Study On Rhododendron Using Random Amplified Polymorphic DNA Analysis (RAPD)

The reaction Condiactions for the Randomly Amplified Polymorphic DNA (RAPD) analysis for Rhododendron have been optimized and analyzed the Genetic Relationships Among 24 Rhododendron using RAPD markers. Four out 60 random primers were selected which producted distinct bands, showing unmistakable polymorphism. By NTSY-PC program, Jaccard's genetic similarity coefficients were generated and dendromgram was constructed using UMPGA. The results were as fellow:1)The PCR products were stable when the PCR was 45 cycles of predenature at 95℃ for 1 minutes, denature 94℃ 1 minutes, annealing 34℃ 1 minutes, extension 72℃ 2minutes. Finally, it extenst 72℃ 5 minutes, and stored at 4℃.The amplification solution(20ul) was consisted of dNTPs 0.6 ul, 10×PCR buffer2 μl, MgCl₂ ( 25mmol/L ) 1.8μl, Taq DNA polymerase ( 0.5U/μl ) 0.3μl,Primer(0.8Pm/μl)4μl,template DNA (10ng/μl) 1.5μl.2) The polymorphism analysis showed a total of 48bands ,among which 44were polymorphic.Per primer producted 12 bands, including 11 polymorphic bands. Primer U10 attained the most polymorphic bands among the four primers, polymorphicrate reach 93.33%.3)The PCR products using the four random primers amplified 24 accession DNA of Rhododendron showed the complex genetic base in the Genetic Relationships. The clustering results according to genetic similarity coefficients and genetic distance were in consistence with that of the analysis of morphology. RAPD is not only can detect genetic diversity in Rhododendron and show genetic relationships, but also applied in the systematic study of Rhododendron.