Selection and establishment of high oleic acid content in soybean by using their molecular markers

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The mutants KK21 and HOLL, both of which contain 40 to 45% oleic acid of total fatty acids, were reciprocally crossed in our laboratory. Six high oleic acid lines (approx. 60% of total fatty acid) were selected from F2 progeny and were further confirmed by analyzing the fatty acid contents in both of F3 and F4 progenies.As a result, three of the six selected lines showed higher oleic acid phenotype, compared with their parental lines. The oleic acid content of these three lines distributed in the range of 57.9 to 67.6%, 56.4 to 68.4% and 47.4 to 68.4%, respectively. These values were relatively higher than each of their parents, KK21 (37.2 to 51.2%) or HOLL (44.6 to 53.8%), which were grown under same condition. Furthermore, we selected 3 to 5 F3 offspring-lines from each of 3 lines and those 11 offspring-lines that consist of 20 plants of F4 progeny, were subjected to analyze for oleic acid content. We found one of these lines that has very high oleic acid content which is more than 60%.In addition, we carried out a comparison study for the expression levels of FAD2-1 or FAD2-2 with northern-blot and quantitative RT-PCR techniques, but no significant difference was detected between Bay (original variety) and other mutants. To determine their mutation at the base substitution level, we are now analyzing the nucleotide sequences of FAD2-1 and FAD2-2 gene, that were isolated from Bay and other mutants.It is necessary to push forward the examination of genetic stability of very high oleic acid content of selected one using the next generation, and to analyze the genotype of this selected germline with the designed primers that can detect the polymorphism between Bay and other mutants.