Study On Genetic Relationships Among The Part Of Ornamental Bamboos In Phyllostachys Using Random Amplified Polymorphic DNA(RAPD)

Both intraspecific and interspecific relationships of the 23 ornamental bamboos of Phyllostachys in the section were analyzed using RAPD(Randomly Amplified Polymorphic DNA)markers. The species as follows: P. aureaCarr. ex A. et c. riviere two cultivars species of P. sulphurea(Carr.)A.et C. Riv. in Bull. Soc. Acclim cv. Houzeau and cv. Robert young P. arcana f. luteosulcata C. D. Chu et C. S. Cha, P. glauca f. youzhu J. L. Lu, three cultivars species of P. vivax McClure: cv. Vivax McClure, cv. Huanwenzhu and cv. Aureocaulis, two cultivars species of P. praecox CD. Chu et C.S.Chao: cv. Praecox and cv. Notata, four cultivars species of P. heterocycla (Carr ) Mitford: cv. ffeterocycla, cv. Tao Kiang, cv. Obliquinoda and cv. luteosulcata, four cultivars species of P. aureosulcata McClure: cv. Aureosulcata, cv.Pekinensis, cv. spectabiiis and cv.Aureocaulia, two varieties species of P. nigra(Lodd. )Munro. var. nigra and var. Henonis(Mitford)Stapf ex Rendle, P. bambusoides f. lacrimadeae Keng f. et We and P. yunhoensis S. Y. Chen et C. Y. Yao. Six out 27 random primers were selected which producted distinct bands, showing unmistakable polymorphism. By NTSYS-PC program, Jaccard's genetic similarity coefficients were generated and dendrogram was constructed using UMPGA(Unweighted pair-group matheraatic analysis). It is concluded as follows: (1) Amplification reaction system(20uL/tube): 1. OuL 10-mer RAPD primer (50pmol), template DNA 50 ng, 2. OuLlOXreaction buffer(500mmol/L Tris-HCI PH8. 3, 500 mmol/L KCI, 15 mmol/L MgCl2, 0. 01% Gelatin), 1. 0 unit of TaqDNA polymerase, 1. 6uL 25mmol/L dNTPs each.The mixture was joined with super-pure water reach to 20uL.Amplification reaction program: After initial denaturation for 5 min at 94C. Then the reaction was cotinued for another 45 cycles through the followingprocedure: 94C for 45s, 36C for 1 min and 72C for 1 min. Finally, it ended at 72C for 10 min.(2)The polymorphism analysis showed a total of 47 bands, among which 44 were polymorphic, attaining a polymorphic rate of 93. 6%. The number of Base pairs rage of 300 to 1050.Per primer producted 7.8 bands, including 7.3 polymorphic bands. Primer A15 attained the most polymorphic bands among the six primers, polymorphic rate reach 20. 45%.(3)The PCR products using six random primers amplified 23 accessions DNA of Phyllostachys showed the complex genetic base in intraspecific and interspecific and RAPD markers can sensitive detect the genetic diversity. The clustering results according to genetic similarity coefficients and genetic distance were in consistence with that of the analysis of morphology. RAPD markers is not only can detect genetic diversity in interspecific of Phyllostachys and show genetic relationships, but also applied in the systematic study of Phyllos tachys. The conclusion that the variant degree in interspecific relationships of ornamental bamboos is higher than that in intraspecific can help the system evolution research.