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群体遗传学研究对于了解病原菌的流行、变异及进化规律具有重要意义。但是到目前为止,对小麦纹枯病菌禾谷丝核菌群体遗传结构的了解并不深入,缺乏有效的SSR标记是主要原因。本研究根据禾谷丝核菌全基因组序列进行了微卫星位点搜索并设计SSR引物,通过电泳筛选和测序验证,最终筛选出12个多态性较好且稳定可靠的SSR标记。利用这些标记对收集自我国安徽、江苏、河南三省的23个禾谷丝核菌菌株进行了多态性分析,结果发现禾谷丝核菌基因组中长片段微卫星位点较少。12对引物扩增出的等位基因数平均为6.1个,期望杂合度平均为0.651,观测杂合度平均为0.508,表明这些标记具有较高的多态性,能够满足禾谷丝核菌群体遗传学研究需要。本研究为进一步进行禾谷丝核菌的多样性、群体遗传结构分析及进化学研究提供了有效工具。
Population genetic research is important for understanding the prevalence, variation and evolution of pathogens. However, so far, the understanding of the genetic structure of Rhizoctonia cerealis population in R. solani is not deep, and the lack of effective SSR markers is the main reason. In this study, microsatellite loci were searched based on the whole genome sequence of R. cerealis and SSR primers were designed. Through electrophoresis screening and sequencing validation, 12 SSR markers with good polymorphism and stable polymorphism were screened out. Using these markers, 23 strains of Rhizoctonia cerealis collected from Anhui, Jiangsu and Henan provinces in China were analyzed for their polymorphism. The results showed that there were fewer microsatellite loci in the long fragment of R. reinhardtii genome. The average number of alleles amplified by 12 pairs of primers was 6.1, the average expected heterozygosity was 0.651, and the average observed heterozygosity was 0.508, which indicated that these markers had high polymorphism and could meet the population of Rhizoctonia cerealis Study needs. This study provided an effective tool for the further study of Rhizoctonia cerealis diversity, population genetic structure analysis and evolutionary studies.