通过提取细胞质雄性不育系和同型保持系的线粒体基因组DNA,用100条ISSR引物进行筛选,发现引物ISSR829的扩增片段在不育系和保持系间存在差异。对差异片段进行测序发现,该序列仅存在于保持系线粒体基因组中,且位于基因Atp6上游2 000 bp处,推断其可能与不育有关。利用测序获得片段设计引物,用目前已育成的5个大豆杂交种的不育系母本和同型保持系的基因组DNA为模版,进行PCR扩增,发现不育系母本未出现特异片段,而对应保持系出现目的片段,表明该特异片段转化为SCAR标记,可用于不育系的筛选和不育系种子纯度鉴定。 Mitochondrial genomic DNA was extracted from cytoplasmic male sterile line and homogenous maintainer line by 100 ISSR primers. The amplified fragment of primer ISSR829 was found to be different between male sterile line and maintainer line. Sequencing of the difference fragments found that the sequence was only present in the maintainer mitochondrial genome and was located at 2 000 bp upstream of Atp6, suggesting that it may be related to infertility. The fragment was designed by sequencing, and the genomic DNAs of the male sterile line and the maintainer line of the five soybean hybrids that have been bred at present were used as templates for PCR amplification. No specific fragments were found in male sterile lines of male sterile line Corresponding maintainer lines showed the objective fragment, indicating that the specific fragment was transformed into a SCAR marker, which can be used for screening of sterile lines and identification of seed purity of sterile lines.