involving genotypic or phenotypic markers; Methods of using phenotypic markers for selection
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H7/00—Gymnosperms, e.g. conifers
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6813—Hybridisation assays
    • C12Q1/6827—Hybridisation assays for detection of mutation or polymorphism
    • C12Q1/683—Hybridisation assays for detection of mutation or polymorphism involving restriction enzymes, e.g. restriction fragment length polymorphism [RFLP]
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
    • C12Q1/6895—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms for plants, fungi or algae
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/13—Plant traits
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/156—Polymorphic or mutational markers

Definitions

  • This invention is in the fields of molecular biology and tree improvement, and pulp and paper properties evaluations. This invention allows for an enhanced efficiency of selection for trees with given fibre properties from both natural and plantation populations for specific pulp and paper product lines.
  • Probes based on nuclear sequences have also been used with varying success - for example, riboso al DNA (rDNA) regions of red and black spruce are known to exhibit intraspecies variation of some polymorphic regions which have been exploited to produce species- specific RAPD markers. By their nature, these markers are based on arbitrary sequences whose general applicability (i.e. across species) is difficult to determine (Bobola, M.S. et al.,(1992j Mol . Biol . Evol . 9:125-137), Perron, M. et al . , (1997) Molecular ecology 6:725-734) .
  • the present invention describes a nucleic acid probe developed for interior / Sitka hybrids (in the Nass-Skeena introgression zone) which is capable of quantitatively estimating the degree of genetic hybridization between the two.
  • This probe Eco2.0, is based on a stretch of yeast (Saccharo yces cerevisiae) 18S rDNA and detects species-specific 18S external transcribed spacer (ETS) polymorphisms in a quantitative fashion as determined by densitometric image analysis.
  • One aim of the present invention is to provide a method for identifying individual trees having superior phenotype .
  • a Eco2.0 probe a Eco2.0 probe, its purification and sequence characterization, and and its use as a marker for the degree of introgression in individual hybrids within the Nass-Skeena zone.
  • the Eco2.0 probe has been isolated and cloned into E. coli
  • the probe has been purified and partially sequenced to determine it's identity.
  • the utility of the probe as a marker for the fibre quality of the hybrids has been assessed by a direct comparison of Southern blot band intensity patterns to fibre property parameters for individual tree samples selected' from geographical areas spanning the Nass-Skeena introgression zone.
  • a method for identifying tree lineage capable of expressing desired biological and/or biochemical phenotypes comprising the steps of: a) obtaining a nucleic acid sample, such as RNA or DNA, from trees of pure species and/or hybrid thereof ; b) obtaining a restriction pattern of restriction fragments by subjecting said nucleic acid sample of step a) to at least one restriction enzyme, wherein said restriction enzyme maximizes differences between said restriction pattern of pure species and/or hybrid thereof; c) visualizing said restriction pattern of step b) by submitting the treated nucleic acid sample of step b) to at least one labeled probe, such as without limitation Eco2.0 , for complementary hybridization between said probe and said nucleic acid sample, wherein said probe allows for detection of the degree of hybridization and/or different intensity of said restriction fragments between trees of pure species and/or hybrid thereof; and d) correlating said restriction pattern and/or intensity of restriction fragments of step c) to at least one selected biological and biochemical
  • Step d) may further comprise a standard curve for predictive relationship between ' said restriction pattern and/or intensity of restriction fragments, and said phenotype .
  • the tree of pure species and/or hybrid thereof may be naturally or artificially produced.
  • the nucleic acid sample of step a) may be obtained from a leaf, cambium, root, bud, stem, cork, phloem or xylem.
  • the tree may be of the genus Picea, or may be of a genus selected from the group consisting of Populus, Be tula, Abies, Larix, Taxus, Ulmus, Prunus, Quercus, Malus, Arbutus, Salix, Platanus, Acer, Tsuga, Pseudotsuga, Pinus, Fraxinus, Eucalyptus, Acacia, Abrus, Cupressus, Fagus, Juniperus, Thuja, and Canya .
  • Step c) may further comprise measurement of intensity of said restriction fragments.
  • the biological or biochemical phenotype searched for may be for example selected from the group consisting of fiber length, wood density, fiber collapsibility, fiber coarseness, cell wall thickness, growth rate, lignin content, guaiacyl lignin content, syringyl lignin content, carbohydrate ' content, kraft pulp yield, mechanical pulp energy demand, chemical uptake for chemical pulping, extractive content, and extractive compounds.
  • a pattern of restriction fragments obtained by subjecting a nucleic acid sample from trees of pure species and/or hybrid thereof to at least one restriction enzyme for identifying tree lineage capable of expressing desired biological and/or biochemical phenotypes, wherein said restriction enzyme maximizes differences between said pattern of restriction fragments from pure species and/or hybrid thereof.
  • a pattern of restriction fragments obtained by subjecting a nucleic acid sample from trees of pure species and/or hybrid thereof to at least one restriction enzyme for use in identifying tree lineage capable of expressing desired biological and/or biochemical phenotypes, wherein said restriction enzyme maximizes differences between said pattern of restriction fragments from pure species and/or hybrid thereof.
  • a method of screening a plurality of trees of diverse phenotypes which comprises the steps of : a) Characterizing wood quality of at least two trees with different degree of hybridization; b) Developing a standard curve from said trees for a predictive relationship between restriction patterns and phenotypes; c) assessing a plurality of natural species hybrids for restriction patterns of a plurality of hybridization markers; d) comparing said restriction patterns of said hybrids with said standard curve to deduce a phenotype of said hybrids; and e) harvesting said hybrids based on predicted phenotypes .
  • a method of producing a plurality of clonal trees having predictable, consistent and/or enhanced wood or fibre quality properties comprises the steps of : a) characterizing wood quality of at least two trees with different degree of hybridization; b) developing a standard curve from said trees for a predictive relationship between restriction patterns and phenotypes; c) assessing a plurality of natural species hybrids for restriction patterns of a plurality of hybridization markers; d) comparing said restriction patterns of said hybrids with said standard curve to deduce a phenotype of said hybrids; e) obtaining a plurality of progeny trees from said parental trees by performing cross-pollination or somatic embryogenesis; and propagating somatic embryos of said progeny trees obtained in step e) to produce a plurality of clonal trees, essentially all of said clonal trees having predictable, consistent and/or enhanced wood or fibre quality properties.
  • the present invention further provides a stand of clonal trees with enhanced wood or fibre properties produced by the method of the present invention, the genome of said trees containing a restriction pattern, said restriction pattern being the same restriction pattern associated with said enhanced wood or fibre properties.
  • Eco2.0 probe may also be used as a marker for predicting fiber quality of tree samples of pure species and/or hybrid thereof.
  • locus is intended to mean the position occupied on the chromosome by the gene representing a particular trait.
  • restriction fragment length polymorphism means a restriction map that identifies a linear series of sites in DNA, separated from one another by actual distance along the nucleic acid sequence.
  • a restriction map can be obtained for any sequence of DNA, irrespective of whether mutations have been identified in it, or, indeed, whether we have any knowledge of its function.
  • a difference in restriction maps between two individuals can be used as a genetic marker in exactly the same way as any other marker. To relate the restriction map to the genetic map, we must compare the restriction maps of wild-type and corresponding variant, or phenotypes.
  • restriction fragments means fragments of nucleic acid generated following digestion of a purified nucleic acid with restriction enzymes. Restriction fragments may be considered as restriction markers, that are not restricted to those changes that affect a phenotype, they provide the basis for an extremely powerful technique for identifying genetic loci at the molecular level.
  • hybridization means the ability of two separated complementary strands to reform into a double helix. Renaturation depends on specific base pairing between the complementary strands. The reaction takes place in two stages. First, DNA single strands in the solution encounter one another by chance; if their sequences are complementary, the two strands base pair to generate a short double-helical region. Then- the region of base pairing extends along the molecule be a zipper-like effect to form a lengthy duplex molecule. Renaturation describes the reaction between two complementary sequences that were separated by denaturation. However, the technique can be extended to allow any two complementary nucleic acid sequences to anneal with each other to form a duplex structure.
  • the reaction is generally described as hybridization when nucleic acids from different sources are involved, as in the case when one preparation consists of DNA and one of RNA.
  • the ability of two nucleic acids preparations to hybridize constitutes a precise test -for their sequence complementarity.
  • predictive relationship means a correlating factor based on a standard curve established between i) a restriction pattern and/or intensity of restriction fragments, and a particular phenotype; or ii) a "random amplified polymorphic DNA” (RAPD) pattern and a particular phenotype .
  • RAPD random amplified polymorphic DNA
  • hybrid thereof means a progeny issued from the interbreeding of trees of different breeds, varieties, species, especially as produced through tree crossbreeding for specific genetic and phenotypic characteristics. A hybrid thereof is derived by crossbreeding two different tree species .
  • Fig. 1 illustrates a map of British Columbia spruce species distribution showing the Nass Skeena Transition introgression zone
  • Fig. 2 illustrates a map of sampling sites across the Introgression zone. Black and grey circles show sampling sites taken for previous study. Red circles indicate sites sampled for the present study;
  • Fig. 3A illustrates the pEco2.0 plasmid map. Multiple cloning site region is exploded to show the precise ordering of restriction sites. Insertion point for the 2kb yeast rDNA probe is indicated;
  • Fig. 3B illustrates an agarose gel analysis of the plasmid and probe insert . Lanes are described in the text. C: Agarose gel analysis of PCR products using M13 F/R primers. Lanes L to R: markers, PCR products using different proprietary buffers;
  • Fig. 4 illustrates the sequence of the 2kb yeast rDNA probe.
  • Bold sequence represents the probe, ends of which were determined and found to be identical to Genbank Z73326;
  • Fig. 5 illustrates the sequence analysis of the Eco2.0 plasmid using M13 r and f primers
  • Fig. 6 illustrates: Panel B - Southern banding pattern and densitometric analysis of the hybridization of. the probe to total interior and Sitka spruce hybrid DNA digests. Lanes are described in the text. * Sample from site 2 underloaded. In the text, the bands seen are referred to as Bands 1 - 5 from the top down. Panels A and- C show typical banding patterns for the pure species and densitometry of those patterns;
  • Fig. 7 illustrates fibre properties as a function of tree age.
  • the 60-80 age class was chosen for comparative purposes as this is the region free from juvenile wood for all samples;
  • Fig. 8 illustrates fibre coarseness versus length-weighted fibre length showing a strong positive correlation
  • Fig. 9 illustrates a scatterplot matrix for fibre properties, site index and band intensities data.
  • Fig. 10 illustrates the relationship between Band intensity and length-weighted fibre length (LWFL) in the spruce hybrids and indicates an excellent linear relationship between Band 2 and LWFL;
  • Fig. 11 illustrates a standard curve for relationship between relative Band 2 intensity and length-weighted fibre length obtained from Blot 1 (Fig. 5) . Dotted lines show the Band 2 intensity values for spruce samples probed on Blot 2 used in the predictive analysis for fibre length;
  • Fig. 12 illustrates the second hybridization analysis (Blot 2) used for fibre length prediction experiment. Band 2 is indicated. Lane 1, pEco2.0 plasmid insert positive control; lane 2, Nass-Skeena "Black spruce” sample; lanes 3-8, trees 7-9, 6-4, 5-8, 4-5, 2-4; and
  • Fig. 13 illustrates that fibre length as a function of site productivity shows no global correlation. Specific biogeoclimatic zones are delineated to emphasize possible intra-zonal correlations.
  • a novel method for the prediction of selected phenotypes and the rapid selection of superior trees for given pulp and paper product lines using a DNA probe comprises the isolation of tree genomic DNA from a spruce live tissue source, hybridization of the DNA probe to that genomic DNA and the densitometric assessment of the intensity of the hybridization pattern obtained.
  • a particular embodiment of the invention is the determination of the precise degree of genetic admixing (or introgression) of the two parent species within the hybrid population. Due to the linear relationship - in the hybrid spruce population for example - between degree of genetic introgression and fibre length (with regard to the present invention) , the intensity of the DNA probe hybridization pattern is used to directly, accurately and reproducibly predict the fibre length found (for a given tree age) within an individual hybrid Sitka/Interior spruce within the population.
  • a preferred embodiment of the invention is the use of DNA probes as predictive tools in forestry tree improvement programs as it represents the first successful practical demonstration of such an application.
  • One embodiment of the present invention is to provide a novel technique for the rapid assessment of fibre quality in all forest species.
  • Another particular embodiment of the invention is the use of the invention on hybrid spruce specifically, although the invention is likely usable for all forest species.
  • One embodiment of the method invention described herein is related to DNA-based tests - for all wood quality parameters important for the desirability of wood for various solid wood and pulp and paper product applications - which enables acceleration of the process of assessment of natural populations and enables the early selection of elite lines for plantation establishment.
  • the nucleic acid-based tests for difficult and expensive to measure traits provides a highly increased capability for cost reduction and product enhancement in selecting superior tree families.
  • the cones of the hybrid species display characteristics of their parent species, but true species identification and degree of hybridization is difficult to determine by such gross methods .
  • site quality average or typical sites for the species were
  • biogeoclimatic ecosytem classification (BEC) method for site identification and analysis utilizes vegetation, soils, climate, and topography in classifying ecosystems and estimating site quality [Banner, A. et al . , (1993), A field guide to site identification and interpretation for the Prince Rupert forest region. B.C. Ministry of Forests, Victoria, B.C. Land management handbook No. 26) .
  • Site series was determined from estimated soil moisture and soil nutrient regimes. This was then related to site productivity (SIBEC) by estimating site index.
  • Site index - a measure of the productivity of the stand - provides a standardized comparison of the productive potential between sites and is also determined through the site index curve method, which is a measure of tree height at fifty years breast height age (How to determine site index in silviculture. Participants work book, B.C. Ministry of forests, Forest renewal B.C. (1998) ) .
  • Spruce fibre properties were determined from the 10 mm increment core samples. At least one sample from each site was sectioned into age classes to monitor changes in fibre properties over time and to determine the juvenile/mature wood transition point for each of the species / hybrids studied. For all other samples only the 60-80 yr. age class was examined, as this was determined to be in the mature wood zone for each site.
  • Fibres were released from the samples using a hydrogen peroxide / acetic acid maceration technique (Burkart, L.F. (1966) For. Prod. J. 16:52) and the resultant pulps were analyzed using an automated Fibre Quality Analyzer instrument to determine length weighted fibre lengths (LWFL) and fibre coarsenesses according to previously described protocols [Morrison, D. et al . , (1998)Paprican PPR 1403).
  • Genomic DNA from hybrids and pure Sitka and interior spruce was obtained using a FastPrep instrument (Bio 101 Inc.) according to the manufacturers standard protocols.
  • the Eco2.0 rDNA probe-containing plasmid (pEco2.0 - obtained courtesy of CH. Newton, B.C. Research Inc., Vancouver) was isolated, purified and cloned into E. coli according to methods described in Sambrook et al . (Sambrook, J. et al., (1990) Molecular Cloning. A laboratory manual, 2 nd edition, Cold Spring Harbor Laboratory Press) . Restriction Digests
  • Both spruce genomic DNA samples and pEco2.0 were each digested with the appropriate restriction enzymes (HinD III and Eco R I respectively) according to the method of Hanish and McClelland (Hanish, J. et al . , (1988) Gene Anal . Tech . 5:105).
  • Approximately 25 ng of DNA was incubated at 37°C in KGB buffer (1 M potassium glutamate, 250 mM Tris acetate pH 7.5, 100 mM magnesium acetate, 0.5 mg/ml BSA fraction 5, 5 mM ⁇ - mercaptoethanol) with 1 U of restriction enzyme for 2 hr and the reaction stopped with 0.5 M EDTA.
  • Digests were analyzed on 1% agarose gels according to the method described in Sambrook et al . , (1990) . • Polymerase Chain Reaction
  • Terminal sequencing of the pEco2.0 plasmid was performed according to standard dideoxy-nucleotide termination protocols by the Nucleic Acid and Protein Services of the Biotechnology Laboratory at University of British Columbia. Since pEco2.0 is based on the pUC 18 vector, standard M13 forward and reverse primers were used for sequence priming.
  • Fig. 3A shows a schematic of the pUC 18 - based plasmid pEco2.0 and the insertion point of the probe into the multiple cloning site (MCS) .
  • MCS multiple cloning site
  • FIG. 3C shows a PCR amplification experiment using the pEco2.0 plasmid as template and M13 forward and reverse primers.
  • Fig. 4 presents a fragment of Saccharomyces cerevisiae chromosome XII cosmid reading frame ORF YLR154C taken from genbank acc# Z73326. The complete sequence of the Eco2.0 probe is highlighted in bold face.
  • Fig. 5 shows sequence analysis results for the pEco2.0 plasmid obtained using M13 forward and reverse primer . Characterization of Sitka / Interior Spruce rDNA Polymorphisms
  • the pEco2.0 rDNA probe detects a polymorphic ' region of spruce DNA which . is quite distinct in interior and Sitka species.
  • the interior sample gives five bands of varying intensity after hybridization with the probe, whereas the Sitka sample gives only three.
  • Bands 4 and 5 which are diagnostic of Sitka spruce, can be used in conjunction with chloroplast and mitochondrion - specific DNA markers to unambiguously differentiate between the species.
  • Bands 1, 2 and 3 however are common to both species but are present in different relative intensities (Fig. 6) . Densitometry of Band 2 was then used to estimate degrees of introgression exhibited by the hybrid trees sampled across the zone.
  • Lane 2 contains genomic DNA isolated from a tree identified morphologically as a Black spruce sample obtained from a site at Nass-Skeena. This DNA, when tested with the pEco2.0 probe, gives a pattern of bands identical to that seen for interior spruce samples from B.C., suggesting that the probe may potentially also be able to distinguish hybrids of other spruce species.
  • Fibre length 60 - 80 yr is plotted as a function of site index in Fig. 13. It is evident that there is no correlation between site productivity and fibre length across biogeoclimatic zones, indicating that the genetic differences between these hybrids mask any significant macroenvironmental effect.

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Abstract

La présente invention concerne un nouveau procédé permettant la prédiction de la longueur de fibre et la sélection rapide d'arbres supérieurs pour des lignes données de produits de pulpe et de papier grâce à une sonde ADN. Le procédé comprend l'isolation d'ADN génomique d'arbres à partir d'une source de tissu vivant d'une épinette hybride, l'hybridation de la sonde ADN de l'épinette avec cet ADN génomique et l'évaluation densitométrique de l'intensité du modèle d'hybridation obtenu. On détermine ainsi le degré précis de mélange génétique (ou d'introgression) des deux espèces parentes au sein de la population hybride. Grâce à la relation linéaire (découverte dans ce procédé) - dans la population d'épinettes hybrides examinées - entre le degré d'introgression génétique et la longueur de fibre, l'intensité du modèle d'hybridation de la sonde ADN peut être utilisée afin de prédire de manière directe, précise et reproductible la longueur de fibre trouvée (pour un âge donné de l'arbre) pour une épinette hybride individuelle au sein de la population.
EP01949143A 2000-06-23 2001-06-21 Procede permettant la prediction des phenotypes d'arbres a l'aide de l'acide nucleique Withdrawn EP1326999A2 (fr)

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US213585P 2000-06-23
PCT/CA2001/000927 WO2002004663A2 (fr) 2000-06-23 2001-06-21 Procede permettant la prediction des phenotypes d'arbres a l'aide de l'acide nucleique

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US20110296753A1 (en) * 2010-06-03 2011-12-08 Syngenta Participations Ag Methods and compositions for predicting unobserved phenotypes (pup)
CN102676649B (zh) * 2012-02-29 2014-06-04 西北大学 白皮松父系鉴定方法
CN103279636B (zh) * 2013-04-10 2016-03-02 北京林业大学 一种确定树木年龄的方法
CN105274103B (zh) * 2015-11-22 2018-05-25 保琦蓓 一种鉴别汉麻、苎麻、亚麻原麻纤维种类的pcr引物组以及pcr鉴别方法
CN106811513B (zh) * 2015-12-01 2020-12-01 中华人民共和国上海出入境检验检疫局 桉树成分实时荧光pcr检测方法及其试剂盒
CN108467900B (zh) * 2017-02-23 2020-03-10 北京林业大学 一种联合利用lncRNA及其靶基因筛选杨树生长性状的方法、试剂盒及应用
CN109727640B (zh) * 2019-01-22 2021-03-02 隆平农业发展股份有限公司 基于自动机器学习技术的全基因组预测方法及装置
CN114486416A (zh) * 2021-12-24 2022-05-13 华东师范大学 一种基于植物经济型谱的腐木分解速率的检测方法

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AU1682595A (en) * 1994-01-21 1995-08-08 North Carolina State University Methods for within family selection in woody perennials using genetic markers

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WO2002004663A2 (fr) 2002-01-17

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