Analysis On Genetic Diversity Of Different Populations Of Lespedeza Davurica In Shanxi Province

In this research, RAPD markers, ISSR markers and AP-PCR markers were used to analyze the genetic diversity of23wild Lespedeza davurica in Shanxi Province. Among195random primers,61high polymorphic bands were selected to study the genetic diversity at the molecular level. Defined genetic differences between different populations of Lespedeza davurica initially, provided the basis to develop and take advantage of germplasm resources of Lespedeza davurica.RAPD markers, ISSR markers and AP-PCR markers were widely used in genetic diversity studies for the advantages of simple, convenient, cost-saving, no species-specific and so on. In this research, RAPD, ISSR and AP-PCR markers were combined cluster analysis of23Lespedeza davurica with genetic diversity study. The main results are as follows:1. The establishment of Lespedeza davurica RAPD system:in25μL RAPD amplification reaction system, including DNA template60ng, Mg2+1.75mmol/L(final concentration), dNTP0.3mmol/L, random primer0.4μmol/L, Taq DNA polymerase1U, lOxBuffer2.5μL.14RAPD primers selected from the Shanghai Biological Engineering Company195random primers.217polymorphic fragments obtained for all,196were polymorphic, accounting for90.3%,16fragments for every primer. Based on RAPD analysis of14RAPD primers of Lespedeza davurica, genetic similarity among populations of the GS range is0.56to0.88, with an average of0.73. According to the genetic similarity coefficient system clustering tree diagram, when GS=0.62to0.65,23populations of Lespedeza davurica were divided into two categories:the populations of1to8,10to23were divided into one, population9falled into the another.2.The establishment of Lespedeza davurica ISSR system:volume of ISSR amplification reaction system is25μL, DNA template35ng, Mg2+2.0mmol/L(final concentration), dNTP0.6mmol/L, primer0.4μmol/L, Taq DNA polymerase1U,10×Buffer2.5μL.23ISSR primers selected from the Shanghai Biological Engineering Company195random primers.393polymorphic fragments obtained for all,319were polymorphic, accounting for81.2%,17fragments for each primer. Based on ISSR analysis of23primers of Lespedeza davurica, genetic similarity among populations of the GS range is0.65to0.89, with anaverage of0.76. According to their genetic similarity coefficient system clustering tree diagram, when GS=0.70to0.72,23populations of Lespedeza davurica were divided into two classes:the populations of2to23were divided into one, population1falled into the second.3. The establishment of Lespedeza davurica AP-PCR system:volume of AP-PCR amplification reaction system is25μL, DNA template60ng, Mg2+1.75mmol/L(final concentration), dNTP0.4mmol/L, primer0.4μmol/L, Taq DNA polymerase1U, lOxBuffer2.5μL.24AP-PCR primers selected from the Shanghai Biological Engineering Company195random primers.333polymorphic fragments obtained for all,301were polymorphic, accounting for90.4%,14fragments for one primer. Based on AP-PCR analysis of24primers of lespedeza davurica, genetic similarity among populations of the GS range is0.57to0.85,with an average of0.70. According to their genetic similarity coefficient system clustering tree diagram, when GS=0.57to0.85,23populations of Lespedeza davurica were divided into two types:the populations of2to23were divided into a type, population1falled into another.Conclusion:the23Lespedeza davurica populations belong to the same group. Genetic distance between population1and22others is0.01to0.03. Because the overall area of Shanxi Province is small(only1.6%of total land area), at the same time, the climate is alike, natural mating between the populations under man-made and natural conditions and so on, so that differences between23populations is little, so23Lespedeza davurica in Shanxi Province were divided into the same population.