淫羊藿属植物PCR-RFLP遗传多样性研究  

PCR-RFLP genetic diversity of plants in Epimedium L.

作　　者：杜明凤[1,2] 李明军 陈庆富[1] 

机构地区：[1]贵州师范大学生命科学学院植物遗传育种研究所,贵州贵阳550001 [2]贵州师范大学研究生处,贵州贵阳550001 [3]贵州省林业调查规划院,贵州贵阳550003

出　　处：《中草药》2012年第3期562-567,共6页Chinese Traditional and Herbal Drugs

基　　金：贵州省中药现代化专项(黔科合农字[2005]5052号)

摘　　要：目的建立淫羊藿植物PCR-RFLP标记系统关系图。方法对主要来自于四川、贵州的17种淫羊藿植物进行了系统的PCR-RFLP标记。结果在8个引物的PCR-RFLP标记分析中,有7个标记能得到1条清晰的谱带;利用12种限制性内切酶对7个标记的扩增产物消化后,在84种引物/酶组合中,共检测到129条酶切片段,其中44条具有多态性;遗传相似系数为0.550～0.988,平均为0.821。结论该属植物遗传关系与其地理分布关系密切。Objective To discriminate Notopterygii Rhizoma et Radix and its adulterants,and secure the quality and clinical curative effect of this medicinal material.Methods The second internal transcribed spacer（ITS2） of ribosomal DNA was amplified and sequenced by bidirectional sequencing of PCR products.Sequence assembly and consensus sequence generation were performed by using CodonCode Aligner.Phylogenetic study was performed using software MEGA 4.0 in accordance with Kimura-2-parameter（K2P） model,and the phylogenetic tree was constructed by using the neighbor-joining（NJ） method.The ITS2 secondary structure was predicted using ITS2 web server.Results The length of ITS2 sequence of Notopterygium incisum and N.franchetii was 228 bp.Their mean intraspecific genetic distance（K2P distance） was far lower than their mean interspecific genetic distance with the adulterants.In the cluster dendrogram,both N.incisum and N.franchetii showed monophyletic,and simultaneously distinguished from their adulterants.To compare the ITS2 secondary structure between the origin plant of Notopterygii Rhizoma et Radix and its adulterants,we noticed that it was obviously distinguished from the adulterants in terms of number,size,position of loop,and the angle of helix exsertion.Conclusion ITS2 barcode could be used to identify Notopterygii Rhizoma et Radix and its adulterants effectively,and then provide important molecular evidence for the authentication of germplasm rescouces and clinic safety of medicinal use.

关 键 词：淫羊藿属 PCR-RFLP 遗传多样性 引物 谱带 

分 类 号：R282.12[医药卫生—中药学]