基于油菜F_1杂种种皮鉴别其母本的SSR检测方法及应用

SSR Identification Technology of Female Parent of Brassica napus L.F_1 Hybrids Based on Testa and Its Application

甘蓝型油菜杂交种中雄性不育母本假杂种是影响种子纯度的关键。为了探索一种能准确鉴定油菜杂交种中母本假杂种的方法,利用CTAB小样法分别提取油菜F1杂种种皮和幼苗的总DNA,进而筛选种皮与幼苗的SSR谱带存在差异的引物,并用于鉴别杂交种制种样品中母本基因型假杂种。结果表明:F1杂种种皮的SSR电泳谱带与其母本相同,利用筛选到的多态性SSR引物对2个国审品种的6份杂交制种样品中母本基因型假杂种鉴定的结果与田间单株鉴定吻合率达98%以上,说明利用F1杂种种皮与幼胚存在多态性的SSR标记鉴定杂交种的母本基因型假杂种是可靠的。该技术方法可以克服目前油菜杂交种SSR纯度鉴定对亲本的依赖,为种子生产、管理和经营部门种子纯度质量的有效监控提供技术参考。

Female parent purity is crucial for affecting hybrid seed purity in Brassica napus hybrid seedsproduction. To investigate an effective method suitable to the identification of the false hybrid sample withgenotype of female parents in hybrids samples, this paper adopted CTAB method to extract the total DNA ofhybrid F1 seedlings and its seed coats respectively. Then, the DNA was used to screen the SSR primers whichcould distinguish the hybrid seeds from female parents, and the primers obtained were used to identify the falsehybrid sample with genotype of female parents in hybrids samples. The results showed that hybrid F1 seedlingsand its seed coats had the same SSR fingerprinting, and the coincidence rate was over 98% between the SSRdetection results obtained by the method in this study and field performance, this results indicated that themethod for detecting the false hybrid sample with genotype of female parents in hybrids samples was reliable.The method of this research can overcome the dependence of current SSR identification rapeseed hybridspurity on its parents, provide technical reference on seed production and seed purity quality controlmanagement of business departments.