甜瓜分子标记纯度鉴定研究

Identification of molecular marker of purity in melon

以2个甜瓜品种西域雪1号、西域雪2号及其亲本为试材,建立并优化了单粒种子DNA快速提取法和适合甜瓜的RAPD、SSR分子标记杂交种纯度鉴定体系。从100条随机引物中筛选出G17用于西域雪2号纯度鉴定,可有效区分母本和杂交种。从19对SSR引物中筛选出9对稳定扩增、多态性丰富的引物构建西域雪1号、西域雪2号的SSR指纹图谱。SSR分子标记用较少引物即可扩增出品种特异性条带,且多数F1代带型呈双亲互补,非常适合种子纯度鉴定。同时,使用高浓度、非变性的聚丙烯酰胺凝胶电泳、银染,条带更清晰,可区分只相差1个碱基的扩增片段,同RAPD相比,SSR分子标记更适合甜瓜杂交种纯度快速鉴定。

Two melon hybrids Xiyuxue No. 1,Xiyuxue No. 2 and their parents were used to establish the system of rapid DNA extraction method, the suitable melon RAPD, SSR and the identification of molecular marker of purity in melon. The random primer, G17, out of 100 pair of primers was screened to identify the purity of Xiyuxue No. 2. The band type resembled paternal line. Nine pairs of SSR primers with steady and polymorphism band were screened from nineteen pairs of primers to establish fingerprint of Xiyuxue No. 1 and Xiyuxue No, 2, It can amplify specific band with different hybrids by using less SSR primers, Numerous band type presents parent complement. Meanwhile, after nondenaturing high-desity PAGE gel electrophoresis and silver nitrate stains, the bands were clearer, only one bp difference can be found. Compared with RAPD, SSR molecular marker is more suitable to identify purity of melon hybrids rapidly.