水稻种质资源指纹与多样性分析的分子标记体系研究

Molecular Marker Systems for Fingerprinting and Diversity Analysis of Rice Germplasm Resources

应用2个抗性基因同源序列(RGA)标记:C-末端富亮氨酸重复序列(CLRR)与核糖核酸酶抑制因子亮氨酸重复序列(RLRR),及2个Seoulin研究所生命科学通用引物标记(SRILS):SRILS6与SRILS8,通过这4个分子标记所产生的指纹图谱,菲律宾国家水稻所水稻种质资源库中265个地方品种的遗传差异及亲缘关系得到了确立。DNA指纹图谱结果显示,引物CLRR分辨能力最高,共扩增出了48条多态带,平均每个品种28.6条,因而是快速DNA指纹图谱分析最有效的标记。从群体整体水平上来看,RGA与SRILS两套引物对265份材料的平均多态带率分别为60.6%与60.5%,表明两种标记体系具有相似的多态性检测能力。根据4个引物的DNA指纹谱带数据,进行单个及合并聚类分析,计算出7个极为相似的聚类系统树图:将样品主要聚成两个大类群,类群Ⅰ与Ⅱ。聚类分析的结果表明,RGA与SRILS标记体系各自均能完全鉴别出所有265个品种,而且揭示了一个相对高的群体遗传多样性。7个聚类系统树图的聚类分布以及类群的组成均极为相似,有88%～97%的相似度,这说明采用的RGA及SRILS引物扩增的基因组区域可能已经发生了一定程度的变异以致表现出高度一致的遗传系谱。

The genetic variability and relationships among 265 rice germplasm accessions from the Genebank of Philippine Rice Research Institute were established from DNA fingerprints generated by two resistance gene analogue（RGA） markers： C-terminal leucine-rich repeat（CLRR） and ribonuclease inhibitor leucine-rich repeat（RLRR）,and two universal primer markers： Seoulin Research Institute Life Science-6（SRILS6） and Seoulin Research Institute Life Science-8（SRILS8）.The DNA fingerprints showed that CLRR was the most discriminating marker,generating 48 polymorphic bands in the germplasm samples with an average of 28.6 bands per accession,thus it was the most efficient marker for rapid DNA fingerprinting analysis.On the average,the RGA and SRILS primer markers showed the similar power in detecting the polymorphism within the population at the entire germplasm population level,and the average percentage of polymorphic bands generated by RGA and SRILS primer markers was 60.6% and 60.5%,respectively.Seven dendrograms were generated based on the banding profiles generated by RGA and SRILS primers or their combinations,featuring two major groups： group Ⅰ and group Ⅱ.Dendrogram analysis showed that both RGA and SRILS primers could distinguish all 265 accessions,and revealed a high level of diversity among the accessions.The clustering patterns and the compositions of the groups had the similarity of 88%~97% among the seven dendrograms.This indicated that the genomic regions amplified by these particular RGA and SRILS markers might have undergone mutations at comparative rate so as to manifest highly consistent genetic trees.