基于叶绿体基因组SSR分子标记开发鉴定5种生态型天麻  

Identification of five ecotypes of Gastrodia elata based on chloroplast genome SSR molecular marker

作　　者：胡艺镨 李胜男 徐娇 周涛 江维克 欧小宏 HU Yipu;LI Shengnan;XU Jiao;ZHOU Tao;JIANG Weike;OU Xiaohong(Resource Institute for Chinese&Ethnic Materia Medica,Guizhou University of Traditional Chinese Medicine,Guiyang 550025,China;Guizhou Key Laboratory for Germplasm Innovation and Resource-Efficient Utilization of Dao-di Herbs,Guiyang 550025,China)

机构地区：[1]贵州中医药大学中药民族药资源研究院,贵州贵阳550025 [2]贵州省道地药材种质创新与资源高效利用全省重点实验室,贵州贵阳550025

出　　处：《中草药》2025年第5期1747-1754,共8页Chinese Traditional and Herbal Drugs

基　　金：国家重点研发计划(2023YFC3503803);中央本级重大增减支项目(2060302);贵科合学术新苗[2023]-45。

摘　　要：目的建立不同生态型天麻Gastrodia elata叶绿体SSR(chloroplast SSR,cp SSR)分子标记鉴定方法,为天麻分子标记辅助育种提供理论依据。方法利用天麻叶绿体基因组序列获取cpSSR位点,分析比较位点特征并筛选多态性cpSSR,通过聚类分析、主成分分析和遗传结构分析对不同生态型天麻遗传多样性进行分析与鉴定。结果天麻叶绿体基因组中筛选到166个cp SSR位点,其中单核苷酸为主要类型,A/T重复基序占比最高。筛选得到6对cpSSR,共扩增出18个条带,等位基因平均数(number of allele,Na)、有效等位基因数目(effective number of alleles,Ne)、Shannon多样性指数(Shannon index,I)、多态性信息含量指数(polymorphism information content,PIC)分别为3、2.0845、0.5110、0.5110,表明cp SSR多态性较高。通过聚类分析、主成分分析和遗传结构分析均可将不同生态型天麻分为2个类群,其中乌天麻、绿天麻和血红天麻为一类,红天麻和黄天麻为一类。进一步根据CH15和CH202对cpSSR的电泳特异性条带,可以将血红天麻、乌天麻、绿天麻、红天麻和黄天麻进行准确的鉴定。结论利用CH15和CH20两对cpSSR鉴定不同生态型天麻具有效率高、适应性广的优点,对天麻种质鉴定具有重要意义。Objective To establish a chloroplast SSR(cpSSR)molecular marker method for identifying different ecotypes of Gastrodia elata,so as to provide a theoretical foundation for molecular marker-assisted breeding of G.elata.Methods The cpSSR loci were identified from the chloroplast genome sequence of G.elata,followed by the analysis and screening polymorphic cpSSRs.Cluster analysis,principal component,and genetic structure were used to compare and identify the genetic diversity of different ecotypes of G.elata.Results The chloroplast genome of G.elata contained 166 cpSSR loci,with mononucleotide being the most common and A/T repeats motifs being the most frequent.Six pairs cpSSR were selected,which were amplified 18 bands in total.The average number of allele(Na),effective number of allele(Ne),Shannon index(I),and polymorphism information content(PIC)were 3,2.0845,0.5110,and 0.5110,respectively,that indicating high polymorphism in the selected cpSSRs.Genetic diversity analyses,including clustering,principal component analysis,and genetic structure analysis,successfully grouped the G.elata ecotypes into two clusters.One group included the forms of G.elata Bl.f.glauca S.Chow,G.elata Bl.f.viridis Makino,and dark red G.elata Bl.,the other one group included G.elata Bl.f.elata and G.elata Bl.f.flavida S.Chow.Using the cpSSR markers CH15 and CH20,these five G.elata forms could be distinguished by specific bands.Conlusion The cpSSR method utilizing CH15 and CH20 for G.elata ecotype identification is efficient and adaptable,which provides a useful tool for germplasm identification of G.elata.

关 键 词：天麻 生态型 CPSSR 遗传多样性 鉴定 

分 类 号：R286.12[医药卫生—中药学]