山西与内蒙产蒙古黄芪的ISSR体系优化及遗传多样性分析

doi:10.16333/j.10016880.2019.11.013

天然产物研究与开发 ›› 2019, Vol. 31 ›› Issue (11): 1941-1948.doi: 10.16333/j.10016880.2019.11.013

山西与内蒙产蒙古黄芪的ISSR体系优化及遗传多样性分析

崔洁1,2,3，王丹丹1,4，王倩玉1,2,3，陈彤垚1,3，张福生1,3*，秦雪梅1,3*

1山西大学中医药现代研究中心；2山西大学化学化工学院；3山西大学地产中药功效物质研究与利用山西省重点实验室，太原 030006；4石家庄四药有限公司，石家庄 050021

出版日期:2019-11-28 发布日期:2019-12-16
基金资助:
国家自然科学基金（31770362）

ISSR system optimization and genetic diversity analysis of Astragalus membranaceus produced in Shanxi and Inner Mongolia

CUI Jie1,2,3,WANG Dandan1,4,WANG Qianyu1,2,3,CHEN Tongyao1,3,ZHANG Fusheng1,3*,QIN Xuemei1,3*

1Modern Research Center for Traditional Chinese Medicine;2College of Chemistry and Chemical Engineering;3Key Laboratory of Effective Substances Research and Utilization in TCM of Shanxi Province,Shanxi University,Taiyuan 030006,China;4Shijiazhuang No.4 Pharmaceutical.4Co.,Ltd.,Shijiazhuang 050021,China

Online:2019-11-28 Published:2019-12-16

摘要/Abstract

摘要： 本研究对影响蒙古黄芪ISSR反应体系的因素进行了优化，同时对蒙古黄芪的遗传多样性进行了分析。确立蒙古黄芪ISSR初始反应体系，采用单因素实验优化ISSR反应体系，确立了最终的ISSR体系。采用非加权平均距离法（UPGMA）对8批蒙古黄芪样品进行遗传多样性和聚类分析。此次实验获得的黄芪ISSRPCR反应体系为：总体积20 μL，包含10×PCR Buffer 2 μL、dNTPs 0.5 mmol/L、Taq酶1.0 U、MgCl₂ 3.5 mmol/L、引物0.25 μmol/L、DNA 50 ng。14条引物共扩增出多态性位点118个，遗传一致度在0.492 1~0.730 2之间，遗传距离在0.314 5~0.709 1之间。由聚类分析可知8批蒙古黄芪以产地分为了3类，内蒙古兴和县，山西浑源官儿乡各聚为一类，其余山西不同产地聚为一类。本实验表明建立的蒙古黄芪ISSR体系稳定，而山西与内蒙2个产地的蒙古黄芪遗传多样性丰富，该结果为黄芪良种选育及种质保护奠定基础。

关键词: 蒙古黄芪, ISSR体系优化, 遗传多样性, 种质资源

Abstract: In this study,the factors affecting the ISSR reaction system of Astragalus membranaceus were optimized,and the genetic diversity of A. membranaceus was analyzed.The initial ISSR reaction system of A. membranaceus was established,and the ISSR reaction system was optimized by single factor experiment.Then,the ISSR system was finally established.Genetic diversity and cluster analysis of 8 batches of A. membranaceus was analyzed by unweighted pairgroup method arithmetic averages (UPGMA).The ISSRPCR reaction system in this experiment (20 μL) contained 10×PCR Buffer 2.0 μL、0.5 mmol/L dNTPs、1.0 U Taq DNA polymerase、3.5 mmol/L MgCl₂、0.25 μmol/L primer、and 50 ng template DNA.A total of 118 polymorphic bands were amplified by 14 primers.Genetic consistency ranged from 0.492 1 to 0.730 2 and the genetic distance ranged from 0.314 5 to 0.709 1.The results of UPGMA clustering showed that Xinghe County of Inner Mongolia,and Guaner Township of Hunyuan country,Shanxi Province belonged to one class,respectively,while the rest of Shanxi different areas were clustered into other class.This experiment showed that the establishment of the A. membranaceus ISSR system was stable.The genetic diversity of A. membranaceus in two producing areas in Shanxi and Inner Mongolia was rich,which laid a foundation for breeding and germplasm protection of A. membranaceus.

Key words: Astragalus membranaceus, ISSR system optimization, genetic diversity, germplasm resources

中图分类号:

R282.71

崔洁1,2,3，王丹丹1,4，王倩玉1,2,3，陈彤垚1,3，张福生1,3*，秦雪梅1,3*. 山西与内蒙产蒙古黄芪的ISSR体系优化及遗传多样性分析[J]. 天然产物研究与开发, 2019, 31(11): 1941-1948.

CUI Jie1,2,3,WANG Dandan1,4,WANG Qianyu1,2,3,CHEN Tongyao1,3,ZHANG Fusheng1,3*,QIN Xuemei1,3*. ISSR system optimization and genetic diversity analysis of Astragalus membranaceus produced in Shanxi and Inner Mongolia[J]. NATURAL PRODUCT RESEARCH AND DEVELOPMENT, 2019, 31(11): 1941-1948.

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山西与内蒙产蒙古黄芪的ISSR体系优化及遗传多样性分析

ISSR system optimization and genetic diversity analysis of Astragalus membranaceus produced in Shanxi and Inner Mongolia

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