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ISSR-PCR反应体系为:总体积20μL,包含10×PCR Buffer 2μL、dNTPs 0.5 mmol/L、Taq酶1.0 U、MgCl 23.5 mmol/L、引物0.25μmol/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0.4921~0.7302之间,遗传距离在0.3145~0.7091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。
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 pair-group method arithmetic averages(UPGMA).The ISSR-PCR reaction system in this experiment(20μL)contained 10×PCR Buffer 2.0μL、0.5 mmol/L dNTPs、1.0 U Taq DNA polymerase、3.5 mmol/L MgCl 2、0.25μmol/L primer、and 50 ng template DNA.A total of 118 polymorphic bands were amplified by 14 primers.Genetic consistency ranged from 0.4921 to 0.7302 and the genetic distance ranged from 0.3145 to 0.7091.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.
作者
崔洁
王丹丹
王倩玉
陈彤垚
张福生
秦雪梅
CUI Jie;WANG Dan-dan;WANG Qian-yu;CHEN Tong-yao;ZHANG Fu-sheng;QIN Xue-mei(Modern Research Center for Traditional Chinese Medicine;College of Chemistry and Chemical Engineering;Key Laboratory of Effective Substances Research and Utilization in TCM of Shanxi Province,Shanxi University,Taiyuan 030006,China;Shijiazhuang No.4 Pharmaceutical.4Co.,Ltd.,Shijiazhuang 050021,China)
出处
《天然产物研究与开发》
CAS
CSCD
北大核心
2019年第11期1941-1948,共8页
Natural Product Research and Development
基金
国家自然科学基金(31770362)
关键词
蒙古黄芪
ISSR体系优化
遗传多样性
种质资源
Astragalus membranaceus
ISSR system optimization
genetic diversity
germplasm resources
作者简介
通信作者:张福生,Tel:86-351-7019178,E-mail:ample1007@163.com;通信作者:秦雪梅,qinxm@sxu.edu.cn。
