Establishment Of RAPD, ISSR, And SSR Reaction System And Analysis Of Genetic Diversity Of Japanese Plum Cultivars

Japanese plum (Prunus salicina ) is one of important fruit tree among China and world. In this paper, RAPD, ISSR, and SSR reaction system of the fruit tree were established, and the genetic diversity of 56 genotypes of Japanese plum were analyzed using RAPD, ISSR, and SSR markers. The optimal reaction of RAPD was studied with 5'-GACCGCTTGT-3' primer for cv. Fenghuali. The results showed that the optimum concentration of five important components i.e. Tag DNA polymerase, Mg2+, primer, template DNA and dNTPs in 25μL RAPD reaction system were 1.0u, 2.5mM, 20ng, 40ng and 0.2mM, respectively. The RAPD fingerprinting of five Japanese plum cultivars were established by using optimum reaction system with ten random primers. The genetic relationship of these cultivars were analyzed with their Band Sharing (BS). The Band Sharing average of five plum cultivars was 33.2%, and their variant range was 20.1%~40.9%. The optimal reaction of ISSR in Prunus salicina cv. Niuxinli was studied with primer 5'-(AC)9A-3'. The results showed that the optimum concentration of five important components i.e. Taq DNA polymerase, Mg2+, primer, template DNA and dNTPs in 25 u L reaction system of ISSR were 0.3u, 1.5mM, 0.2 μM, 20ng and 0.16mM, respectively. The ISSR fingerprinting of 18 Japanese plum cultivars and 12 plum species or variety were established by using optimum system with primer 5'-(AC)9T-3'and 5'-(AC)9C-3', respectively. The identification rate of plum cultivars and species or variety was 100%. The effects of five important components on reaction of SSR were studied with Prunus salicina cv. Meilili. The results showed that the optimum concentration of five important components i.e. Tag DNA polymerase, Mg +, single primer, template DNA and dNTPs in 25 μL reaction system of SSR were 1.5u, 2.0mM, 0.8 μ M, 30~40ng and 0.16~0.24mM, respectively. The dendrograms of these cultivars was contructed according the Jaccard coefficient of similarity. The results were showed that a high genetic diversity was observed using these molecular markers. Fifty-four Japanese plum cultivars were classified five groups from the dendrogram. The majority of cultivars of northern areas in China were clustered together, and cultivars of southern areas in China were clustered together. Therefore, the Japanese plum cultivars in China could be classified two groups: northern cultivars group (NCG) and southern cultivars group (SCG).