Peronospora resistance in spinacia oleracea

Abstract

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 . 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%, 99.2%, 99.5%, 99.8%, or 100% sequence identity to SEQ ID NO: 10. 
     
     
         2 . The allele of  claim 1  wherein the genomic DNA sequence of the LRR domain in order of increased preference has at least 95%, 96%, 97%, 98%, 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, or 100% sequence identity to SEQ ID NO: 9. 
     
     
         3 . The allele of  claim 1 , wherein 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. 
     
     
         4 . A spinach plant comprising the allele of  claim 1 , of which a representative sample of seed capable of growing into a plant comprising said allele was deposited with the NCIMB under accession number NCIMB 43554. 
     
     
         5 . The spinach plant of  claim 4 , wherein the plant is an agronomically elite plant. 
     
     
         6 . The spinach plant of  claim 5 , wherein the agronomically elite plant is a hybrid variety or an inbred line. 
     
     
         7 . The spinach plant of  claim 6 , further comprising a genetic determinant resulting in resistance against  Peronospora farinosa  f. sp.  spinacea  races Pfs:1 to Pfs:17. 
     
     
         8 . A propagation material capable of developing into and/or being derived from the spinach plant of  claim 4 , wherein the propagation material comprises the allele and wherein the propagation material is selected from a group consisting of a microspore, a pollen, an ovary, an ovule, an embryo, an embryo sac, an egg cell, a cutting, a root, a root tip, a hypocotyl, a cotyledon, a stem, a leaf, a flower, an anther, a seed, a meristematic cell, a protoplast, a cell, or a tissue culture thereof. 
     
     
         9 . A cell of a spinach plant, which cell comprises the allele of  claim 1 . 
     
     
         10 . A method of producing a hybrid spinach seed comprising crossing a first parent spinach plant with a second parent spinach plant and harvesting the resultant hybrid spinach seed, wherein said first parent spinach plant comprises the allele of  claim 1 . 
     
     
         11 . The method of  claim 10 , wherein the first and/or second parent is a plant of an inbred line. 
     
     
         12 . A hybrid spinach plant grown from the seed produced by the method of  claim 10  or  claim 11 . 
     
     
         13 . A method for identifying a spinach plant carrying the allele of  claim 1 , 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 sequence has in order of increased preference 95%, 96%, 97%, 98%, 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, or 100% sequence identity to SEQ ID NO: 9. 
     
     
         14 . The method of  claim 13 , wherein the LRR domain is determined by using 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. 
     
     
         15 . The method of  claim 13 , wherein the LRR domain is determined by using 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. 
     
     
         16 . A primer pair comprising a forward primer which is a nucleic acid molecule having the sequence of SEQ ID NO: 4 and a reverse primer which is a nucleic acid molecule having the sequence of SEQ ID NO: 5. 
     
     
         17 . A method for producing a spinach plant showing resistance to  Peronospora farinosa  f. sp.  spinaciae  comprising:
 (a) crossing a plant comprising the allele 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 plant that comprises said allele.   
     
     
         18 . The method of  claim 17 , wherein the selection of a plant comprising the allele 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 sequence has in order of increased preference 95%, 96%, 97%, 98%, 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, or 100% sequence identity to SEQ ID NO: 9.