琼脂糖与聚丙烯酰胺凝胶电泳在SSR鉴定杂交油菜种子纯度中的比较

对3对候选引物SR85、SR332、SR407所扩增的秦优33杂交种及其父母亲本的DNA-SSR片段进行琼脂糖与聚丙烯酰胺凝胶电泳分离.两种电泳检测结果比较表明,(1)利用琼脂糖凝胶电泳分离后,3对候选引物对杂交种所扩增的SSR条带均为父母亲本条带的互补.其中引物SR332所获的两条互补条带的迁移率相差大,电泳图谱能清晰鉴别出父本、母本及杂种植株.引物SR85和SR407杂交种的互补条带迁移率相近,在鉴定中易出现人为误差.(2)利用聚丙烯酰胺凝胶电泳分离后,引物SR332所扩增条带未能区别杂交种与父本植株.引物SR85和SR407对杂交种所扩增的条带均为父母亲本所扩增条带的互补,相互差异显著,条带清晰稳定.可见,琼脂糖与聚丙烯酰胺凝胶电泳都是SSR技术鉴定种子纯度的有效途径,种子纯度分析应根据区别材料的遗传关系和引物分析表现来选择不同的电泳介质.

Comparison of Agarose and Polyacrylamide Gel Electrophoresis for SSR in Purity Identification of Qinyou 33 Hybrid Rape Seed

This study have separated the fragments of Qinyou 33 hybrid and parent plants amplified by the three candidate primers SR85,SR332 and SR407 using agarose gel electrophoresis and polyacrylamide gel electrophoresis respectively.The comparative study showed that:(i) Using agarose gel electrophoresis,the bands of hybrid are complemented by the bands of parents amplified by the three candidate primers.The mobility of two bands amplified by the primer SR332 is greatly different.The electrophoresis pattern of SR332 can clearly identify the hybrid and parent plants of Qinyou 33.The complement bands of SR85 and SR407 may mislead the result for their similar mobility.(ii) Using polyacrylamide gel electrophoresis,the bands amplified by the primer SR332 can’t identify the hybrid and male parent plant,but the bands amplified by primer SR85 and SR407 from the hybrid plant are the clear,stable and complement with the different parent plants.This result indicates that agarose gel electrophoresis and polyacrylamide gel electrophoresis is the effective approach to identify the seed purity using SSR.The seed purity analysis should choose different electrophoresis medium basing on the genetic relationship and the primer analysis performance between the difference varieties.