Molecular markers for blackleg resistance in canola and methods of using the same
This disclosure concerns methods and compositions for identifying canola plants that have a blackleg resistant phenotype. Some embodiments concern molecular markers to identify, select, and/or construct blackleg resistant plants and germplasm, or to identify and counter-select plants that are susceptible or have low resistance to blackleg disease. Some embodiments concern molecular markers to identify, select, and/or construct blackleg resistant plants that carry the rlm4 gene. This disclosure also concerns canola plants comprising a blackleg resistant phenotype that are generated by methods utilizing at least one marker described herein.
1. A method for identifying a first parent canola plant or germplasm that comprises a marker allele for blackleg resistance in a breeding program, the method comprising:
obtaining a nucleic acid sample from a first parent canola plant or germplasm;
detecting in the nucleic acid sample at least one single nucleotide polymorphism (SNP) marker allele for blackleg resistance selected from the group consisting of DBSNP10503 (SEQ ID NO:24), DBSNP10504 (SEQ ID NO:25), DBSNP31606 (SEQ ID NO:23), DBSNP30220 (SEQ ID NO:22), DBSNP01654 (SEQ ID NO:26), DBSNP01910 (SEQ ID NO:27), DBSNP05704 (SEQ ID NO:28), DBSNP05705 (SEQ ID NO:29), DBSNP28066 (SEQ ID NO:17), DBSNP27644 (SEQ ID NO:18), DBSNP28099 (SEQ ID NO:19), DBSNP33158 (SEQ ID NO:20), BSNP14607 (SEQ ID NO:21), DBSNP04906 (SEQ ID NO:31), DBSNP07219 (SEQ ID NO:30), DBSNP08872 (SEQ ID NO:32), DBSNP08485 (SEQ ID NO:34), DBSNP00547 (SEQ ID NO:33), DBSNP08169 (SEQ ID NO:35), DBSNP00787 (SEQ ID NO:36), and DBSNP01590 (SEQ ID NO:37);
selecting the first parent canola plant or germplasm thereof based on the presence of the SNP marker allele for blackleg resistance in the sample; and
crossing the first parent canola plant with a second parent canola plant to produce a population comprising progeny canola plants or germplasm having the SNP marker allele.
2. The method according to claim 1 , wherein detecting the SNP marker allele comprises use of a probe.
3. The method according to claim 1 , wherein detecting the SNP marker allele comprises amplifying nucleic acid in the sample to produce an amplicon comprising the SNP marker allele, and detecting the SNP marker allele in the amplicon.
4. The method according to claim 3 , wherein the amplifying comprises:
admixing an amplification primer or amplification primer pair with a nucleic acid isolated from the first parent canola plant or germplasm, wherein the primer or primer pair is complementary to at least a portion of genomic DNA comprising the SNP marker allele, and is capable of initiating DNA polymerization by a DNA polymerase using the canola nucleic acid as a template; and
extending the primer or primer pair in a DNA polymerization reaction comprising a DNA polymerase and a template nucleic acid to generate at least one amplicon.
5. The method according to claim 3 , wherein the amplifying comprises utilizing a polymerase chain reaction (PCR) or ligase chain reaction (LCR) using the nucleic acid sample from the first parent canola plant or germplasm as a template in the PCR or LCR.
6. The method according to claim 1 , wherein the second parent canola plant or germplasm is a plant or germplasm from an elite canola variety or an exotic canola variety.
7. The method according to claim 1 , wherein the method further includes harvesting seed from one or more progeny canola plants comprising the SNP marker allele.
8. The method according to claim 1 , further comprising
detecting the SNP marker allele in one or more of the progeny canola plants or germplasm thereof;
selecting one or more progeny canola plants having the SNP marker allele; and
selfing or crossing the one or more selected progeny canola plants.
9. The method according to claim 1 , wherein the method comprises
bulking seed from the next generation of plants produced by the selfing or crossing of the one or more selected progeny canola plants.