WANG Xia,XU Jize,WANG Huan,et al.Establishment of ISSR Reaction System and Genetic Diversity Analysis of Hypericum attenuatum Choisy.[J].Northern Horticulture,2017,41(05):96-101.[doi:10.11937/bfyy.201705023]
乌腺金丝桃ISSR反应体系的构建及其遗传多样性分析
- Title:
- Establishment of ISSR Reaction System and Genetic Diversity Analysis of Hypericum attenuatum Choisy.
- 文献标志码:
- A
- 摘要:
- 以14份乌腺金丝桃为试材，采用L16（45）正交实验设计，建立并优化乌腺金丝桃的ISSR-PCR反应体系，并利用优化的反应体系分析供试材料间的遗传多样性。结果表明：20 μL ISSR-PCR反应体系中应含有Taq DNA聚合酶1.0 U，dNTPs 0.2 mmol?L-1，Mg2+ 2.0 mmol?L-1，模板DNA 45 ng。从45条ISSR引物中筛选出11条多态性引物，利用筛选的多态引物对14份乌腺金丝桃材料进行了ISSR遗传多样性分析，共扩增出85条带，其中多态性条带57条，占67.06%;4个乌腺金丝桃居群的遗传相似系数介于0.375 1～0.725 2，平均值为0.541 6;通过UPGMA进行聚类分析表明，14份材料可分为3个类群4个亚群。乌腺金丝桃居群间遗传多样性水平明显高于居群内，其遗传距离与地理距离具有一定的相关性。
- Abstract:
- Fourteen Hypericum attenuatum were used as materials,the ISSR-PCR reaction system was established and optimized by L16(45) orthogonal experimental design.The genetic diversity of Hypericum attenuatum was analyzed by the optimal reaction system.The results showed that the optimum reaction system contained 1.0 U Taq DNA polymerase，0.2 mmol?L-1 dNTs，2.0 mmol?L-1 Mg2+ and 45 ng DNA template respectively in a 20 μL ISSR reaction system.Eleven polymorphic primers which were used to analyze the genetic diversity of Hypericum attenuatum were selected from 45 ISSR primers.A total of 85 bands were amplified，of which 57 bands were polymorphic loci and the percentage of total polymorphic loci was 67.06%.The genetic similarity coefficient was 0.375 1-0.725 2 in 4 populations of Hypericum attenuatum,and the average was 0.541 6.The clustering analysis was constructed by UPGMA method based on the genetic identity and 14 Hypericum attenuatum materials were clustered into 3 main groups and 4 subgroups.The level of genetic diversity of Hypericum attenuatum between populations was higher than that within populations and between the genetic distance and geographic distance had a certain correlation.
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备注/Memo
第一作者简介：王霞(1978-)，女，硕士，讲师，研究方向为植物分子生物学及天然产物分离。E-mail:wangxhangj@126.com.基金项目：长白山重点实验室培育资助项目(吉农院合字[2014]第s004号）。