[go: up one dir, main page]

WO2025140990A1 - Gène conférant une résistance contre peronospora farinosa dans l'épinard - Google Patents

Gène conférant une résistance contre peronospora farinosa dans l'épinard Download PDF

Info

Publication number
WO2025140990A1
WO2025140990A1 PCT/EP2024/087997 EP2024087997W WO2025140990A1 WO 2025140990 A1 WO2025140990 A1 WO 2025140990A1 EP 2024087997 W EP2024087997 W EP 2024087997W WO 2025140990 A1 WO2025140990 A1 WO 2025140990A1
Authority
WO
WIPO (PCT)
Prior art keywords
spinach
spinach plant
cdna
resistance
seq
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.)
Pending
Application number
PCT/EP2024/087997
Other languages
English (en)
Inventor
Raimon Jozef LAAN
Klaas SCHOTANUS
Iris Lisa HAVERKORN
Joëlle SCHACHTSCHABEL
Roelof Marinus Veenstra
Albertus Johannes Maria Schrijver
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.)
Bejo Zaden BV
Original Assignee
Bejo Zaden BV
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Bejo Zaden BV filed Critical Bejo Zaden BV
Publication of WO2025140990A1 publication Critical patent/WO2025140990A1/fr
Pending legal-status Critical Current
Anticipated expiration legal-status Critical

Links

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
    • A01H5/00—Angiosperms, i.e. flowering plants, characterised by their plant parts; Angiosperms characterised otherwise than by their botanic taxonomy
    • A01H5/12—Leaves
    • 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
    • A01H1/12—Processes for modifying agronomic input traits, e.g. crop yield
    • A01H1/122—Processes for modifying agronomic input traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance
    • A01H1/1245—Processes for modifying agronomic input traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for biotic stress resistance, e.g. pathogen, pest or disease resistance
    • A01H1/1255—Processes for modifying agronomic input traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for biotic stress resistance, e.g. pathogen, pest or disease resistance for fungal resistance
    • 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
    • A01H6/00—Angiosperms, i.e. flowering plants, characterised by their botanic taxonomy
    • A01H6/02—Amaranthaceae or Chenopodiaceae, e.g. beet or spinach
    • A01H6/028—Spinacia oleracea [spinach]
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/415—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from plants
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
    • C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
    • C12N15/8271—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance
    • C12N15/8279—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for biotic stress resistance, pathogen resistance, disease resistance
    • C12N15/8282—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for biotic stress resistance, pathogen resistance, disease resistance for fungal resistance

Definitions

  • the present invention relates to genes, coding sequences and/or proteins providing resistance against Peronospora farinosa and especially to genes, coding sequences and/or proteins providing resistance against Peronospora farinosa in a spinach plant.
  • the present invention further relates to spinach plants comprising the present resistance genes, coding sequences and/or proteins and to methods for providing and/or identifying Peronospora farinosa resistant spinach plants using the present genes, coding sequences and/or proteins.
  • Spinach is commercially grown worldwide for its attractive and nutritious leaves. Spinach (Spinacia oleracea or S.
  • oleracea is a member of the Amaranthaceae family, subfamily Chenopodioideae. Other well-known family members include quinoa and beet. The latter is a cultivated plant of major importance for agriculture with sugar beet, red beet, and Swiss chard as examples.
  • nutritional value while providing only relatively small amounts of calories (23 for 100 grams of cooked spinach), spinach is a rich source of vitamins A, B2 (or folate), B6, C, E and K and, additionally, magnesium, manganese, calcium, potassium, iron, and dietary fibre.
  • Spinach flowering is induced by (long) day lengths and under optimal conditions can reach even up to 4 generations in a year with a life cycle from seed to new harvest completed within 3 months.
  • a bottleneck can be caused by seed dormancy.
  • Spinach is a wind pollinator and its pollen can reach far.
  • a spinach line is considered male if it converts from female or mixed flowering to (all) male flowering within a week.
  • Female spinach lines stay so for at least three weeks without producing any pollen.
  • Hybrids of spinach can be produced making use of plants which have a female flowering phase and plants which have a male flowering phase as pollinator. Before the female spinach plants develop male flowers, all female flowers are fertilized by the male spinach plant. The setting of seeds occurs rapidly within 3 days and after that the ripening of the seed takes approximately a month. Under optimal conditions commercial elite spinach lines are grown and harvested within 25 days. Breeding resulted in spinach plants which are rapid growing without premature flowering.
  • This type is used for industry (canned or frozen spinach, as well as processed food and baby food);
  • - Semi savoy is an intermediate type of spinach with a comparable texture as the savoy type but easy to clean as the smooth type of spinach. It is cultivated both for fresh market and industry.
  • An oriental type which is heat tolerant, has long petioles, pointed leaves with several side lobes and as plant has an upright growth.
  • Most spinach is produced at high plant densities for fresh market production creating an ideal environment for disease development.
  • organic vegetables i.e., vegetables that are obtained without the use of pesticides, fungicides, insecticides and without chemical treatment of the seeds. The challenge here is that such organic vegetable production conditions promote the development of plant diseases.
  • the present invention relates to a genomic sequence according to SEQ ID No.1; nucleotide sequence SEQ ID Nos.2; or amino acid sequence SEQ ID Nos.3. Additionally, the present invention relates to spinach plants comprising a resistance gene as defined above; or to Peronospora farinosa resistant spinach plants comprising a genomic sequence according to SEQ ID No.1; nucleotide sequence SEQ ID Nos.2; or amino acid sequence SEQ ID Nos.3. According to the present invention, the present resistance gene can be homozygous or heterozygous present in a Peronospora farinosa resistant spinach plant.
  • the spinach plants as defined above can be additionally resistant to Stempyllium and/or CMV.
  • a spinach plant (Spinacia oleracea) was identified with resistance to the pathogen Peronospora farinosa.
  • the selected plant was sequenced with Illumina sequencing.
  • the sequencing data has been mapped against the S. oleracea reference genome and de novo genome assemblies were performed.
  • Previously performed marker analysis resulted in the identification of a QTL.
  • Within the span of the QTL a highly interesting gene coding for a putative resistance protein was observed.
  • Comparative genomics between the S. oleracea reference genome and the de novo sequenced plant resulted in the identification of the resistance providing gene.
  • the genomic sequence of the identified gene is SEQ ID No.1. Subsequently, experiments were performed to obtain the cDNA sequence of the resistance gene.
  • a PCR was performed with a TS-specific oligo (CATTGCAAGCAGTGGTATCAAC) and a gene specific primer (CGTCGAGAGGAGATTGGTCA).
  • a gene specific primer CGTCGAGAGGAGATTGGTCA.
  • 3’ RACE cDNA was generated with an anchored oligo-d(T)23VN primer and M-MULV reverse transcriptase (ProtoScript First Strand cDNA Synthesis Kit (NEB # E6300L)) .
  • a gene specific forward primer (GTGTGTCCAAGAAAGCCCTG) and oligo- d(T)23VN ) primer were used to obtain a PCR product.
  • gRNAs guide RNAs
  • the mutation(s) introduced can be either insertion, deletion or another modification.
  • the CRISPR machinery, gRNA and Cas9 proteins can be delivered in spinach plant cells, via either PEG 10 mediated transfection of ribonucleoproteins (RNP), or Agrobacterium-mediated gene transfer. Edited cells can be regenerated to obtain viable plants. Plants that contain a mutation in the resistance providing gene according to the invention can be selfed to obtain spinach plants where the defective gene is present homozygous.

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Chemical & Material Sciences (AREA)
  • Botany (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Developmental Biology & Embryology (AREA)
  • General Health & Medical Sciences (AREA)
  • Environmental Sciences (AREA)
  • Molecular Biology (AREA)
  • Zoology (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • Physiology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biomedical Technology (AREA)
  • Biotechnology (AREA)
  • General Engineering & Computer Science (AREA)
  • Wood Science & Technology (AREA)
  • Physics & Mathematics (AREA)
  • Microbiology (AREA)
  • Plant Pathology (AREA)
  • Cell Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Medicinal Chemistry (AREA)
  • Natural Medicines & Medicinal Plants (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

La présente invention concerne des gènes, des séquences codantes et/ou des protéines conférant une résistance contre Peronospora farinosa et notamment des gènes, des séquences codantes et/ou des protéines conférant une résistance contre Peronospora farinosa dans un plant d'épinard. La présente invention concerne en outre des plants d'épinard comprenant les présents gènes de résistance, séquences codantes et/ou protéines et des procédés de fourniture et/ou d'identification de plants d'épinard résistants à Peronospora farinosa utilisant les présents gènes, séquences codantes et/ou protéines. Plus particulièrement, la présente invention concerne un gène de résistance capable de conférer une résistance à Peronospora farinosa dans l'épinard, la résistance étant comprise dans la séquence génomique de SEQ ID No 1, et des plants d'épinard comprenant le présent gène de résistance.
PCT/EP2024/087997 2023-12-29 2024-12-20 Gène conférant une résistance contre peronospora farinosa dans l'épinard Pending WO2025140990A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
EPPCT/EP2023/087993 2023-12-29
PCT/EP2023/087993 WO2025140780A1 (fr) 2023-12-29 2023-12-29 Gène conférant une résistance contre peronospora farinosa à l'épinard

Publications (1)

Publication Number Publication Date
WO2025140990A1 true WO2025140990A1 (fr) 2025-07-03

Family

ID=89573878

Family Applications (2)

Application Number Title Priority Date Filing Date
PCT/EP2023/087993 Pending WO2025140780A1 (fr) 2023-12-29 2023-12-29 Gène conférant une résistance contre peronospora farinosa à l'épinard
PCT/EP2024/087997 Pending WO2025140990A1 (fr) 2023-12-29 2024-12-20 Gène conférant une résistance contre peronospora farinosa dans l'épinard

Family Applications Before (1)

Application Number Title Priority Date Filing Date
PCT/EP2023/087993 Pending WO2025140780A1 (fr) 2023-12-29 2023-12-29 Gène conférant une résistance contre peronospora farinosa à l'épinard

Country Status (1)

Country Link
WO (2) WO2025140780A1 (fr)

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2018059651A1 (fr) 2016-09-30 2018-04-05 Rijk Zwaan Zaadteelt En Zaadhandel B.V. Procédé destiné à modifier le profil de résistance de spinacia oleracea au mildiou
WO2022008296A1 (fr) * 2020-07-10 2022-01-13 Enza Zaden Beheer B.V. Plant d'épinard résistant au mildiou et nouveau gène de résistance
WO2022111797A1 (fr) * 2020-11-24 2022-06-02 Bejo Zaden B.V. Plants d'épinards résistants à peronospora farinosa et à stemphylium vesicarium

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2018059651A1 (fr) 2016-09-30 2018-04-05 Rijk Zwaan Zaadteelt En Zaadhandel B.V. Procédé destiné à modifier le profil de résistance de spinacia oleracea au mildiou
WO2022008296A1 (fr) * 2020-07-10 2022-01-13 Enza Zaden Beheer B.V. Plant d'épinard résistant au mildiou et nouveau gène de résistance
WO2022111797A1 (fr) * 2020-11-24 2022-06-02 Bejo Zaden B.V. Plants d'épinards résistants à peronospora farinosa et à stemphylium vesicarium

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
ANONYMOUS: "Our Spinach varieties Enza Zaden resistant to new races of downy mildew | Enza Zaden", 28 April 2021 (2021-04-28), pages 1 - 2, XP093177848, Retrieved from the Internet <URL:https://www.enzazaden.com/news-and-events/news/2021/spinach-varieties-enza-zaden-resistant-to-new-races-of-downy-mildew> *
SAKATA VEGETABLES EUROPE S A S: "Spinacia oleracea L. ; Peridot ; spw20002", COMMUNITY PLANT VARIETY OFFICE, CPVO, 3 BOULEVARD MARÉCHAL FOCH CS 10121 49101 ANGERS CEDEX 2 - FRANCE, 25 May 2022 (2022-05-25), XP090012934 *

Also Published As

Publication number Publication date
WO2025140780A1 (fr) 2025-07-03

Similar Documents

Publication Publication Date Title
JP2023153180A (ja) トバモウイルスであるトマト褐色縮葉フルーツウイルス（ｔｂｒｆｖ）に対するソヌラム・リコペルシカム植物における耐性
Dida et al. Finger millet
Zhebentyayeva et al. Apricot
US20250084430A1 (en) Downy Mildew Resistant Spinach Plant
AU2017239475A1 (en) Hybrid seed potato breeding
Sarkar et al. Grass Pea: remodeling an ancient insurance crop for climate resilience
JP2022538791A (ja) トバモウイルスであるトマト褐色しわ果実ウイルスに対するトマトの植物体の抵抗力
WO2022111797A1 (fr) Plants d&#39;épinards résistants à peronospora farinosa et à stemphylium vesicarium
Polidoros et al. Lentil gene pool for breeding
US20250243505A1 (en) Genes From Spinacia Tetrandra Encoding a Protein Providing Resistance Against Peronospora Farinosa and Spinach Plants Comprising These Genes
IL303102A (en) Tomato plants resistant to TOMMV, TOMV, TMV, TOBRFV and suitable resistant genes
CN110891416B (zh) 腐霉属抗性的遗传基础
WO2025140990A1 (fr) Gène conférant une résistance contre peronospora farinosa dans l&#39;épinard
Boukar et al. Strategies in cowpea breeding
Zong et al. Genomic designing for climate-smart pea
US9580726B2 (en) Tomato plants comprising transgenic event Bs2-X5
US11997964B2 (en) Carrot variety NUN 85938 CAC
US11653605B2 (en) Hybrid carrot variety NUN 89733 CAC
Patto et al. Grass pea
Ijaz et al. 1 Conventional and Biotechnological Interventions in Summer Fodders
US10779488B2 (en) Carrot variety NUN 85935 CAC
US10779493B2 (en) Carrot variety NUN 85936 CAC
Paula BIOTECHNOLOGIES AND THEIR USE IN WHEAT RESEARCH IN ROMANIA
Salamini et al. Conventional and molecular plant breeding
US20250331489A1 (en) Bean plant named hmc010522

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 24836763

Country of ref document: EP

Kind code of ref document: A1