粗皮侧耳栽培种EST-SSR分子标记的开发应用及初级核心种质库的构建

根据已知的粗皮侧耳、灵芝、香菇EST序列开发出9条粗皮侧耳EST-SSR引物、58条灵芝EST-SSR引物和35条香菇EST-SSR引物,从中筛选出4条多态性好、条带清晰的引物,对选取的185株粗皮侧耳栽培种进行扩增分析。共扩增得到59条带,每个引物扩增条带数为12–19条,扩增片段在100–1,100bp之间。根据扩增结果构建了一个含47株粗皮侧耳栽培种的初级核心种质库P-core47。分析得出P-core47占原群体样品数25%,占资源库的13%,有效等位基因数和多态性位点占有率与原群体一致,多样性指数均值比原群体高0.12747,在原群体的低频率等位基因有所增强。P-core47内菌株来源广泛,基本已覆盖各个栽培地区,统计的质量性状和数量性状均可在其中找到代表性菌株。表明了用EST-SSR分子标记构建粗皮侧耳栽培种初级核心种质库的可行性。

Development of EST-SSR markers in Pleurotus ostreatus and construction of primary core collection

Based on the EST sequences published on NCBI, 9 Pleurotus ostreatus EST-SSR primer pairs, 58 Ganoderma lucidum EST-SSR primer pairs and 35 Lentinula edodes EST-SSR primer pairs were designed. Among these, 4 of good polymorphisms and clear strips were selected to perform the amplification analysis for the cultivation of the chosen 185 strains of Pleurotus ostreatus. After amplification, 59 alleles were generated, and there were 12–19 alleles for each primer, with the fragment sizes between 100bp and 1,100bp. A primary core collection, P-core 47, composed of 47 accessions was constructed based on the amplification results. After analysis, P-core 47 accounts for 25% in the original sample pool, 13% in the oringinal resource pool; as for the number of effective alleles and occupation ratio of polymorphism sites, consistency was preserved between P-core 47 and the original pool; in addition, compared to the original pool, diversity index of P-core outperformed by 0.12747, and alleles used to be of low frequency in the original pool were strengthened as well. The strains in P-core 47 are widely-ranged, basically available in most of the cultivation regions; you can find representative alleles of both qualitative and quantitative characters in this collection. This showed the feasibility of the adoption of EST-SSR markers to construct a primary core collection of the Pleurotus ostreatus.