Methods and compositions for identifying downy mildew resistant cucumber plants

• US 8,809,622 B2
• Filed: 10/22/2010
• Issued: 08/19/2014
• Est. Priority Date: 10/22/2009
• Status: Active Grant

• Current Assignee: Seminis Vegetable Seeds Incorporated (Bayer AG)
• Original Assignee: Seminis Vegetable Seeds Incorporated (Bayer AG)
• Inventors: Caldwell, David, Chan, Eva, Joobeur, Tarek, King, Joseph, Reina, Antonio, Shetty, Nischit, de Vries, Jeroen
• Primary Examiner(s): Zhou, Shubo (Joe)
• Assistant Examiner(s): ROBINSON, KEITH O NEAL
• Application Number:
  US12/910,478
• Publication Number:
  US 20110126309A1
• Time in Patent Office: 1,397 Days
• Field of Search: None
• US Class Current: 800/267
• CPC Class Codes: A01H 1/045 using molecular markers
• CPC Class Codes: A01H 5/08 Fruits
• CPC Class Codes: A01H 6/346 Cucumis sativus[cucumber]
• CPC Class Codes: C12Q 1/6895 for plants, fungi or algae
• CPC Class Codes: C12Q 2600/13 Plant traits
• CPC Class Codes: C12Q 2600/156 Polymorphic or mutational markers
• CPC Class Codes: C12Q 2600/172 Haplotypes

Abstract

The present invention relates to methods for identifying cucumber lines having increased resistance to Downy Mildew, and identification of genetic markers linked to gene(s) conditioning such increased disease resistance. The present invention also relates to methods of breeding cucumber plants from lines having increased Downy Mildew resistance by marker-assisted selection, compositions including nucleic acid probes or primers which are useful for such marker assisted selection, and plants and plant parts produced by such methods.

Claims

1. A method of obtaining cucumber germplasm comprising the steps of:

a) assaying cucumber plants for the presence of at least a first genetic marker genetically linked to a QTL that confers resistance to Downy Mildew; and

b) selecting at least a first cucumber plant comprising the genetic marker and the QTL that confers resistance to Downy Mildew;

wherein the QTL maps to a position between the sequence represented by SNP marker NN0228457 and SNP marker NN0228148, which map to approximately 25.2 cM and 82.7 cM on the genetic map of the linkage group termed cucumber chromosome 5, respectively;

wherein the QTL maps to a position between the sequence represented by SNP marker NN0225012 and SNP marker NN0227587 which map to approximately 20.6 cM and 87.4 cM on the genetic map of the linkage group termed cucumber chromosome 4, respectively;

or wherein the QTL maps to a position between the sequence represented by SNP marker NN0223782 and SNP marker NN0226638 which map to approximately 22.5 cM and 88.4 cM on the genetic map of the linkage group termed cucumber chromosome 2, respectively.

2. The method of claim 1, wherein the QTL maps to a position between the sequence represented by SNP marker NN0227981 and SNP marker NN0226631, which map to approximately 35.7 cM and 55.5 cM on the genetic map of the linkage group termed cucumber chromosome 5, respectively;

or wherein the QTL maps to a position between the sequence represented by SNP marker NN0227981and SNP marker NN0247786, which map to approximately 35.7 cM and 71.4 cM on the genetic map of the linkage group termed cucumber chromosome 5, respectively.

3. The method of claim 1, wherein the QTL maps to a position between the sequence represented by SNP marker NN0228579 and SNP marker NN0224495 which map to approximately 25.8 cM and 82.0 cM on the genetic map of the linkage group termed cucumber chromosome 4, respectively;

wherein the QTL maps to a position between the sequence represented by SNP marker NN0225088and SNP marker NN0224041 which map to approximately 30.4 cM and 75.8 cM on the genetic map of the linkage group termed cucumber chromosome 4, respectively;

or wherein the QTL maps to a position between the sequence represented by SNP marker NN0228579 and SNP marker NN0224495 which map to approximately 54.5 cM and 82.0 cM on the genetic map of the linkage group termed cucumber chromosome 4,respectively.

4. The method of claim 1, wherein the QTL maps to a position between the sequence represented by SNP marker NN0246378 and SNP marker NN0246472 which map to approximately 14.6 cM and 38.4 cM on the genetic map of the linkage group termed cucumber chromosome 2, respectively.

5. The method of claim 1, wherein the allele which confers resistance to Downy Mildew is derived from cucumber line PI197088, or a progeny plant thereof.

6. The method of claim 1, wherein selecting the first cucumber plant further comprises selecting the plant based on the presence of a plurality of genetic markers that map to a position between the sequence represented by SNP marker NN0228457 and SNP marker NN0228148, which map to approximately 25.2 cM and 82.7 cM on the genetic map of the linkage group termed cucumber chromosome 5, respectively;

or wherein the QTL maps to a position between the sequence represented by SNP marker NN0227981 and SNP marker NN0226631, which map to approximately 35.7 cM and 55.5 cM on the genetic map of the linkage group termed cucumber chromosome 5, respectively;

or wherein the QTL maps to a position between the sequence represented by SNP marker NN0227981 and SNP marker NN0247786, which map to approximately 35.7 cM and 71.4 cM on the genetic map of the linkage group termed cucumber chromosome 5, respectively;

or wherein the QTL maps to a position between the sequence represented by SNP marker NN0225012 and SNP marker NN0227587 which map to approximately 20.6 cM and 87.4 cM on the genetic map of the linkage group termed cucumber chromosome 4, respectively;

or wherein the QTL maps to a position between the sequence represented by SNP marker NN0247342 and SNP marker NN0224495 which map to approximately 54.5 cM and 82.0 cM on the genetic map of the linkage group termed cucumber chromosome 4, respectively;

or wherein the QTL maps to a position between the sequence represented by SNP marker NN0223782 and SNP marker NN0226638 which map to approximately 22.5 cM and 88.4 cM on the genetic map of the linkage group termed cucumber chromosome 2, respectively.

7. The method of claim 1, wherein the genetic marker is selected from the group consisting of markers NN0223782, NN0225385, NN0226670, NN0224124, NN0246472, NN0225358, NN0227700, NN0224617, NN0247695, NN0227242, NN0223824, NN0223181, NN0226638, NN0225012, NN0228579, NN0226451, NN0225088, NN0226219, NN0247551, NN0246357, NN0225551, NN0226732, NN0247689, NN0247342, NN0224702, NN0225482, NN0224538, NN0247543, NN0224041, NN0228853, NN0227762, NN0227587, NN0228457, NN0246356, NN0246332, NN0223399, NN0223689, NN0247731, NN0226166, NN0225480, NN0246411, NN0227759, NN0247348, NN0228465, NN0247786, NN0226645, NN0223809, NN0227071, NN0226870, NN0228148, NN0246378, NN0224041, NN0227587, NN5096749, NN0224856, NN0223160, NN0223809, NN0226631, NN0227981, NN0247786, NN0246425, NN0224495, NN0225088, NN0227071, NN0223782, NN0228579, NN0247342, and NN0247731, comprising a single nucleotide polymorphism of one of SEQ ID NOs:

20-87.

8. The method of claim 7, wherein the genetic marker is selected from the group consisting of markers NN0223782, NN0225385, NN0226670, NN0224124, NN0246472, NN0225358, NN0227700, NN0224617, NN0247695, NN0227242, NN0223824, NN0223181, NN0226638, and NN0246378.

9. The method of claim 7, wherein the genetic marker is selected from the group consisting of markers NN0225012, NN0228579, NN0226451, NN0225088, NN0226219, NN0247551, NN0246357, NN0225551, NN0226732, NN0247689, NN0247342, NN0224702, NN0225482, NN0224538, NN0247543, NN0224041, NN0228853, NN0227762, NN0227587, NN0224041, NN0227587, NN0246425, NN0224495, NN0225088, NN0228579, and NN0247342.

10. The method of claim 7, wherein the genetic marker is selected from the group consisting of markers NN0228457, NN0246356, NN0246332, NN0223399, NN0223689, NN0247731, NN0226166, NN0225480, NN0246411, NN0227759, NN0247348, NN0228465, NN0247786, NN0226645, NN0223809, NN0227071, NN0226870, NN0228148, NN5096749, NN0224856, NN0223160, NN0223809, NN0226631, NN0227981, NN0247786, NN0227071, and NN0247731.

11. The method of claim 1, wherein assaying the cucumber plants comprises PCR, single strand conformational polymorphism analysis, denaturing gradient gel electrophoresis, cleavage fragment length polymorphism analysis, TAQMAN assay, and/or DNA sequencing.

12. The method of claim 1, wherein the genetic marker maps within 20 cM, 10 cM or 1 cM of a QTL which confers resistance to Downy Mildew.

13. The method of claim 1, wherein the genetic marker is NN0226631 or NN0246425.

14. A method of cucumber plant breeding comprising:

a) assaying cucumber plants for the presence of at least a first genetic marker genetically linked to a QTL that confers resistance to Downy Mildew; and

b) selecting at least a first cucumber plant comprising the genetic marker and the QTL that confers resistance to Downy Mildew;

wherein the QTL maps to a position between the sequence represented by SNP marker NN0228457 and SNP marker NN0228148, which map to approximately 25.2 cM and 82.7 cM on the genetic map of the linkage group termed cucumber chromosome 5, respectively;

wherein the QTL maps to a position between the sequence represented by SNP marker NN0227981 and SNP marker NN0226631, which map to approximately 35.7 cM and 55.5 cM on the genetic map of the linkage group termed cucumber chromosome 5, respectively;

wherein the QTL maps to a position between the sequence represented by SNP marker NN0227981 and SNP marker NN0247786, which map to approximately 35.7 cM and 71.4 cM on the genetic map of the linkage group termed cucumber chromosome 5, respectively;

wherein the QTL maps to a position between the sequence represented by SNP marker NN0225012 and SNP marker NN0227587 which map to approximately 20.6 cM and 87.4 cM on the genetic map of the linkage group termed cucumber chromosome 4, respectively;

wherein the QTL maps to a position between the sequence represented by SNP marker NN0247342 and SNP marker NN0224495 which map to approximately 54.5 cM and 82.0 cM on the genetic map of the linkage group termed cucumber chromosome 4, respectively;

or wherein the QTL maps to a position between the sequence represented by SNP marker NN0223782 and SNP marker NN0226638 which map to approximately 22.5 cM and 88.4 cM on the genetic map of the linkage group termed cucumber chromosome 2, respectively; and

c) crossing the first cucumber plant with itself or a second cucumber plant to produce progeny cucumber plants comprising the QTL that confers resistance to Downy Mildew.

15. The method of claim 14, wherein selecting at least a first cucumber plant further comprises selecting the plant based on the presence of a plurality of genetic markers that map to a position between the sequence represented by SNP marker NN0228457 and SNP marker NN0228148, which map to approximately 25.2 cM and 82.7 cM on the genetic map of the linkage group termed cucumber chromosome 5, respectively;

wherein the QTL maps to a position between the sequence represented by SNP marker NN0227981 and SNP marker NN0226631, which map to approximately 35.7 cM and 55.5 cM on the genetic map of the linkage group termed cucumber chromosome 5, respectively;

wherein the QTL maps to a position between the sequence represented by SNP marker NN0227981 and SNP marker NN0247786, which map to approximately 35.7 cM and 71.4 cM on the genetic map of the linkage group termed cucumber chromosome 5, respectively;

wherein the QTL maps to a position between the sequence represented by SNP marker NN0225012 and SNP marker NN0227587 which map to approximately 20.6 cM and 87.4 cM on the genetic map of the linkage group termed cucumber chromosome 4, respectively;

wherein the QTL maps to a position between the sequence represented by SNP marker NN0247342 and SNP marker NN0224495 which map to approximately 54.5 cM and 82.0 cM on the genetic map of the linkage group termed cucumber chromosome 4, respectively;

or wherein the QTL maps to a position between the sequence represented by SNP marker NN0223782 and SNP marker NN 0226638 which map to approximately 22.5 cM and 88.4 cM on the genetic map of the linkage group termed cucumber chromosome 2, respectively.

16. The method of claim 14, further comprising the step of

d) selecting a progeny plant comprising the allele which confers resistance to Downy Mildew and crossing the progeny plant with itself or a third cucumber plant to produce additional progeny plants.

17. The method of claim 16, wherein the method further comprises repeating step (d) about 2-10 times.

18. The method of claim 14, wherein the allele which confers resistance to Downy Mildew is derived from cucumber line PI197088, or a progeny plant thereof.

19. The method of claim 14, wherein the genetic marker is selected from the group consisting of markers NN0223782, NN0225385, NN0226670, NN0224124, NN0246472, NN0225358, NN0227700, NN0224617, NN0247695, NN0227242, NN0223824, NN0223181, NN0226638, NN0225012, NN0228579, NN0226451, NN0225088, NN0226219, NN0247551, NN0246357, NN0225551, NN0226732, NN0247689, NN0247342, NN0224702, NN0225482, NN0224538, NN0247543, NN0224041, NN0228853, NN0227762, NN0227587, NN0228457, NN0246356, NN0246332, NN0223399, NN0223689, NN0247731, NN0226166, NN0225480, NN0246411, NN0227759, NN0247348, NN0228465, NN0247786, NN0226645, NN0223809, NN0227071, NN0226870, NN0228148, NN0246378, NN0224041, NN0227587, NN5096749, NN0224856, NN0223160, NN0223809, NN0226631, NN0227981, NN0247786, NN0246425, NN0224495, NN0225088, NN0227071, NN0223782, NN0228579, NN0247342, and NN0247731, comprising a single nucleotide polymorphism of one of SEQ ID NOs:

20-87.

20. The method of claim 19, wherein the genetic marker is selected from the group consisting of markers NN0223782, NN0225385, NN0226670, NN0224124, NN0246472, NN0225358, NN0227700, NN0224617, NN0247695, NN0227242, NN0223824, NN0223181, NN0226638, and NN0246378.

21. The method of claim 19, wherein the genetic marker is selected from the group consisting of markers NN0225012, NN0228579, NN0226451, NN0225088, NN0226219, NN0247551, NN0246357, NN0225551, NN0226732, NN0247689, NN0247342, NN0224702, NN0225482, NN0224538, NN0247543, NN0224041, NN0228853, NN0227762, NN0227587, NN0224041, NN0227587, NN0246425, NN0224495, NN0225088, NN0228579, and NN0247342.

22. The method of claim 19, wherein the genetic marker is selected from the group consisting of markers NN0228457, NN0246356, NN0246332, NN0223399, NN0223689, NN0247731, NN0226166, NN0225480, NN0246411, NN0227759, NN0247348, NN0228465, NN0247786, NN0226645, NN0223809, NN0227071, NN0226870, NN0228148, NN5096749, NN0224856, NN0223160, NN0223809, NN0226631, NN0227981, NN0247786, NN0227071, and NN0247731.

23. The method of claim 14, wherein assaying the cucumber plants comprises PCR, single strand conformational polymorphism analysis, denaturing gradient gel electrophoresis, cleavage fragment length polymorphism analysis, TAQMAN assay, and/or DNA sequencing.

24. The method of claim 14, wherein the genetic marker maps within 20 cM, 10 cM or 1 cM of a QTL which confers resistance to Downy Mildew.

25. The method of claim 14, wherein the genetic marker is NN0226631 or NN0246425.

26. The method of claim 14, wherein the cucumber plant comprising at least one allele which confers resistance to Downy Mildew demonstrates a reduction of foliar symptoms of chlorotic and/or necrotic lesions of at least, or greater than, 25%, relative to a non-resistant control cucumber line.