Molecular markers for blackleg resistance gene RLM7 in
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 Rlm7 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 marker assisted selection to produce a progeny Brassica napus plant or germplasm that comprises Rlm7 locus for blackleg resistance, the method comprising:
assaying isolated nucleic acids from a first Brassica napus plant or germplasm for genomic markers thereof;
detecting in the isolated material at least one marker linked to Rlm7 locus for blackleg resistance, wherein the at least one marker is a single nucleotide polymorphism (SNP) selected from the group consisting of DBSNP216588 shown in SEQ ID NO:11, DBSNP216601 shown in SEQ ID NO:12, DBSNP216602 shown in SEQ ID NO:13, DBSNP216603 shown in SEQ ID NO:14, DBSNP216604 shown in SEQ ID NO:15, DBSNP216600 shown in SEQ ID NO:16, DBSNP216606 shown in SEQ ID NO:17, DBSNP216607 shown in SEQ ID NO:18, and DBSNP216608 shown in SEQ ID NO: 19;
selecting the first Brassica napus plant or germplasm thereof based on the detection of the at least one marker linked to blackleg resistance in its isolated nucleic acids; and
crossing the selected first Brassica napus plant or germplasm with a second Brassica plant or germplasm to thereby produce progeny Brassica napus plant or germplasm comprising the at least one marker linked to blackleg resistance.
2. The method according to claim 1 , wherein the detected marker displays a genetic recombination frequency of less than about 10% with the at least one other Rlm7 locus SNP marker of DBSNP216588 shown in SEQ ID NO:11, DBSNP216601 shown in SEQ ID NO:12, DBSNP216602 shown in SEQ ID NO:13, DBSNP216603 shown in SEQ ID NO:14, DBSNP216604 shown in SEQ ID NO:15, DBSNP216600 shown in SEQ ID NO:16, DBSNP216606 shown in SEQ ID NO: 17, DBSNP216607 shown in SEQ ID NO:18, and DBSNP216608 shown in SEQ ID NO: 19, wherein the detected marker and the other marker are different markers.
3. The method according to claim 1 , wherein the detecting comprises amplifying the marker or a portion of the marker to yield an amplified marker amplicon, and detecting the resulting amplified marker 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 isolated material, wherein the primer or primer pair is complementary or partially complementary to at least a portion of the marker, and is capable of initiating DNA polymerization by a DNA polymerase using the isolated 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 4 , wherein the nucleic acid is RNA.
6. The method according to claim 3 , wherein the amplifying comprises utilizing a polymerase chain reaction (PCR) or ligase chain reaction (LCR) using a nucleic acid isolated from the first canola plant or germplasm as a template in the PCR or LCR.
7. A method for producing an introgressed Brassica napus plant or germplasm, the method comprising:
crossing a first parent Brassica napus plant or germplasm thereof with a second parent Brassica napus plant or germplasm thereof to produce a progeny Brassica napus plant or germplasm thereof, wherein the first Brassica napus plant or germplasm comprises at least one single nucleotide polymorphism (SNP) marker allele for blackleg resistance, wherein the at least one SNP marker is selected from the group consisting of DBSNP216588 shown in SEQ ID NO:11, DBSNP216601 shown in SEQ ID NO: 12, DBSNP216602 shown in SEQ ID NO: 13, DBSNP216603 shown in SEQ ID NO:14, DBSNP216604 shown in SEQ ID NO:15, DBSNP216600 shown in SEQ ID NO:16, DBSNP216606 shown in SEQ ID NO: 17, DBSNP216607 shown in SEQ ID NO:18, and DBSNP216608 shown in SEQ ID NO: 19;
isolating nucleic acids from the progeny Brassica napus plant or germplasm thereof;
and detecting the presence of at least one marker for blackleg resistance in the isolated nucleic acids thereby producing an introgressed Brassica napus plant or germplasm having the at least one marker for blackleg resistance.
8. The method according to claim 7 , wherein the second parent Brassica napus plant or germplasm displays lower blackleg resistance as compared to the first parent Brassica napus plant or germplasm, and wherein the progeny plant or germplasm displays an increased resistance to blackleg disease as compared to the second parent plant or germplasm.
9. The method according to claim 7 , wherein the second parent Brassica napus plant or germplasm is a plant or germplasm from an elite canola variety or an exotic canola variety.
10. The method according to claim 1 , wherein the method comprises electronically transmitting or electronically storing data representing the detected marker in a computer readable medium.