EP2732039A1 - Procédé d'amélioration de la résistance des plantes aux virus - Google Patents
Procédé d'amélioration de la résistance des plantes aux virusInfo
- Publication number
- EP2732039A1 EP2732039A1 EP12738426.1A EP12738426A EP2732039A1 EP 2732039 A1 EP2732039 A1 EP 2732039A1 EP 12738426 A EP12738426 A EP 12738426A EP 2732039 A1 EP2732039 A1 EP 2732039A1
- Authority
- EP
- European Patent Office
- Prior art keywords
- plant
- protein
- csn5
- gene
- genome
- 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- Withdrawn
Links
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Classifications
-
- 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
- C12N15/8283—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 virus resistance
-
- 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
Definitions
- this complex In plants, this complex is described as having for its principal biological function a déddylation activity. This activity is mediated by complex subunit 5 (CSN5) (Schwechheimer et al., 2010). This CSN5 protein is highly conserved in all higher eukaryotes.
- the defective mutants for the gene encoding the CSN5 protein are preferably, but not exclusively, homozygous.
- the mutation of the The gene encoding the CSN5 protein is introduced into a gene exon. The invention does not exclude the fact that the mutation is located in an intron of the gene, insofar as this mutation affects the activity of the CSN5 protein encoded by this gene, in the sense recommended by the invention.
- Figures 1a and 1b are diagrams illustrating, for the mutants of Arabidopsis thaliana csn5a-1, csn5a-2, MIF20E3, MIF20E4, MIF20E6, MIF20E7 and for the wild line, the expression of a CSN5 gene measured by Quantitative RT-PCR, CSN5A for Figure 1a and CSN5B for Figure 1b.
- the PPV-R isolate exists as an infectious clone named pICPPV.
- This infectious clone was modified to insert a gene encoding green fluorescent protein (GFP), to give the infectious clone pICPPVnk-GFP (Fernandez-Fernandez et al., 2001).
- GFP green fluorescent protein
- TuMV is detected by symptomatology and RT-PCR.
- ELISA enzymatic immunoabsorption test
- the pair of P1 / P2 primers specific for PPV detection amplifies a 243 bp fragment of the N-terminal region of the virus capsid protein. This is demonstrated by migration on a 2% agarose gel. Previously extracted RNA corresponding to the PPV-R isolate is used as a positive control of the PCR.
- Primer P1 Sequence SEQ ID NO: 1
- RNAs were extracted from leaf tissue (100 mg) using the RNeasy® Mini Kit (QIAGEN) according to the supplier's recommendations. Contamination due to the presence of genomic DNA (gDNA) is eliminated by a Turbo® DNAse treatment according to the supplier's protocol (Ambion). The absence of contamination with gDNA is then verified by PCR using the primer pair specific for the following ACTINE gene: Actin25 primer: SEQ ID NO: 9
- These seeds are harvested, decontaminated and kept for 2 days at 4 ° C in water containing kanamycin at 50 ⁇ 9. ⁇ 1.
- the seeds are then taken up in sterile lukewarm agarose at 0.05% (w / v) and spread on culture medium (MS medium pH 5.7, diluted to half, agar, glucose 5% (w / v) )) added with kanamycin at 50 ⁇ 9. ⁇ 1 .
- Kanamycin resistant plantlets, which overexpress MIF2 are transferred to soil.
- the cylindrical inclusion gene of the Turnip mosaic virus encodes a pathogenic determinant to the Brassica resistance gene TuRBOL Mol. Plant- Microb. Interact. 13, 1 102-1 108.
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Genetics & Genomics (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Zoology (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
- Biophysics (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Physics & Mathematics (AREA)
- Microbiology (AREA)
- Plant Pathology (AREA)
- Cell Biology (AREA)
- Virology (AREA)
- Botany (AREA)
- Gastroenterology & Hepatology (AREA)
- Medicinal Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
- Peptides Or Proteins (AREA)
Abstract
Description
Claims
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
FR1156258A FR2977896A1 (fr) | 2011-07-11 | 2011-07-11 | Procede d'amelioration de la resistance des plantes aux virus |
PCT/EP2012/063513 WO2013007728A1 (fr) | 2011-07-11 | 2012-07-10 | Procédé d'amélioration de la résistance des plantes aux virus |
Publications (1)
Publication Number | Publication Date |
---|---|
EP2732039A1 true EP2732039A1 (fr) | 2014-05-21 |
Family
ID=46579000
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
EP12738426.1A Withdrawn EP2732039A1 (fr) | 2011-07-11 | 2012-07-10 | Procédé d'amélioration de la résistance des plantes aux virus |
Country Status (4)
Country | Link |
---|---|
US (1) | US20140230089A1 (fr) |
EP (1) | EP2732039A1 (fr) |
FR (1) | FR2977896A1 (fr) |
WO (1) | WO2013007728A1 (fr) |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
GB201913060D0 (en) * | 2019-09-10 | 2019-10-23 | Innes John Centre | Methods of increasing biotic stress resistance in plants |
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2011
- 2011-07-11 FR FR1156258A patent/FR2977896A1/fr not_active Withdrawn
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2012
- 2012-07-10 EP EP12738426.1A patent/EP2732039A1/fr not_active Withdrawn
- 2012-07-10 US US14/131,529 patent/US20140230089A1/en not_active Abandoned
- 2012-07-10 WO PCT/EP2012/063513 patent/WO2013007728A1/fr active Application Filing
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US20140230089A1 (en) | 2014-08-14 |
FR2977896A1 (fr) | 2013-01-18 |
WO2013007728A1 (fr) | 2013-01-17 |
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