基于差减cDNA文库EST信息的月季花香突变体SSR标记的开发

Development of new SSR markers from EST of SSH cDNA libraries on rose fragrance

在前期月季花香突变体差减文库EST序列信息的工作基础上,文章开发出新的与花香相关的SSR标记。从正反向差减文库391条EST中检索到10条含有10个SSR的序列,SSR的检出率为2.6%,EST-SSR的重复基元共搜索到10种。利用部分EST-SSRs序列设计了6对SSR引物,以花香突变体‘往日情怀’及其野生型‘金银岛’DNA为模板,对引物进行筛选,5对引物有扩增条带,其中3对引物有特异性扩增条带。同时利用这些可扩增的引物对典型芳香和无香两组月季栽培品种进行多态性检测,发现这5对引物均显示多态性。表明所建立的SSR标记是一种可行而有效的方法。

The new SSR markers of rose related fragrance were developed based on the SSH cDNA libraries of rose floral scent mutant. In this study, 10 EST-SSRs （2.6%） from 391 ESTs in the libraries were identified. Six EST-SSRs primers were designed to sequence flanking SSRs. The primer pairs designed were screened on the wild-type Jinyindao, which has flowers full of pleasant scent, and the mutant-type Wangriqinghuai without perceivable floral scent. Five primer pairs were amplified effectively in Jinyindao and Wangriqinghuai, and 3 were polymorphic between Jinyindao and Wangriqinghuai. Eighteen rose cultivars including fragrant roses and nonfragrant roses were identified by the five prime pairs. These results proved that EST-SSR markers are effective markers to identify the polymorphism of the rose.