Ge, The Case Of Rapd Analysis

RAPD technique was first used to analyze the phylogenetic relationships of 13 species or varieties in Pueraria DC complex. Seventy-two random primers were tested,among which 18 chosen primers generated clear polymorphic bands. Fragments generated by the primers ranged in size from 300 bp to approximately 3 kb. A total of 171 bands were amplified ranging from 1 to 16 bands each primer,151(88%) of which were polymorphic bands. The genetic distances between all pairs of test materials were calculated and phylogenetic relationships among them were discussed.At distance of 0.53,13 materials were divided into 5 groups distinctly:(I) six materials (Material No.l,3,12,13,7 and 2),(II) three materials of the same district (Material No.8,9 and 10),(III) two materials of P.lobata (Material No.5 and 6),(IV) material 4 and (V) sole material 11 of P. Montana. Furthermore group I and II can be divided into subgroups. Material 1 and 3 are P. lobata (wild) from different states of the USA. Material 12 and 13 are P.lobata originating from the same districts of Anhui Province,China. Material 5 and 6 are also P. lobata originating from the same regions of Shaanxi Province,which were closely related to each other,so they were first clustered into the same group or subgroup respectively. Material 8,9 and 10 were all from Guangdong Province,China,and were clustered together. Material 8 and 9 belong to the same species (P. thomsonii),so at smaller genetic distance they were clustered into the same subgroup. Material 11 was the only species P. montana . So it was distantly related to other materials and formed independent group.In contrast to our expectation,material 4 wasn't clustered into the group of other P. lobata from the USA and material 7 wasn't clustered into the group of materials from Shaanxi,China. Material 4 wasintroduced from America and was cultivated in Anhui Province,China,which was distantly related to the same species originating from Anhui Province. To a certain degree,we consider it was affected greatly by Anhui local ecological environments and its genotype was changed greatly. Material 7 was P. lobata originating from Shaanxi province,which was clustered into the same group of P. lobata from other regions. Therefore they probably originated from the same location dating back to times.The results showed that the genetic diversity was not only among species or varieties but also regions. Outer environmental conditions have much effect on the genotype of Pueraria DC. The complicated genetic variation in genus Pueraria is related to geographic distribution.