Novel class of proteins and uses thereof for plant resistance to various pathogenic agents
AssigneesInstitut National de le Recherche Agronomique (INRA), Centre National de la Recherche Scientifique (CNRS)
InventorsJocelyne Olivier, Laurent Deslandes, Yves Marco
Life of this patent
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Abstract
Disclosed is substantially pure DNA encoding an Arabidopsis thaliana Rps2 polypeptide; substantially pure Rps2 polypeptide; and methods of using such DNA to express the Rps2 polypeptide in plant cells and whole plants to provide, in transgenic plants, disease resistance to pathogens. Also disclosed are conserved regions characteristic of the RPS family and primers and probes for the identification and isolation of additional RPS disease-resistance genes.
Claims57
Nucleic acid containing at least 15 consecutive nucleotides of a nucleotide sequence coding for a protein of resistance of a plant to a pathogen, said protein comprising:
- a) an N-terminal portion containing at least one amino acid sequence rich in leucine and at least one nucleotide-binding site;
- and
- b) a C-terminal portion containing a DNA-binding domain, said binding domain comprising the amino acid sequence “WRKYGQK”, as well as a nucleic acid of complementary sequence.
Fusion polypeptide, characterized in that it comprises the N-terminal portion of the RRS1-S polypeptide fused with the C-terminal portion of the RRS1-R polypeptide.
Fusion polypeptide, characterized in that it comprises the N-terminal portion of the RRS1-R polypeptide fused with the C-terminal portion of the RRS1-S polypeptide.
Method for screening a candidate substance fixing to a RRS1-S or RRS1-R polypeptide, characterized in that it comprises the steps of:
- a) obtaining a first nucleic acid coding for a fusion protein comprising a part of the polypeptide of interest fused to the DNA-binding domain of a transcription factor such as Gal4;
- b) obtaining a second nucleic acid coding for a fusion protein comprising the candidate substance fused to the transcription domain of a transcription factor such as Gal4;
- c) producing a nucleic acid containing a nucleotide sequence coding for a detectable marker, placed under the control of a regulatory sequence recognized by a transcription factor such as Gal4;
- the nucleic acids a), b) and c) being inserted into appropriate vectors;
- and
- d) co-transfecting yeast cells simultaneously with said vectors;
- e) detecting the expression of the nucleotide sequence coding for the detectable marker.
Kit or pack for screening a candidate substance fixing to the RRS1-S or RRS1-R polypeptide, comprising:
- a) a first nucleic acid coding for a fusion protein comprising a part of the polypeptide of interest fused to the DNA-binding domain of a transcription factor such as Gal4;
- b) optionally, a second nucleic acid containing a nucleotide sequence coding for a detectable marker, placed under the control of a regulatory sequence recognized by the transcription factor such as Gal4;
- c) a third nucleic acid coding for a fusion protein comprising the candidate substance fused to the transcription domain of the transcription factor such as Gal4;
Dependencies are inferred from the claim text.