Analytical Study Of Angelica Sinensis (Oliv.) Diels Of Gansu Province With RAPD

The genomic DNAs of 20 samples of A. sinensis collected from different regious were isolated and analysed by RAPD. Based on the polymorphism and cluster analysis of their RAPD bands, the molecular phylogenetic tree of 20 A. sinensis samples was described. The results would offer the technological guarantee and molecular biology basis for Chinese angelica resources collection and analysis, origin and evolution of species, classification research, biodiversity protection, genetic map construction and gene location. The result of study was as follows: 1. In the experiment, seven essential factors which may affect the result of RAPD were optimized by orthogonal experiment and two single factor experiments of annealing temperature and cycle times. The result showed that the optimal conditions of RAPD-PCR system and reaction parameters were 0.5 ng/μl template DNA, 0.8 μmol/L primer, 0.25 mmol/L dNTPs, 2.5 mmol/L MgCl2, 0.8 U Taq polymerase.The reaction procedure was 94℃in 3 min, then 40 cycle times with 94℃in 30 s, 36℃in 60 s, 72℃in 90 s, lastly 72℃in 5 min, and stop reaction at 4℃. 2. There were 40 random primers (10-mers) used for the PCR amplification of genomic DNA fragment. Seven of the primers could amplify polymorphic bands, 58 of 82 amplified bands were polymorphic, accounts for 70.7% . Every primer could amplify 5 to 11 polymorphic bands, with an average of 8.3 bands. The value of the similarity matrix of 20 accessions were 0.375 to 1.000 . It showed that the larger its number value is, the nearer the hereditary distance could be. Cluster analysis showed that the samples could be classified into 5 clusters. The result indicated that the smaller the geographic distance between two A. sinensis samples, the smaller their genetic difference, while the genetic difference became greater as their geographic distance increased. However, it should not be ignored that the natural environments, especially the local ecological conditions may also show some influence on the genetic variations among different A. sinensis samples. It is obvious that the result of cluster analysis based on RAPD of A. sinensis are more coincide with geographical distribution of Chinese angelica of Gansu Province.