Peronospora resistance in spinacia oleracea

Abstract

The present invention relates to an allele designated alpha-WOLF 15 which 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:13) at its N-terminus; and b) the motif “KWMCLR” (SEQ ID NO:14; and wherein the LRR domain of the protein has in order of increased preference at least 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% sequence similarity to SEQ ID NO:10. The allele when homozygously present in a spinach plant confers complete resistance to Peronospora farinosa f. sp. spinaciae races Pfs:1, Pfs:2, Pfs:3, Pfs:4 and Pfs:5, Pfs:6, Pfs:8, Pfs:9, Pfs:11, Pfs:12, Pfs:13, Pfs:14, Pfs:15 and isolates UA1014, and confers intermediate resistance to pfs:10, and does not confer resistance to Pfs:7 and Pfs:16.

What is claimed is: 
     
         1 . An allele designated alpha-WOLF 15 which confers resistance to at least one  Peronospora farinosa  f. Sp.  spinaciae  race when present in a spinach plant, 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:13) at its N-terminus; and b) the motif “KWMCLR” (SEQ ID NO:14); and wherein the LRR domain of the protein has in order of increased preference at least 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% sequence similarity to SEQ ID NO:10. 
     
     
         2 . The allele of  claim 1 , wherein the allele when homozygously present in a spinach plant confers complete resistance to  Peronospora farinosa  f. sp.  spinaciae  races Pfs:1, Pfs:2, Pfs:3, Pfs:4 and Pfs: 5, Pfs:6, Pfs:8, Pfs:9, Pfs:11, Pfs:12, Pfs:13, Pfs:14, Pfs:15, Pfs:16 and isolates UA1014 and US1508, and confers intermediate resistance to Pfs:10, and does not confer resistance to Pfs:7. 
     
     
         3 . The allele of  claim 1 , wherein the allele has a genomic nucleotide sequence which in order of increased preference has at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% sequence similarity to SEQ ID NO:1. 
     
     
         4 . The allele of  claim 1 , wherein the allele has a coding sequence which in order of increased preference has at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% sequence similarity to SEQ ID NO:2. 
     
     
         5 . The allele of  claim 1 , wherein the allele has a coding sequence which in order of increased preference has at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% sequence similarity to SEQ ID NO:3. 
     
     
         6 . The allele of  claim 1 , wherein the allele encodes a protein having an amino acid sequence which in order of increased preference has at least 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% sequence similarity to SEQ ID NO:4. 
     
     
         7 . The allele of  claim 1 , wherein the allele encodes for a protein having an amino acid sequence which in order of increased preference has at least 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% sequence similarity to SEQ ID NO:5. 
     
     
         8 . 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 as claimed in  claim 1 . 
     
     
         9 . The method of  claim 8 , wherein the first and/or second parent is a plant of an inbred line. 
     
     
         10 . A hybrid spinach plant grown from the seed produced by the method of  claim 8 . 
     
     
         11 . A method for identifying or selecting a spinach plant carrying the allele as claimed in  claim 1 , comprising determining the presence of a genomic nucleotide sequence or a part thereof in the genome of a plant, wherein said sequence has in order of increased preference 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% sequence similarity to SEQ ID NO:1. 
     
     
         12 . A method for identifying or selecting a spinach plant carrying the allele as claimed in  claim 1 , comprising determining the presence of a genomic nucleotide sequence or a part thereof in the genome of a plant, wherein said sequence has in order of increased preference 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% sequence similarity to SEQ ID NO:2. 
     
     
         13 . A method for identifying or selecting a spinach plant carrying the allele as claimed in  claim 1 , comprising determining the presence of a genomic nucleotide sequence or a part thereof in the genome of a plant, wherein said sequence has in order of increased preference 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% sequence similarity to SEQ ID NO:3. 
     
     
         14 . The method as claimed in any of the  claims 11  to  13 , comprising determining the presence of the LRR domain as defined in  claim 1 . 
     
     
         15 . The method of  claim 14 , 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:6. 
     
     
         16 . The method of  claim 14 , 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:7. 
     
     
         17 . A primer pair comprising a forward primer which is a nucleic acid molecule having the sequence of SEQ ID NO:6 and a reverse primer which is a nucleic acid molecule having the sequence of SEQ ID NO:7. 
     
     
         18 . A method for producing a spinach plant showing resistance to  Peronospora farinosa  f. sp.  spinaciae  comprising: (a) crossing a plant comprising the allele as claimed in  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 as claimed in  claim 1 . 
     
     
         19 . The method of  claim 18 , wherein the selection of a plant comprising the allele comprises determining the presence of the allele according the method as claimed in any one of the  claims 11  to  13 .