基于SRAP分子标记的23株大球盖菇遗传多样性和亲缘关系分析

Genetic Diversity and Phylogenetic Relationship Analysis of 23 Stropharia rugosoannulata Strains Based on SRAP Molecular Marker

利用相关序列扩增多态性(sequence-related amplified polymorphism,SRAP)分子标记对国内外不同来源的23株大球盖菇菌株进行遗传多样性和亲缘关系分析,从288对引物中筛选出稳定性好、可重复性高、多态性稳定并能扩增出清晰明亮条带的38对引物。通过SRAP扩增图谱分析得出,以基因组DNA为模板的23株菌株扩增片段分子量在100~2 000 bp间,共扩增出322个位点,其中有205个多态性位点,多态位点比率为63.66%;供试菌株间的遗传相似系数为0.6646~0.9658;采用UPGMA法对扩增图谱进行聚类分析,在相似水平0.84时可将23株菌株分为3个类群,第Ⅰ类群为来自国外的2株菌株;第Ⅱ类群为本实验室驯化筛选的3株耐高温品种,第Ⅲ类群为国内不同地市的18株菌株。结果表明,不同大球盖菇菌株间存在明显遗传多样性,SRAP分子标记技术可将23株供试菌株鉴别开,且能分析出不同菌株间的亲缘关系。本研究可为大球盖菇菌株亲缘关系的鉴定及新品种选育提供理论依据。

Sequence-related amplified polymorphism( SRAP) molecular marker technique was used to study the genetic diversity and phylogenetic relationship among 23 Stropharia rugosoannulata strains collected from home and abroad. A total of 288 pairs of SRAP primers were tested,and 38 pairs could produce high-quality PCR production with good stability,repeatability and polymorphism. The analysis of SRAP amplification pattern showed that 322 bands were amplified from genomic DNA of 23 strains with the size of 100 ~ 2 000 bp,of which 205 bands were polymorphic and the percentage of polymorphic sites was 63. 66%. The genetic similarity coefficient among the tested strains was ranged from 0. 6646 ~ 0. 9658. The cluster analysis proceeded by UPGMA method showed that 23 strains could be divided into three major groups at the similarity coefficient of 0. 84. Group Ⅰ included two strains from abroad; Group Ⅱ included three high temperature resistant strains domesticated by our laboratory; Group Ⅲ were 18 strains cultivated or preserved in different cities in China. The results showed that significant genetic diversity existed among the strains. SRAP molecular marker technique could identify 23 strains clearly and analyze the phylogenetic relationship among different strains.This test provides the theoretical basis for the identification of Stropharia rugosoannulata phylogenetic relationship and new strain breeding.