Development of allele-specific SNP markers for the new low-linolenic mutant of winter oilseed rape

摘要

Genotypes of oilseed rape characterized by lowered linolenic acid content in seed oil are under development and investigation worldwide due to the non-food use of rapeseed oil for biofuel biocomponents production. The low linolenic mutant (LLMut), characterized by about 2% of linolenic acid in seed oil, was obtained as a result of chemical mutagene treatment of double low (00) winter oilseed rape breeding line (about 10% of linolenic acid in seed oil) at the Plant Breeding and Acclimatization Institute in Poznan, Poland. The aim of this work was to develop allele-specific DNA markers for the obtained genotype. Genomic DNA was isolated from the low-linolenic mutant and from non-mutated (00) breeding line. Fad3 desaturase gene alleles of LLMut and 00 genotypes were PCR amplified, cloned and sequenced. Two significant mutation changes in nucleotide sequences were identified and allele-specific SNP markers were designed. DH lines population was developed from F1 hybrid obtained as a result of 00 × LLMut crossing. SNP genotyping of the DH lines was done and correlation coefficient (r) was calculated, indicating a strong negative correlation between linolenic acid content in seed oil and the presence of the mutations in fad3 gene.