Study On Genetic Diversity Of The Ancient Sophora Japonica L. By SRAP Analysis

Sophora japonica L. is a tree species with timber, medicinal, edible and ornamentalvalues.extensive introduction and cultivation in other parts of the world. In order to better forappraisal of Sophora Japonica germplasm resources, this paper analysed genetic diversity anddiscussed the genetic similarity of Sophora Japonica with the method of SRAP marker. It wasexpected that research results would provide the basis for the breeding of new varieties andgermplasm resources.The main results from this study are as follows：1、Genomic DNA was extracted from leaves of by the method of Sophora japonica modifiedCTAB. We obtained high quality and purity genomic DNA which can meet the request ofPCR reaction.2、An orthogonal of L16(45) design combined with single factor experiment was applied tooptimize SRAP-PCR amplification system of Sophora japonica. The effect of the mainreaction system elements on SRAP-PCR were tested. A most suitable SRAP-PCR system forSophora japonica. Was established that is total20μL reaction system containin: Mg2+2.0mmol/L, DNA template80ng, dNTPs0.15mmol/L, primer1μmol/L,10×PCR buffer2.5mmol/L and ExTaq DNA polymerase1.0U. And the optimal annealing temperature was50degrees℃.3、Preservation using laid a foundation6primer pairs were screened out from100SRAPprimer pair combinations based on their stable amplification, clear banding patterns and goodpolymorphism by utilizing the SRAP reaction system. each primer combination produced4-11poly-morphic loci; Percent of Polylnorphic loci54.16%-80.55%with an average of65.4%.4、Sophora japonica provenance of genetic diversity index:Total number of allels (Na) was1.8889of, effective number(Ne) was1.6527, Nei’s gene diversity (H) was0.3691, Shannon’sInformation index (I) was0.5359，It was basically identical that genetic level in32provenanceeffective number of each index. The significance difference did not exist in genetic diversityof32provenance by analysis.5、Genetic diversity index of provenances of Sophora japonica:Ht=0.3691, Hs=0.3338,Gst=0.0958, The percentage of genetic diversity among the provenances was9.58%in the total value, The percentage of genetic diversity within provenances was90.43%in the totalvalue, that means the genetic variation was mainly occurred inner provenances. Nm=4.7178,The mean of gene flow(Nm) was4.7178, which showed relatively steady gene flow amongprovenances.6、Sophora japonica genetic distance among32materials was ranged from0.1040to0.9163,with an average of0.3979; The genetic similarity among32materials was rangedfrom0.3333to0.8889. with an average of0.6190.description of the genetic relationshipbetween sophore japonica provenance relatively close.7、Base on the UPGMA cluster diagram, the32materials of Sophora japonica may be dividedwith the genetic distance of0.41into7groups.The result was not completely consistent withthe traditional division of provennans. That means geogrclaphic location does not match thedescription of clustering results with a variety of sources.8、In this study, the highest correlation value between typical Variables V1and plant heightwes0.8955, and the correlation coefficients between Typical variable W1and leaf width,rachis spacing were0.6389and0.6389, respectively, illustrated that this Typical variableMain reaction there is larger negative correlation relationship between plant height and leafwidth and rachis spacing, mean that the higher plant, the relatively larger leaf width and rachisspacing.