ISSR Analysis On The Diversity Of Spatholobus Suberectus Germplasms

MillettiareticuiataBenth is mainly refers to the stem of Spatholobus suberectus,but due to people’s unreasonable utilization of resources,wild MillettiareticuiataBent-h resources were being destroyed,and more and more falsify MillettiareticuiataBenth flow into the market,it caused great impactions for the rational use of Millettiareticui-ataBenth.According to the survey of "China pharmacopoeia" in 2005,the medicine source plants of MillettiareticuiataBenth was legume,lardizabalaceae,fructus schisandrae,etc.a total of 6 genera and 15 species and varieties.To further learn MillettiareticuiataBenth,this paper took 12 wild populations with 120 samples about the MillettiareticuiataBenth as material,to carry on the analysis of genetic diversity.The main results were as follows;(1)Based on the actual investigation and sample collection.We record the related morphological features,such as the leaf length,leaf width and so on,as the characteristic properties,and use statistical software to analysis.The results showed that the coefficient of variation for the differences populations was different,so there was genetic variation between populations of species.The analysis of 12 species showed that the relationship of different areas of the same species was close,and the relationship of the same region of the populations was also close.Although phenotypic markers could analysis the genetic relationship and diversity of species,genetic traits determine species phenotype,so to ensure the integrity of the experiment carried out ISSR experiments.(2)According to related literature,established the initial reaction of ISSR-PCR amplification system.The optimal reacting system with the volume of 15μL was established as follows:80ng DNA template,0.8 mmol/L dNTPs,1.2U Taq DNA polymerase,2.0mmol/L Mg2+,0.6μmol/L primers.According to optimization of experimental system,and collected experimental data for analysis of genetic diversity.The result showed that total 221 ISSR discernible bands with 12 ISSR primers.The percentage of polymorphic bands was 94.81%.At the population level with the Shannon’s information index(I),the values were different The highest genetic diversity of populations was Mucuna sempervirens of Hu-nan province and the lowest genetic diversity of populations was Millettia dielsiana Harms of Si-chuan.(3)According to the genetic identity and Nei’s genetic distance,12 species of MillettiareticuiataBenth were analyzed by UPGMA cluster analysis,clustering results showed that the genetic relationship of MillettiareticuiataBenth was more complex.The Spatholobus suberectus populations of Guangxi、Yunnan、Guangdong were being in the same group;the populations of Millettia Metec、Millettia dielsiana Harms、Millettia pachycarpa Benth were being in the same group;the other populations were the other group.The result indicates the relationship between Buji and Wutong Mountain was the closest.The result consistent with phenotypic marker in general.(4)Through the comparative analysis of morphological markers and ISSR molecular markers,found that phenotypic markers was not able to analysis the similar populations of different species,but this method was convenient to analyze the diversity of population.And ISSR molecular markers can avoid plant phenotypic characteristics to analyze the diversity of species.Although it could analysis the population,the repeatability is not high.Two kinds of methods could analyze the diversity of population,but for the related analysis results of consistency was low.