Genetic Diversity Analysis Of Sophora Alopecuroides And The Correlation Analysis With Matrine

Based on23Sophora alopecuroides L. germplasm resources from Ningxia, Xinjiang,Qinghai, Gansu, and Inner Mongolian in China, genetic diversity, genetic relationship and genetic differentitiation were detected in our research by characterising matrine content, oxymatrine content, SSR and self-designed molecular marker analysis in order to study their genetic background and provide some theoretical evidence for scientifically utilization of S. alopecuroides L. resources, including its protection and collection. The results were as follows:(1) The analysis of matrine content and oxymatrine content in seed and leaf of twenty-three S. alopecuroides L. populations showed that both of the content were differ.The coefficient of variation (CV) of matrine content and oxymatrine content exceeded30%. Matrine content in leaf were2.12times that of the seed, and oxymatrine content in seed were5.36times that of the leaf. Nitrogen fertilization showed a improve of matrine content, and very significant in oxymatrine content.(2) The reaction system of SSR was established and optimized for analyzing the genetic diversity of S. alopecuroides L. populations. The optimized SSR reaction system was as follows:DNA20ng, Taq enzyme0.5U,10×Buffer1μL, Mixed Primer1μmol·L-1, Mg2+2.5mmol·L-1,dNTP0.2mmol·L-1with the supplement of ddH2O to10μl.(3)18SSR primers were selected from30primers according to their reproducible amplification and polymorphisms. The results revealed that the amplification loci was409, polymorphism loci was397. The Nei’s gene diversity and Shannon’s information index were0.2709and0.4150respectively. The genetic distances ranged from0.0883to0.2702. The results of SSR unweighted pair-group method with arithmetic average (UPGMA) analysis demonstrated that23S. alopecuroides L. populations were clustered into6groups.Genetic differentiation coefficiednt was0.4168which means significantly genetic differentiations exist among populations.(4) Designed from sequences of related species checked by NCBI,18self-designed primers were selected from21primers according to their reproducible amplification and polymorphisms. The results revealed that the amplification loci was257, polymorphism loci was214. The Nei’s gene diversity and Shannon’s information index were0.3096and0.4581respectively. The genetic distances ranged from0.2261to0.5451. The results of SSR unweighted pair-group method with arithmetic average (UPGMA) analysis demonstrated that23S. alopecuroides L. populations were clustered into4groups.(5) It’s showed there was no significant correlation between genetic and geographic distances among population in the Mantel test.(6)38loci from36primers associated with the four traits,19and13loci were associated with seed matrine content and leaf oxymatrine content, respectively. Interpretation ratio was0.51and0.48in36D3and13D15, respectively.