Optimization of a SRAP-PCR System for Analysis of Genetic Diversity of Liriodendron
The SRAP-PCR reaction system of Liriodendron was optimized with an orthogonal design to choose the most proper concentrations of Mg2+,dNTPs,primers,Taq polymerase and template DNA at four levels.The concentrations of Mg2+,Taq polymerase,template DNA were further fine-adjusted with a single-factor design.The optimized SRAP-PCR system (total 20 μL)was as follows:Mg2+ 3.0 mmol· L-1,dNTPs 0.3 mmol·L-1,primer 0.9 μmol·L-1,Taq polymerase 2.0 μmol·min-1 and DNA template 90 ng,2 μL 10 × PCR buffer and ddH2O.With the system,genetic diversity of 10 provenances of Liriodendron was analyzed.The results showed that,182 loci were generated from the 15 screened primer pairs,of which 149 loci were polymorphic.The percent of polymorphic loci was 81.87％.The dendrogram of Liriodendron was constructed based on cluster analysis (UPGMA) with the Package of POPGEN 32.The genetic distance and relative relationship of the 10 provenances of Liriodendron was shown by the dendrogram distinctly.It indicated that SRAP molecular marker could be widely used in genetic diversity analysis of Liriodendron.
国家科技期刊平台
NSTL
万方数据
维普
