不同品种怀地黄药用部位EST-SSR遗传特性研究  

Study on EST-SSR Genetic Characteristics of Medicinal Parts of Different Varieties of Rehmannia glutinosa

作　　者：宋梦娇 董诚明[1,3,4] 魏悦 杨林林 初雷霞 张岩岩 侯文川 SONG Meng-jiao;DONG Cheng-ming;WEI Yue;YANG Lin-lin;CHU Lei-xia;ZHANG Yan-yan;HOU Wen-chuan(Henan University of Chinese Medicine,Zhengzhou 450046,China;Henan Natural Product Biotechnology co.Ltd,Zhengzhou 450002,China;Henan Provincial Ecological Planting Engineering Technology Research Center of Authentic Medicinal Materials,Zhengzhou 450046,China;Henan University of Chinese Medicine,Provincial and Ministry Co-construction of Collaborative Innovation Center about Prevention and Treatment of Respiratory Diseases in Traditional Chinese Medicine,Zhengzhou 450046,China)

机构地区：[1]河南中医药大学药学院,河南郑州450046 [2]河南省纳普生物技术有限公司,河南郑州450002 [3]河南省道地药材生态种植工程技术研究中心,河南郑州450046 [4]河南中医药大学呼吸疾病中医药防治省部共建协同创新中心,河南郑州450046

出　　处：《时珍国医国药》2023年第9期2144-2147,共4页Lishizhen Medicine and Materia Medica Research

基　　金：国家药品监督管理局中药材及饮片质量控制重点实验室开放课题(KF202202);国家重点研发计划(2017YFC1702800);河南省科学院基本科研项目(210913009,220613105,230613075)。

摘　　要：目的采用EST-SSR分子标记法,对怀地黄新品种“怀中1号”和道地产区4个主流品种药用部位进行种间遗传特性研究。方法优化DNA提取方法提取地黄药用部位DNA,筛选地黄SSR-PCR引物,采用Popgene32软件计算遗传距离和遗传相似系数进行分析,Ntsyspc2.1对地黄扩增结果进行聚类分析。结果从68对引物中筛选出5对具有多态性的引物,共扩增出97条条带,其中多态性条带78条,多态性条带比率为80.41%,每对引物检测到的等位基因数为10~21个,平均15.6个。有效等位基因为1.1697~1.3332,Nei’s基因多样性为0.0980~0.1980,Shannon指数为0.1461~0.3023,不同品种的遗传距离范围为0.0665~0.1523。聚类分析结果显示,遗传相似系数为0.74时,5个品种分为3类,脱毒北京3号和怀丰聚为一类,北京3号和金九聚为一类,怀中1号为一类。结论EST-SSR分子标记法可以用于不同品种地黄药用部位间的亲缘关系鉴定,为地黄遗传稳定性研究及品种鉴定提供科学研究依据。Objective The interspecific genetic characteristics of“Huaizhong No.1”a new variety of Rehmannia glutinosa and 4 mainstream varieties of medicinal parts in genuine producing area were studied by using the EST-SSR molecular marker method.Methods Optimizing the DNA extraction method to extract the DNA from the medicinal parts of R.glutinosa,the SSR-PCR primers of R.glutinosa were selected,Popgene32 software was used to calculate the genetic distance and genetic similarity coeffi-cient for analysis,and Ntsyspc2.1 was used to perform cluster analysis on the amplification results of R.glutinosa.Results The results showed that 5 pairs of primers with polymorphism were selected from 68 pairs of primers.A total of 97 bands were ampli-fied,including 78 polymorphic bands,the ratio of polymorphic bands was 80.41%.And the number of alleles detected by each pair of primers ranged from 10 to 21,with an average of 15.6.Effective alleles were 1.1697~1.3332,Nei’s gene diversity was 0.0980~0.1980,Shannon index was 0.1461~0.3023,and the genetic distance of different breeds were ranged from 0.0665 to 0.1523.The cluster analysis results showed that when the coefficient of genetic similarity was 0.74,the 5 varieties were divided into 3 groups,Detoxification Beijing No.3 and Huaifeng were grouped together,Beijing No.3 and Jinjiu were grouped together,and Huaizhong No.1 was grouped separately.Conclusion The EST-SSR molecular marker method can be used to identify the genetic relationship between the medicinal parts of different varieties of R.glutinosa,and provide a scientific research basis for the study of genetic stability of R.glutinosa and variety identification.

关 键 词：怀地黄 EST-SSR 遗传稳定性 品种鉴定 

分 类 号：S567[农业科学—中草药栽培]