The Genetic Diversity And Genetic Structure Of Angelica Sinensis From Different Habitats In Gansu Province

Objective: The genetic diversity and genetic structure between different habitats ofAngelica sinensis (Oliv.) Diels was studied by establishing suitable ISSR-PCR reactionsystem for A. sinensis.Methods: The influenced factors of ISSR-RCR reaction were optimized throughorthogonal design and single-factor design. The genetic diversity and genetic structure of A.sinensis from41different habitats in Gansu were studied by optimized system. Analyszingthe different habitats of A. sinensis by Shannon’s diversity information by Popgen1.32software, construct the dendrogram according to Nei’s genetic diversity with UPGMA,estimate the genetic diversity and genetic structure with the statistical data.Results: The optimized system was20μL reaction system containing2.0mmol/L10×PCR Buffer (Mg2+Plus),1.5U Taq DNA polymerase,0.25mmol/L dNTPs,0.4μmol/Lprimers and30ng Template DNA. A total of6ISSR primers from70primers were amplifiedon804samples of A. sinensis from different habitats in Gansu, which had lots of amplifiedbands, strong signal and clearly background.54loci were amplified, in which46werepolymorphic, the average percentage of polymorphic bands (PPB) was85.19%, the Shannon’sinformation index (I) was0.4436and the Nei’s genetic diversity (H) was0.2999. Analysedwith Popgen, Ht of cultivated populations of A. sinensis was0.3012, Hs was0.0204,according to which Gst was calculated as0.9322. Gene flow among populations Estimatedvalue Nm=0.0364. The analysis of Popgen1.32showed that the genetic distances of804samples of A. sinensis from different habitats in Gansu were from0.3994to1.0000. Thegenetic relationships of different samples were analyzed by the dendrogram according to Nei’sgenetic diversity with UPGMA.Conclusion: The established ISSR reaction system would be used to study moleculargenetics of A. sinensis. Each index showed a higher genetic diversity in Gansu cultivated A.sinensis at the species level, but low or flat compared with most other cultivated medicinalherbs, and in the level of genetic diversity within a population was significantly lower thanthat among populations; the genetic differentiation among populations of Gansu cultivated A.sinensis was obvious, and there was no gene exchange among different populations.