利用InDel标记鉴定化学杀雄杂交油菜‘秦油88’种子纯度

Purity Identification of Chemical Emasculation Hybrid Rape(Brassica napus)’Qinyou88’with InDel Markers

In Del(Insertion deletion length polymorphism)是根据近缘物种不同个体间基因组同一位点的序列发生不同大小DNA片段的插入或缺失(Insertion-deletion)而设计的引物进行PCR检测,其以丰富的多态性、操作简单性、结果可靠性而被越来越多地应用于作物遗传研究中。本研究利用InDel标记对‘秦油88’及亲本进行分析,获得了四对带型互补、扩增结果稳定的鉴定‘秦油88’种子纯度的候选引物。利用高纯度杂交种和亲本对其中2对引物ID2和ID15进行验证,一致性分别达到95.5%、100%和100%,97.5%、99.5%和100%。对田间鉴定结果与分子标记鉴定结果进行比较,ID2和ID15引物与田间鉴定结果平均为96.14%和95.98%,同时,两对引物之间的分析偏差仅为0.84,分子鉴定结果与田间鉴定结果及两对引物之间吻合率极高,达到了极显著水平。结果表明,应用InDel分子标记技术进行‘秦油88’种子纯度分析是可行且可靠的。

In Del(Insertion deletion length polymorphism)markers is designed for PCR detection based on the Insertion or deletion of DNA fragments of different sizes according to the sequence of a locus between different individuals of related species,which is more and more applied in crop genetic research due to its rich polymorphism,operational simplicity and reliability of results.In this study,’Qinyou88’and the parents were analyzed through InDel marker,and four pairs of candidate primers were obtained to identify the seed purity of’Qinyou88’with complementary band type and stable amplification results.Two pairs of primers ID2 and ID15 were verified through identified the high purity hybrids and parents,and the consistency reached 95.5%,100%and 100%;97.5%,99.5%and 100%,respectively.By comparing field identification results with molecular marker identification results,the average identification results of ID2 and ID15 primers and field identification results were 96.14%and95.98%,nevertheless,the analysis deviation between the two pairs of primers was only 0.84.The consistency rate between the molecular identification results and field identification results and the two pairs of primers was very high,reaching an extremely significant level.The results show that it is feasible and reliable to analyze the seed purity of’Qinyou88’using InDel molecular marker.