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CN-122012525-A - Resistance genes against plexiform root diseases

CN122012525ACN 122012525 ACN122012525 ACN 122012525ACN-122012525-A

Abstract

The present invention provides a novel nucleic acid molecule encoding a polypeptide which confers resistance to pathogens, in particular the "betanecrosis yellow vein virus" (BNYVV), in plants, in particular in Beta plants expressing the polypeptide, and also provides preferred nucleic acid molecules encoding the RZ-3 gene of Beta maritima, as well as derivatives and homologues thereof. Additional aspects of the invention include vectors, transgenic plant cells, transgenic plants and methods of making the same, and methods of identifying nucleic acid molecules that confer resistance.

Inventors

  • O. Terjek
  • D. Borchard
  • W. Mechelk
  • J.C. Lane

Assignees

  • KWS种子欧洲股份公司

Dates

Publication Date
20260512
Application Date
20140606
Priority Date
20130617

Claims (14)

  1. 1. A nucleic acid molecule encoding a polypeptide capable of conferring resistance to a pathogen in a plant expressing said polypeptide, characterized in that said nucleic acid molecule comprises a nucleotide sequence selected from the group consisting of: a) A nucleotide sequence encoding a polypeptide having the amino acid sequence of SEQ ID NO. 2 or SEQ ID NO. 3; b) A nucleotide sequence comprising the coding sequence of the DNA sequence of SEQ ID NO. 1; c) A nucleotide sequence which hybridizes under stringent conditions to the complement of the nucleotide sequence of a) or b); d) A nucleotide sequence encoding a polypeptide derived from the polypeptide encoded by the nucleotide sequence of a) or b) by one or more amino acid substitutions, deletions and/or additions in the amino acid sequence encoded by the nucleotide sequence of a) or b); e) A nucleotide sequence encoding a polypeptide having an amino acid sequence that is at least 60% identical to the amino acid sequence encoded by the nucleotide sequence of a) or b); f) Nucleotide sequence encoding at least one nucleotide binding domain (NBS) corresponding to amino acid positions 168-227 of SEQ ID NO. 2 or amino acid positions 182-241 of SEQ ID NO. 3, at least one leucine rich domain (LRR) corresponding to amino acid positions 591-613 of SEQ ID NO. 2 or amino acid positions 605-627 of SEQ ID NO. 3, and/or at least one internal repeat domain (IR) corresponding to amino acid positions 1013-1072 of SEQ ID NO. 2 or amino acid positions 1027-1086 of SEQ ID NO. 3.
  2. 2. A vector comprising the nucleic acid molecule of claim 1.
  3. 3. A host cell comprising the nucleic acid molecule of claim 1 or the vector of claim 2.
  4. 4. A polypeptide which confers resistance to a pathogen in a plant expressing the polypeptide and which is encoded by the nucleic acid molecule of claim 1.
  5. 5. A transgenic plant cell comprising the nucleic acid molecule of claim 1 as a transgene, or comprising the vector of claim 2.
  6. 6. A transgenic plant or part thereof comprising the plant cell of claim 5.
  7. 7. Seed of the plant of claim 6, wherein the seed comprises the nucleic acid molecule of claim 1 as transgene.
  8. 8. A method for producing a transgenic plant cell, characterized in that the method comprises the step of introducing the nucleic acid molecule of claim 1 or the vector of claim 2 into a plant cell.
  9. 9. A method for producing a transgenic plant, characterized in that the method comprises the steps of: a) Introducing the nucleic acid molecule of claim 1 or the vector of claim 2 into a plant cell, and B) Regenerating a transgenic plant from the transgenic plant cell of step a).
  10. 10. A promoter regulatory sequence which controls the expression of a gene comprising a nucleic acid molecule of the invention, characterized in that the regulatory sequence is capable of conferring or regulating the expression of a heterologous DNA sequence upon infection by a pathogen and the regulatory sequence comprises a nucleic acid molecule having the nucleotide sequence of nucleotides 1-1403 of SEQ ID No. 1.
  11. 11. A recombinant DNA molecule comprising the regulatory sequence of claim 10.
  12. 12. A host cell transformed with the regulatory sequence of claim 10 or with the recombinant DNA molecule of claim 11.
  13. 13. A transgenic plant, plant tissue or plant cell comprising as transgene the regulatory sequence of claim 10 or the recombinant DNA molecule of claim 11.
  14. 14. A method for identifying a nucleic acid molecule encoding a protein capable of conferring resistance to a pathogen BNYVV in a beta plant expressing the protein, characterized in that the method comprises the steps of: i detecting the absence of an insert in the coding nucleotide sequence of the nucleic acid molecule of claim 1, or Ii detecting a polymorphism in at least one of the coding nucleotide sequences of the nucleic acid molecule according to claim 1 according to figures 1, 2 and/or 3 using a molecular marker identifying said polymorphism.

Description

Resistance genes against plexiform root diseases The application is a divisional application of an application patent application with the application date of 2014, 6, 201480045665.2 and the application name of 'resistance gene against arbuscular diseases'. Technical Field The present invention relates to nucleic acid molecules encoding polypeptides which may confer (convey) resistance to pathogens, in particular "betanecrosis yellow vein virus" (Beet Necrotic Yellow Vein Virus, BNYVV), in plants, particularly in betalains expressing the polypeptides. The invention also relates to polypeptides which confer resistance to pathogens in plants, particularly BNYVV in beta plants expressing the polypeptides, and which are encoded by the nucleic acid molecules of the invention. The invention also relates to transgenic plants, plant cells, plant organs, plant tissues, plant parts or seeds of plants comprising the nucleic acid molecules or parts thereof, and to methods for producing transgenic plants or plant cells of this type. The invention also includes methods of detecting the resistance conferring nucleic acid molecules, and methods of screening plants or plant cells having the resistance conferring nucleic acid molecules. Background In terms of profit, arbuscular disease (Rhizomania) is the most serious beet disease worldwide, possibly resulting in 50% or more of revenue loss. The disease, also known as "mad root disease (root madness)", is caused by the "beet necrotic yellow virus" (Beet Necrotic Yellow Vein Virus, BNYVV) and is transmitted by the soil borne protozoan beet polymyxa (Polymyxa betae). BNYVV infection shows its increased proliferation in fine roots and secondary roots, and the formation of greatly reduced roots with reduced sugar content. Infected plants exhibit reduced water uptake and are therefore more susceptible to drought stress. When the infection propagates to the whole plant, it causes yellowing of the veins, necrotic lesions, and yellow spots on the leaves. Since, like other viral diseases, curative combating of such diseases is not possible, damage can only be prevented by breeding resistant varieties. Three major genes currently being tested for plexiform root disease are RZ-1 (also known as "Holly"), RZ-2 and RZ-3. Furthermore, more Cong Gen disease resistance genes are described in the literature, but these are of less importance. Currently, the resistance gene RZ-1 has been incorporated into most breeding lines (seed parent and/or pollinated parent components). However, resistance conferred by RZ-1 was found to be insufficient in severely affected areas or areas with multiple BNYVV pathotypes (e.g., sohi & Maleki, 2004). For this reason, it has been proposed in the past to use RZ-1 in combination with, for example, RZ-2 or RZ-3. RZ-2 and RZ-3 originate from beet (Beta vulgarissubsp. Maritime) sources (WB 42, WB 41) and are genetically localized to the same region on chromosome 3 in the beet genome, while RZ-1 is likewise localized to chromosome 3, but in the south of RZ-2 and RZ-3. Scholten et al (1999) determined the distance of 20-25 cM between RZ-1 and RZ-2, the major RZ genes. Gidner et al (2005) found a shorter distance of 5 cM between RZ-1 and RZ-2 and did not conclude that RZ-2 and RZ-3 were located at the same locus. SCHMIDLIN et al (2008) found differentially induced genes by expression analysis in infected beets, however these genes did not correspond to RZ-2 or RZ-3. In Larson et al (2008) studies, certain BNYVV-induced proteins were detected in beets by MALDI-TOF-MS, however scientists were not able to identify the proteins encoded by RZ-1, RZ-2 or RZ-3. Furthermore, the sequence region, especially in the vicinity of the resistance gene, is repetitive, which makes the development of diagnostic markers particularly difficult. Until now, high resolution marker profiles and proven candidate genes were not publicly available for specific Cong Gen disease resistance genes. In addition, the functional background, i.e., genetic structure, of these resistance genes has not been completely understood before. For sustainable planting of anti-clump root disease, which aims to offset the risk of breaking resistant BNYVV isolates, it is necessary to constantly identify new resistance genes and integrate these genes into the gene pool of crop plants (e.g. sugar beet). Disclosure of Invention The present invention has been developed based on the above prior art, wherein it is an object of the present invention to provide nucleic acid molecules and/or polypeptides which can confer Cong Gen disease resistance in plants. It is a further object to provide transgenic Cong Gen disease resistant plants, and methods of producing the same. It is a further object of the present invention to provide methods of using and developing molecular markers that allow for the efficient planting of anti Cong Gen disease and the development of new resistant plant lines. Embodiments of the inventio