Soybean is one of the most widely planted crops supplying oil and edible bean. Soybean has a complicated genome due to at least two rounds of polyploidization. Although there are several types of molecular markers available for soybean genotyping, agarose gel-based molecular markers for this purpose are not sufficient. The objective of this study was to develop STS markers that can discriminate Korean soybean cultivars using agarose gels. We employed methylation filtering (MF)technique for constructing a genomic DNA library of “HwangKeum” and selected 1,440 colonies. Of the 1,440 colonies, 887colonies which had more than 700 bp of insert DNA were sequenced. A total of 143 STS primer sets were designed from sequencing data of selected colonies. One hundred one STS primer sets showed unique band in 14 Korean soybean cultivars and digested with 7 restriction enzymes including Hinf I. Among them, 18 STS primer sets showed polymorphic bands. We developed 51 marker combinations and selected 14 marker combinations, which were used to discriminate 110 Korean soybean cultivars.
As the results, 106 cultivars (96.4%) were discriminated by using agarose gel-based markers. Therefore, STS-CAPS markers could be useful to discriminate soybean cultivars and identify soybean genotypes.