The Shiraia Its Separation Of The Genetic Diversity Of Fungal Analysis

Shiraia bambusicola P. Henn. is a parasitic medicinal fungus, which has a very high medicinal value. With the discovery of the photosensitive hypocrellin (hypocrellin A and hypocrellin B), S. bambusicola has increasing got recognition in research and application.In order to understand the genetic differentiation of the medicinal fungus Shiraia bambusicola, the genetic diversity of 107 individuals in 8 populations collected from Jiangsu, Anhui and Zhejiang provinces were studied by using inter-simple sequence repeat(ISSR) analysis. In this study, the effect of different concentrations of Mg2+, dNTPs, Taq DNA polymerase, primer and template DNA was analyzed and the optimized ISSR-PCR reaction system of S. bambusicola was established by the test of the single factoe and orthogonal experiments. Based on the optimized ISSR-PCR reaction system, th genetic diversity and genetic structure of 107 individuals from 8 geographic locations were submitted to analysis. The 11 primers employed produced a total of 241 bands, of which 240 bands (PPB=99.59%) were polymorphic. At population level, the percentage of polymorphic bands waried from 45.23% for Yixing of Jiangsu province to 74.27% for Tianmu Mountain of Zhejiang province, with a mean value of 64.21%. Nei's gene diversity index is 0.3314 and the index of Shannon's genetic diversity was 0.4996, which indicates high level of genetic variability. Genetic differentiation mainly occurred within population (Gst=0.3314). AMOVA detected that variation among populations was 39.96%, variation within populations was 60.04%. Nei's genetic distance between populations ranged from 0.1115 to 0.2453, with a mean value of 0.1766. Gene flow between 8 populations was 1.0086, indicating a little genetic intercourse. The results of unweighted pair group method arithmetic average (UPGMA) analysis and Principal coordinates analysis (PCA) had reveled that the populations with near distance in geography got together frequently and showed regional characteristic. It showed that the relations between sibship and geographical distribution were intensive.In order to identify the related fungi associated with fruiting body of S. bambusicola collected from Yixing in Jiangsu province, Guangde in Anhui province, Anji and Lishui in Zhejiang province,36 fugual strains were isolated from fruiting badies of S. bambusicola. Two molecular phylogenetic trees were generated by neighbor-joining method and maximum parsimony method with the internal transcribed spacers (ITS) of rDNA sequences of the 36 strains and 15 relevant sequences from GenBank. Phylogenetic analysis showed that 29 strains belonged to Ascomycota, 3 strains belonged to Basidiomycota, and 4 stains belonged to Deuteromycotina. In Ascomycota, the members of the Pleosporales and Apiosporaceae were dominant groups, secongdly including the Hypocreales and Sordariales.