Purity identification of chemical hybriding male varieties of hybrid rape qinyou 33 using ssr markers | 甘蓝型化学杀雄杂交油菜品种秦优33种子纯度的SSR鉴定

Chinese. 为了准确、稳定、高效的鉴定杂交油菜纯度，本研究在简单重复序列(SSR)琼脂糖电泳高效标准分析技术体系基础上，通过对479对引物筛选，获得扩增效果好、条带清晰稳定、可将化学杀雄杂交油菜品种秦优33及其亲本系三者皆可有效区别的SSR引物3对。利用其中的SA332引物对三者的高纯单株分别进行了单株验证，结果表明，三者的SSR单株图谱表现与各自的标准图谱的一致性对应率极高，分别达到了98%、99.33%和96.67%。同时利用SA332和SA85两对引物对秦优33生产商品种种子进行了实用化检测，并和大田形态结果进行了一一定株吻合率比对，结果显示，两对引物与大田的单株吻合率皆能达到96%以上，最高可达98.67%；两引物之间在两样品上的一致性偏差比较仅为0.67%和2.67%；150株室内较小检验群体和大田300株以上较大考察群体的纯度偏差仅为－2.19%到3.76%。研究结果表明，室内SSR的鉴定结果和大田种子纯度的实际表现基本一致，可用于油菜杂种纯度的鉴定。

English. To identify the purity of hybrid rape exactly, steadily and effectively, this study has screened 3 of 479 pairs of SSR primers which could distinguish a chemical hybriding male varieties of hybrid rape Qinyou 33 from its parental lines effectively and steadily based on the establishment of efficient simple sequence repeat (SSR) agarose electrophoresis standard analysis system through the primer screening. The single-plant in three high pure lines was verified by using one of the primer SA332. Results demonstrated that the three single-plant SSR map had high consistency with its corresponding standard maps, reaching 98%, 99.33%and 96.67%, respectively. The practical detection and the consistency of qinyou33 commercial seed compared with single-plant growing in field using the primer SA332 and SA85 were carried out at the same time. The result showed that the consistency compared with single-plant grow in field reached above 96%, 98.67% at most and the consistency between the primer SA332 and SA85 reached 0.67% and 2.67% respectively. The purity deviation between 150 strains of indoor small groups and the field test more than 300 strains only reached -2.19% to 3.76%. With the basically consistency of indoor SSR tests result and field actual purity, it is fully demonstrated that the high consistency SSR correspondence analysis is a powerful tool for identify hybridism purity of rape especially for chemical hybiding male varieties of hybrid rape which are no significant difference in morphology.