杂交油菜宁杂11号种子纯度SSR标记快速检测方法(英文)

目的]建立杂交油菜宁杂11号种子纯度SSR标记快速检测方法。方法]本研究以杂交油菜新品种宁杂11号为研究对象,利用SSR分子标记技术建立杂交种纯度快速检测方法,同时结合田间种植鉴定作为对比验证。结果]筛选出共显性标记引物Na10-E02,建立了杂交油菜宁杂11号种子纯度SSR标记快速检测方法:种子利用夜间萌发,碱裂解法快速提取DNA,PCR反应2 h,电泳1.5 h,简化的银染法染色10 min,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(P0.01)。结论]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。

A Method for Rapid Identification of Ningza 11 Seeds Purity with SSR Markers

Objective] This study aimed to establish a method for rapid identification of Ningza 11 seeds purity with SSR markers. Method] Taking Ningza 11 hybrid seeds as experimental materials, a method for rapid identification of hybrid rape-seeds was established with SSR molecular markers; meanwhile, the test seeds were planted in the field for comparison and verification. Result] A method for rapid identification of Ningza 11 seeds purity with SSR molecular markers was estab-lished： DNA from seeds germinated in the night was extracted by alkaline lysis method; the PCR amplification was performed for 2 h, and electrophoresis for 1.5 h, and a silver staining for 10 minutes. It took less than one day to from obtaining sampling seeds to obtaining the purity identification result, so a skil ed professional can complete the detection of at least 6 ×96 = 576 seeds per weekday. By using this set of detection system, the measured purity of seeds from nine samples was extremely significantly positively correlated to the actual purity identified in the field test, with a correlation coefficient of up to 0.984 （P〈0.01）. Conclusion] This SSR-PCR molecular identification system can be applied for rapid and accurate identifi-cation of Ningza 11 hybrid seeds.