Peronospora resistance in spinacia oleracea
The present invention relates to an allele designated alpha-WOLF 24 which confers resistance to at least one Peronospora farinosa f. sp. spinacea race, wherein the protein encoded by said allele is a CC-NBS-LRR protein that comprises in its amino acid sequence: a) the motif “MAEIGYSVC” (SEQ ID NO: 1) at its N-terminus; and b) the motif “KWMCLR” (SEQ ID NO: 2); and wherein the LRR domain of the protein has in order of increased preference at least 95%, 96%, 97%, 98%, 98.2%, 98.5%, 98.8%, 99%, 100% sequence similarity to SEQ ID NO: 10. The allele when present in a spinach plant confers complete resistance to at least Peronospora farinosa f. sp. spinacea race Pfs: 1, Pfs: 2, Pfs: 5, Pfs: 6, Pfs: 7, Pfs: 9, Pfs: 11, Pfs: 13, Pfs: 15 and Pfs: 17, and does not confer resistance to downy mildew race Pfs: 16.
1 . A method for identifying a spinach plant carrying an allele designated alpha-WOLF 24;
wherein the allele confers resistance to at least one Peronospora farinosa f. sp. spinaciae race;
wherein the protein encoded by said allele is a CC-NBS-LRR protein that comprises in its amino acid sequence a) the motif “MAEIGYSVC” (SEQ ID NO: 1) at its N-terminus; and b) the motif “KWMCLR” (SEQ ID NO: 2); and
wherein the method comprises determining the presence of the LRR domain by determining its genomic nucleotide sequence or a part thereof in the genome of a plant, wherein said genomic nucleotide sequence has the sequence of SEQ ID NO: 9, and selecting the plant comprising the LRR domain, thereby identifying the spinach plant carrying the allele designated alpha-WOLF 24.
2 . The method of claim 1 , wherein the LRR domain is determined with a primer pair to amplify the LRR domain, wherein the forward primer is a nucleic acid molecule having the sequence of SEQ ID NO: 4.
3 . The method of claim 1 , wherein the LRR domain is determined with a primer pair to amplify the LRR domain, wherein the reverse primer is a nucleic acid molecule having the sequence of SEQ ID NO: 5.
4 . A method for producing a spinach plant showing resistance to Peronospora farinosa f. sp. spinaciae comprising:
(a) crossing the spinach plant identified to carry the allele designated alpha-WOLF 24 by the method of claim 1 , with another plant;
(b) optionally performing one or more rounds of selfing and/or crossing;
(c) selecting after one or more rounds of selfing and/or crossing for a spinach plant comprising the allele designated alpha-WOLF 24;
wherein the selection of the spinach plant comprising the allele designated alpha-WOLF 24 comprises determining the presence of the allele comprising determining the presence of the LRR domain by determining its genomic nucleotide sequence or a part thereof in the genome of a plant, wherein said genomic sequence has the sequence of SEQ ID NO: 9.