Abstract:

Researching genetic diversity of Paeonia suffruticosa hybrids is to ensure location of Paeonia suffruticosa hybrids in variety classification system. Paeonia suffruticosa hybrids genomic DNA from leaf tissue was isolated according to the SDS procedure. An orthogonal diagram as experimental design was applied to study the effects of four levels of Mg2+, dNTPs, primers, Taq DNA and genomic DNA polymerase respectively on the reaction system of SRAP, and the SRAP reaction system was optimized by analyzed effects of the concentrations of DNA template. The results showed that all factors had the significant effect on the result of SRAP-PCR. And a better amplification of SRAP-PCR was obtained with the reaction system for Paeonia suffruticosa containing 2.0 mmol/L Mg2+, 1.5 U Taq DNA polymerase, 0.25 mmol/L dNTPs, 2 ng/μL genomic templates DNA and 0.25 μmol/L primer in the total volume of 25 μL. At the same time, 30 primer combinations were selected with the optimized system among 100 primer combinations, which had abundant polymorphism bands. This study would be conducted to the optimized SRAP-PCR system and selection of primers which would play an important role in genetic diversity and genetic relationship in taxonomic hierarchies of Paeonia suffruticosa.