Peronospora resistance in spinacia oleracea
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-NB S-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.
1. An agronomically elite spinach plant comprising an allele which confers resistance to at least one Peronospora farinosa f. sp. spinaciae race when present in a spinach plant and encodes a protein that has at least 99% sequence identity to a protein comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 4 and 5; wherein said protein comprises in its amino acid sequence:
a) SEQ ID NO: 13,
b) SEQ ID NO: 14,
and wherein the LRR domain of the protein has at least 99% sequence identity to SEQ ID NO: 10.
2. An agronomically elite spinach plant comprising an allele which when homozygously present in a spinach plant encodes a protein that confers complete resistance to at least Peronospora farinosa f. sp. spinaciae races Pfs:1, Pfs:2, Pfs:3, Pfs:4, Pfs:5, Pfs:6, Pfs:8, Pfs:9, Pfs:11, Pfs:12, Pfs:13, Pfs:14, Pfs:15, and isolate UA1014, and intermediate resistance to Pfs:10, and does not confer resistance to Pfs:7 and Pfs:16, wherein the allele has a genomic nucleotide sequence which is SEQ ID NO: 1.
3. The agronomically elite spinach plant of claim 1 , wherein the allele encodes a protein having an amino acid sequence comprising SEQ ID NO: 4.
4. The agronomically elite spinach plant of claim 1 , wherein the allele encodes a protein having an amino acid sequence comprising SEQ ID NO: 5.
5. A method of producing an F1 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 and/or said second parent spinach plant is the agronomically elite spinach plant as claimed in claim 1 .
6. The method of claim 5 , wherein the first and/or second parent is a plant of an inbred line.
7. An F1 hybrid spinach plant grown from the seed produced by the method of claim 5 , wherein the F1 hybrid plant carries the allele which confers resistance to at least one Peronospora farinosa f sp. spinaciae race when present in a spinach plant and encoding a CC-NB S-LRR protein that has at least 99% sequence identity to a protein comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 4 and SEQ ID NO: 5; wherein said protein comprises in its amino acid sequence:
a) SEQ ID NO: 13,
b) SEQ ID NO: 14,
and wherein the LRR domain of the protein has at least 99% sequence identity to SEQ ID NO: 10.
8. A method for producing a spinach plant showing resistance to Peronospora farinosa f sp. spinaciae comprising: (a) crossing the plant as claimed in claim 1 or 2 , with another spinach plant; (b) optionally performing one or more rounds of selfing and/or crossing; (c) optionally selecting after the crossing or the one or more rounds of selfing and/or crossing for a plant that comprises said allele.
9. The method of claim 8 , wherein the method includes performing the optional selection, and the selection of the plant comprising the allele expressing the protein comprises determining the presence of the allele according to a method comprising any one or more of:
determining the presence of a genomic nucleotide sequence in the genome of a plant, wherein said sequence has at least 99% sequence identity to SEQ ID NO: 1, or
determining the presence of a nucleotide sequence in a plant, wherein said sequence has at least 99% sequence identity to SEQ ID NO: 2, or
determining the presence of a nucleotide sequence in a plant, wherein said sequence has at least 99% sequence identity to SEQ ID NO: 3 or
determining the presence of a LRR domain as having at least 99% sequence identity to SEQ ID NO: 9.
10. A method for producing a spinach plant comprising: (a) crossing a spinach plant comprising a nucleotide sequence having at least 99% sequence identity to SEQ ID NO: 1 or having a nucleotide sequence encoding a protein having at least 99% sequence identity to SEQ ID NO: 4 or 5 with another spinach plant; (b) optionally performing one or more rounds of selfing and/or crossing; (c) optionally selecting after the crossing or the one or more rounds of selfing and/or crossing for a plant that comprises said allele.
11. The method of claim 10 , wherein the method includes performing the optional selection, and the selection of a plant comprising the allele expressing the protein comprises determining the presence of the allele according to a method comprising any one or more of:
determining the presence of a genomic nucleotide sequence in a plant, wherein said sequence has at least 99% sequence identity to SEQ ID NO: 1, or
determining the presence of a nucleotide sequence in a plant, wherein said sequence has at least 99% sequence identity to SEQ ID NO: 2, or
determining the presence of a nucleotide sequence in a plant, wherein said sequence has at least 99% sequence identity to SEQ ID NO: 3.
12. The method of claim 11 further comprising determining the presence of a LRR domain as having at least 99% sequence identity to SEQ ID NO: 10.
13. A method of producing an F1 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 and/or said second parent spinach plant comprises a nucleic acid encoding a protein having at least 99% sequence identity to SEQ ID NO: 4 or 5, or a nucleic acid having at least 99% sequence identity to SEQ ID NO: 1.
14. The plant of claim 1 , wherein the allele encodes a protein having at least 96% sequence identity to SEQ ID NO: 4 or 5.
15. The plant of claim 1 , wherein the allele encodes a protein having at least 97% sequence identity to SEQ ID NO: 4 or 5.
16. The plant of claim 1 , wherein the allele encodes a protein having at least 98% sequence identity to SEQ ID NO: 4 or 5.
17. The plant of claim 1 , wherein the allele encodes a protein having at least 99% sequence identity to SEQ ID NO: 4 or 5.
18. The plant of claim 1 , wherein the allele encodes a protein having at least 100% sequence identity to SEQ ID NO: 4 or 5.
19. The plant of claim 6 , wherein the allele encodes a protein having at least 96% sequence identity to SEQ ID NO: 4.
20. The plant of claim 6 , wherein the allele encodes a protein having at least 97% sequence identity to SEQ ID NO: 4.
21. The plant of claim 6 , wherein the allele encodes a protein having at least 98% sequence identity to SEQ ID NO: 4.
22. The plant of claim 6 , wherein the allele encodes a protein having at least 99% sequence identity to SEQ ID NO: 4.
23. The plant of claim 6 , wherein the allele encodes a protein having at least 100% sequence identity to SEQ ID NO: 4.
24. The plant of claim 4 , wherein the allele encodes a protein having at least 96% sequence identity to SEQ ID NO: 5.
25. The plant of claim 4 , wherein the allele encodes a protein having at least 97% sequence identity to SEQ ID NO: 5.
26. The plant of claim 4 , wherein the allele encodes a protein having at least 98% sequence identity to SEQ ID NO: 5.
27. The plant of claim 4 , wherein the allele encodes a protein having at least 99% sequence identity to SEQ ID NO: 5.
28. The plant of claim 4 , wherein the allele encodes a protein having at least 100% sequence identity to SEQ ID NO: 5.
29. The plant of claim 2 , wherein the allele has at least 96% sequence identity to SEQ ID NO: 1.
30. The plant of claim 2 , wherein the allele has at least 97% sequence identity to SEQ ID NO: 1.
31. The plant of claim 2 , wherein the allele has at least 98% sequence identity to SEQ ID NO: 1.
32. The plant of claim 2 , wherein the allele has at least 99% sequence identity to SEQ ID NO: 1.
33. The plant of claim 2 , wherein the allele has 100% sequence identity to SEQ ID NO: 1.
34. The method of claim 10 , wherein the allele has at least 96% sequence identity to SEQ ID NO: 1.
35. The method of claim 10 , wherein the allele has at least 97% sequence identity to SEQ ID NO: 1.
36. The method of claim 10 , wherein the allele has at least 98% sequence identity to SEQ ID NO: 1.
37. The method of claim 10 , wherein the allele has at least 99% sequence identity to SEQ ID NO: 1.
38. The method of claim 10 , wherein the allele has 100% sequence identity to SEQ ID NO: 1.
39. The method of claim 10 , wherein the allele encodes a protein having at least 96% sequence identity to SEQ ID NO: 4 or 5.
40. The method of claim 10 , wherein the allele encodes a protein having at least 97% sequence identity to SEQ ID NO: 4 or 5.
41. The method of claim 10 , wherein the allele encodes a protein having at least 98% sequence identity to SEQ ID NO: 4 or 5.
42. The method of claim 10 , wherein the allele encodes a protein having at least 99% sequence identity to SEQ ID NO: 4 or 5.
43. The method of claim 10 , wherein the allele encodes a protein having at least 100% sequence identity to SEQ ID NO: 4 or 5.
44. The method of claim 11 , wherein the determining the presence of the allele comprises determining the presence of a genomic nucleotide sequence or a part thereof in a genomic plant, wherein the sequence has at least 96% sequence identity to SEQ ID NO: 1, 2, or 3.
45. The method of claim 11 , wherein the determining the presence of the allele comprises determining the presence of a genomic nucleotide sequence or a part thereof in a genomic plant, wherein the sequence has at least 97% sequence identity to SEQ ID NO: 1, 2, or 3.
46. The method of claim 11 , wherein the determining the presence of the allele comprises determining the presence of a genomic nucleotide sequence or a part thereof in a genomic plant, wherein the sequence has at least 98% sequence identity to SEQ ID NO: 1, 2, or 3.
47. The method of claim 11 , wherein the determining the presence of the allele comprises determining the presence of a genomic nucleotide sequence or a part thereof in a genomic plant, wherein the sequence has at least 99% sequence identity to SEQ ID NO: 1, 2, or 3.
48. The method of claim 11 , wherein the determining the presence of the allele comprises determining the presence of a nucleotide sequence in a plant, wherein the sequence has 100% sequence identity to SEQ ID NO: 1, 2, or 3.
49. The method of claim 12 , wherein the determining the presence a LRR domain comprises determining the presence of a LRR domain having at least 96% sequence identity to SEQ ID NO: 10.
50. The method of claim 12 , wherein the determining the presence a LRR domain comprises determining the presence of a LRR domain having at least 97% sequence identity to SEQ ID NO: 10.
51. The method of claim 12 , wherein the determining the presence a LRR domain comprises determining the presence of a LRR domain having at least 98% sequence identity to SEQ ID NO: 10.
52. The method of claim 12 , wherein the determining the presence a LRR domain comprises determining the presence of a LRR domain having at least 99% sequence identity to SEQ ID NO: 10.
53. The method of claim 12 , wherein the determining the presence a LRR domain comprises determining the presence of a LRR domain having at least 100% sequence identity to SEQ ID NO: 10.