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通过正交试验设计,比较苦荞序列相关多态性-聚合酶链式反应(Sequence related amplified polymorphism-polymerase chain reaction,SRAP-PCR)扩增反应体系的组合,选出最佳扩增体系用于苦荞SRAP分子标记进行遗传多样性分析。其中最佳SRAP-PCR反应扩增体系为Taq DNA聚合酶2 U、Mg2+1.5 mmol/L、DNA模板75ng、引物0.5μmol/L。从238对SRAP引物组合中筛选出20对多态性较高的引物组合,并对15个苦荞品种进行遗传多样性分析。结果表明:20对引物组合共扩增出128个位点,其多态性信息量(Polymorphism information content,PIC)值在0.66~0.95,每对引物组合扩增位点为7~13个,多态性比率平均值为81.6%,其中多态性信息量值为0.95的引物组合为me7em11、me9em4和me12em8。基于UPGMA法聚类(GS=0.78)可将15个苦荞品种划分为3大类,但这3类划分的品种没有明显的地域分布特征。
Through orthogonal experimental design, we compared the combination of amplification reaction system of sequence related amplified polymorphism-polymerase chain reaction (SRAP-PCR) and selected the best amplification system Genetic Diversity Analysis of SRAP Molecular Marker of Tartary Buckwheat. Among them, Taq DNA polymerase 2 U, Mg2 + 1.5 mmol / L, DNA template 75 ng and primer 0.5 μmol / L were the best amplification system for SRAP-PCR reaction. Twenty pairs of primer combinations with high polymorphism were screened from 238 pairs of SRAP primer combinations and genetic diversity of 15 tartary buckwheat varieties was analyzed. The results showed that a total of 128 loci were amplified by 20 pairs of primers. The polymorphism information content (PIC) values ranged from 0.66 to 0.95, and the number of amplified regions per primer pair ranged from 7 to 13 The average ratio of polymorphic loci was 81.6%, and the primer combinations with polymorphic informative value of 0.95 were me7em11, me9em4 and me12em8. Based on UPGMA clustering (GS = 0.78), 15 tartary buckwheat cultivars could be divided into three groups, but the three cultivars had no obvious geographical distribution.