三叶木通SRAP体系的优化及其遗传多样性

Study on optimization of SRAP-PCR reaction system and genetic diversity in Akebia trifoliate

采用L16(45)的正交试验确立了适合三叶木通(Akebia trifoliate)相关序列扩增多态性(SRAP)最佳反应体系为(25μL):1×Reaction Buffer、Mg2+2.5mmol/L、dNTPs0.2mmol/L、Taq酶2U、引物0.4μmol/L、DNA 20ng/25μL.利用最佳体系从100对SRAP引物中筛选出6对,对秦岭地区3个居群的62份三叶木通进行遗传多样性分析,共得到142条条带,多态性比率为87.32%.聚类分析结果显示:在遗传相似系数为0.72时,可将62份三叶木通划分为3个组群,与样品采集地的居群相一致.表明该SRAP体系能够提供丰富的信息位点,适用于三叶木通遗传多样性的分析.

SRAP-PCR amplification system on Akebia trifoliate was optimized by the orthogonal design of L16 （4^5 ）,The optimal PCR system was obtained as 1 × Reaction Buffer,2.5 mmol/L Mg^2＋,0.2 mmol/L dNTPs,2 U TaqDNA polymerase,0.4μmol/L primer,20 ng genomic DNA in 25μL reaction system.6 primer pairs from 100 primer combinations were selected to analyze the genetic diversity relationship of 6 2 strains Akebia trifoliate from Qinling region.There were 142 clear bands produced,the rate of polymorphism was 87.32%.The result of cluster analysis based on SRAP markers showed that 62 stains Akebiatrifoliate could be clustered into 3 groups at 0.72 of genetic similarity coefficient.Every individual of one population was clustered in the same group.It is consistant with the sample collecting zone in group.Therefore,the SRAP system can provide abundant information site,which can be used in the research of genetic diversity of Akebia trifoliate in future.