荔枝SSR标记的研究

以无核荔枝A4号为实验材料，应用选择性扩增微卫星（SAM）法分离、克隆了100个简单序列重复（SSR）序列，其中88个非重复，可用。加上搜索数据库所获得的1个SSR序列，一共89个序列用于特异引物的设计。仅从71个序列的82个基因座设计出特异引物。合成41条特异引物(与5′锚定简并引物配对，个别相互配对)，对其中的39个基因座进行检测。其中15对引物扩增出相应大小的片段，另外11对引物扩增出非预期片段。最后，以37个荔枝种质的基因组DNA为模板，从26对出带的引物中，筛选出多态性引物21对，获得了22个荔枝基因座特异性SSR标记。

Development of SSR Markers in Litchi( Litchi chinensis)

A total of 100 SSR sequences were isolated and cloned by means of SAM (Selectively Amplified Microsatellite)techniques and another one was obtained by searching the NCBI and EMBL databases. There were 89 SSR sequences used for design of special primers. As a result, the primers were designed at 82 loci from 71 fragments. Forty-one special primers were synthesized, pairing with 5'anchored degenerate SSR primer, to detect 39 SSR loci. Fifteen of them amplified the corresponding SSR sequences and Other 11 SSR primer pairs amplified non-expected fragments. In the end, 21 polymorphic primer pairs were selected from 26 primer pairs which amplified clear and robust DNA fragment by using the genome DNA of 37 litchi germplasm materials, and 22 locus-specific SSR markers were obtained.