Compositions and methods for peronospora resistance in spinach

Abstract

The invention provides for spinach plants with broad-spectrum resistance to downy mildew disease and their progeny. Such plants may comprise unique combinations of alleles resulting in the broad-spectrum resistance to downy mildew. In certain aspects, compositions, including distinct polymorphic molecular markers, and methods for producing, using, identifying, selecting, and the like of plants or germplasm with resistance to downy mildew are provided.

1 . A  Spinacia oleracea  spinach plant of a cultivated variety comprising in its genome allele A, wherein said plant comprises a DNA sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:19, and SEQ ID NO:22, and comprises the DNA sequence of SEQ ID NO:5 or SEQ ID NO:10, and wherein a sample of seed comprising said allele A has been deposited under ATCC Accession No. PTA-120472. 
     
     
         2 . A  Spinacia oleracea  spinach plant of a cultivated variety comprising in its genome a heterozygous combination of alleles that confers broad-spectrum resistance to  Peronospora farinosa  f. sp.  Spinaciae  (Pfs), wherein the combination of alleles comprises two alleles selected from the group consisting of allele A, allele Vt, and allele C, and wherein a plant comprising allele A comprises a DNA sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:19, and SEQ ID NO:22, and comprises the DNA sequence of SEQ ID NO:5 or SEQ ID NO:10, wherein a plant comprising allele Vt comprises a DNA sequence selected from the group consisting of SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:11, SEQ ID NO:14, and SEQ ID NO:17 and comprises the DNA sequence of SEQ ID NO: 20 or SEQ ID NO: 23, and wherein a plant comprising allele C comprises a DNA sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:18, and SEQ ID NO:21 and comprises the DNA sequence of SEQ ID NO:4, SEQ ID NO:9, SEQ ID NO:12, and SEQ ID NO: 15, and wherein a sample of seed comprising said allele A, allele Vt, and allele C has been deposited under ATCC Accession No. PTA-120472, ATCC Accession No. PTA-12041, and ATCC Accession No. PTA-12486, respectively. 
     
     
         3 . The spinach plant of  claim 2 , wherein the broad-spectrum resistance comprises resistance to at least 10 races of  Peronospora farinosa  f. sp.  spinaciae  (Pfs). 
     
     
         4 . (canceled) 
     
     
         5 . The spinach plant of  claim 2 , wherein the combination of alleles comprises:
 (a) alleles A and C;   (b) alleles A and Vt; or   (c) alleles C and Vt.   
     
     
         6 . The spinach plant of  claim 3 , wherein the plant is resistant to:
 (a)  Peronospora farinosa  f. sp.  Spinaciae  races, 7, 8, 10, 11, 12, 14, and isolate UA4712;   (b)  Peronospora farinosa  f. sp.  Spinaciae  races, 7, 8, 10, 11, 12, 13, and 14; or   (c)  Peronospora farinosa  f. sp.  Spinaciae  races, 7, 8, 10, 11, 12, 13, and isolate UA4712.   
     
     
         7 .- 8 . (canceled) 
     
     
         9 . The spinach plant of  claim 1 , wherein said plant is an inbred. 
     
     
         10 . The spinach plant of  claim 1 , wherein said plant is a hybrid. 
     
     
         11 . The spinach plant of  claim 1 , comprising recessive resistance to  Peronospora farinosa  f. sp.  Spinaciae  races 7 and 13. 
     
     
         12 . A seed that produces the plant of  claim 2 . 
     
     
         13 . A plant part of the plant of  claim 2 , wherein the plant is a leaf, stem, root, flower or cell. 
     
     
         14 . A method of producing a seed of a spinach plant with broad spectrum resistance to  Peronospora farinosa  f. sp.  Spinaciae  comprising:
 (a) crossing a first spinach plant and a second spinach plant, wherein the first and second spinach plants collectively comprise at least two alleles selected from the group consisting of allele A, allele Vt, and allele C, wherein a plant comprising allele A comprises a DNA sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:19, and SEQ ID NO:22, and comprises the DNA sequence of SEQ ID NO:5 or SEQ ID NO:10, wherein a plant comprising allele Vt comprises a DNA sequence selected from the group consisting of SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:11, SEQ ID NO:14, and SEQ ID NO:17 and comprises the DNA sequence of SEQ ID NO: 20 or SEQ ID NO: 23, and wherein a plant comprising allele C comprises a DNA sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:18, and SEQ ID NO:21 and comprises the DNA sequence of SEQ ID NO:4, SEQ ID NO:9, SEQ ID NO:12, and SEQ ID NO: 15, and wherein a sample of seed comprising said allele A, allele Vt, and allele C has been deposited under ATCC Accession No. PTA-120472, ATCC Accession No. PTA-12041, and ATCC Accession No. PTA-12486, respectively;   (b) obtaining at least a first seed resulting from said crossing that comprises a heterozygous combination of said alleles that confers broad-spectrum resistance to  Peronospora farinosa  f. sp.  Spinaciae.      
     
     
         15 . The method of  claim 14 , wherein the method further comprises obtaining a population of seed resulting from said crossing that comprises a heterozygous combination of said alleles that confers broad-spectrum resistance to  Peronospora farinosa  f. sp.  Spinaciae.    
     
     
         16 . The method of  claim 14 , wherein:
 (a) the first spinach plant comprises allele A and the second spinach plant comprises allele C;   (b) the first spinach plant comprises allele A and the second spinach plant comprises allele Vt;   (c) the first spinach plant comprises allele C and the second spinach plant comprises allele Vt;   (d) the first spinach plant or the second spinach plant is homozygous for said allele A, allele Vt, or allele C; or   (e) the first spinach plant and the second spinach plant are homozygous for said two alleles selected from the group consisting of allele A, allele Vt, and allele C.   
     
     
         17 . A seed produced by the method of  claim 14 . 
     
     
         18 . The seed of  claim 17 , wherein:
 (a) the first allele is allele A and the second allele is allele C;   (b) the first allele is allele A and the second allele is allele Vt; or   (c) the first allele is allele C and the second allele is allele Vt.   
     
     
         19 . A DNA sequence selected from the group consisting of SEQ ID NOs:1-25. 
     
     
         20 . A method of introducing resistance to  Peronospora farinosa  f. sp.  Spinaciae  into a spinach plant comprising:
 (a) crossing a first spinach plant comprising in its genome a first allele selected from the group consisting of A, Vt, and C, with a second spinach plant, wherein a plant comprising allele A comprises a DNA sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:19, and SEQ ID NO:22, and comprises the DNA sequence of SEQ ID NO:5 or SEQ ID NO:10, wherein a plant comprising allele Vt comprises a DNA sequence selected from the group consisting of SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:11, SEQ ID NO:14, and SEQ ID NO:17 and comprises the DNA sequence of SEQ ID NO: 20 or SEQ ID NO: 23, and wherein a plant comprising allele C comprises a DNA sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:18, and SEQ ID NO:21 and comprises the DNA sequence of SEQ ID NO:4, SEQ ID NO:9, SEQ ID NO:12, and SEQ ID NO: 15, and wherein a sample of seed comprising said allele A, allele Vt, and allele C has been deposited under ATCC Accession No. PTA-120472, ATCC Accession No. PTA-12041, and ATCC Accession No. PTA-12486, respectively;   (b) selecting at least one progeny plant that comprises said first allele for resistance to  Peronospora farinosa  f. sp.  Spinaciae,  by detecting the presence in the genome of said progeny plant the DNA sequence of  claim 19 .   
     
     
         21 . The method of  claim 20 , wherein said crossing comprises producing a population of progeny plants. 
     
     
         22 . The method of  claim 20 , comprising screening the progeny plants for the presence of at least two nucleic acid sequences selected from the group consisting of SEQ ID NOs: 1-25. 
     
     
         23 . A plant produced by the method of  claim 20 . 
     
     
         24 . A method of identifying a plant comprising a desired allele conferring resistance to  Peronospora farinosa  f. sp.  Spinaciae,  the method comprising detecting in the genome of said plant the DNA sequence of  claim 19 . 
     
     
         25 . The method of  claim 24 , further defined as comprising detecting in the genome of said plant at least two polymorphic nucleic acid sequences selected from the group consisting of SEQ ID NOs:1-25, wherein the presence of the polymorphic nucleic acid sequences are indicative of the presence in the plant of at least two alleles conferring resistance to  Peronospora farinosa  f. sp.  Spinaciae  selected from of alleles A, Vt, and C, wherein a plant comprising allele A comprises a DNA sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:19, and SEQ ID NO:22, and comprises the DNA sequence of SEQ ID NO:5 or SEQ ID NO:10, wherein a plant comprising allele Vt comprises a DNA sequence selected from the group consisting of SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:11, SEQ ID NO:14, and SEQ ID NO:17 and comprises the DNA sequence of SEQ ID NO: 20 or SEQ ID NO: 23, and wherein a plant comprising allele C comprises a DNA sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:18, and SEQ ID NO:21 and comprises the DNA sequence of SEQ ID NO:4, SEQ ID NO:9, SEQ ID NO:12, and SEQ ID NO: 15, and wherein a sample of seed comprising said allele A, allele Vt, and allele C has been deposited under ATCC Accession No. PTA-120472, ATCC Accession No. PTA-12041, and ATCC Accession No. PTA-12486, respectively. 
     
     
         26 . A plant obtained by the method of  claim 24 . 
     
     
         27 . A spinach plant, cell or cell containing the plant part of  claim 2 .