杂交油菜宁杂11号种子纯度SSR标记快速检测方法

Rapid Purity Identification of Hybrid Rapeseed Ningza No.11 by Using SSR Markers

本研究以杂交油菜新品种宁杂11号为研究对象,利用SSR分子标记技术,筛选出共显性标记引物Na10-E02,建立了一套油菜杂交种纯度的快速分子检测方法:种子利用夜间萌发,碱裂解法快速提取DNA,PCR反应2h,电泳1.5h,简化的银染法染色10min,从获取样品种子到出检测结果只需要不到一天的时间,一个熟练科技人员每个工作日至少可以完成6×96=576粒种子的检测量。利用这套技术对生产中大面积制种的9个样品纯度检测结果与田间种植鉴定的实际纯度结果极显著正相关,相关系数达到0.984(P<0.01),表明这套技术可以用于宁杂11号杂种纯度的快速准确鉴定。

In this study, the new rapeseed hybrid combination Ningza No.11 as experimental materials, a set of rapid molecular identification techniques for rapeseed hybrid purity were established by using SSR molecular markers to screen co-dominant marker primer Na10-E02, The processes were including： seed germination in night, DNA extraction by alkaline lysis method, the PCR amplification for 2 hours, gel electrophoresis in 1.5 hours, and simplified silver staining in 10 minutes. The test result for hybrid purity identification came out within a day from seed sampling, so a skilled technician could at least complete 576 seed samples a day （6×96=576）. By using this process and combining the field validation, nine sample stocks that large scale applied in production were employed to be identified their purity, The results exhibited highly significant positive correlation between two methods, the correlation coefficient reached 0.984 （P0.01）, indicating that this technology can be applied to rapid and accurate identify hybrid purity of the Ningza No.11.