溪黄草药材基原植物及近缘植物ISSR-SCAR分子标记开发  

Development of ISSR-SCAR Molecular Markers for Original Plants and Related Plants of Rabdosia Herba

作　　者：刘靖 孟爽爽 刘军民[1,2] 萧晓吉 古敬锋 龚彩玲 谢文波 赵双双 詹若挺 LIU Jing;MENG Shuangshuang;LIU Junming;XIAO Xiaoji;GU Jingfeng;GONG Cailing;XIE Wenbo;ZHAO Shuangshuang;ZHAN Ruoting(School of Chinese Materia Medica,Guangzhou University of Chinese Medicine,Guangzhou 510006,China;Key Laboratory of Chinese Medicinal Resource from Lingnan(Guangzhou University of Chinese Medicine),Ministry of Education,Guangzhou 510006,China;Department of Pharmacy,Shenzhen Hospital of Traditional Chinese Medicine,Shenzhen 518033,China;Guangdong Yunfu Vocational College of Chinese Medicine,Yunfu 527400,China;Guangzhou Pharmaceutical Vocational School,Guangzhou 510430,China;China Resources Sanjiu Medical&Pharmaceutical Co.,Ltd.,Shenzhen 518110,China;Guangdong Agribusiness Tropical Agriculture Research Institute Co.,Ltd.,Guangzhou 511365,China)

机构地区：[1]广州中医药大学中药学院,广州510000 [2]岭南中药资源教育部重点实验室(广州中医药大学),广州510006 [3]深圳市中医院药学部,广东深圳518033 [4]广东云浮中医药职业学院中药学院,广东云浮527400 [5]广州市医药职业学校,广州510430 [6]华润三九医药股份有限公司,广东深圳518002 [7]广东农垦热带农业研究院有限公司,广州511365

出　　处：《中国药学杂志》2024年第15期1400-1407,共8页Chinese Pharmaceutical Journal

基　　金：2022年省级乡村振兴战略专项资金种业振兴项目资助(2022-NJS-00-002);云浮中医药(南药)产业创新团队项目资助(云科函[2023]96号,202301);广东省教育厅2021年广东省本科高校教学质量与教学工程改革建设项目资助(粤教高函[2021]29号)。

摘　　要：目的研究建立溪黄草药材基原植物及其近缘植物的特征性片段扩增区域(SCAR)分子标记技术,为溪黄草药材的基原鉴定提供新的分子鉴别手段。方法采集溪黄草药材的基原植物及其近缘植物共5种29个居群,采用简单序列重复区间扩增多态性(ISSR)分子标记对代表居群的基因组DNA进行聚合酶链式反应(PCR)扩增,获得ISSR特异性片段,经回收纯化、克隆、测序后,根据特异性片段测序结果设计可以直接用于它们高效鉴定的SCAR引物。结果采用14对ISSR引物对18个代表样本进行聚合酶链式反应PCR扩增,获得4条特异性片段并成功转化为ISSR-SCAR分子标记,可分别用于特异性识别线纹香茶菜Rabdosia lophanthoides(Buch.-Ham.ex D.Don)H.Hara、长叶香茶菜R.stracheyi(Benth.Ex Hook.f.)Hara、溪黄草R.serra(Maxim.)H.Hara和显脉香茶菜R.nervosa(Hemsl.)C.Y.Wu et H.W.Li。结论建立的ISSR-SCAR分子标记条带清晰明亮,结果稳定,能够准确、快速鉴别溪黄草药材的基原植物及其近缘植物,为溪黄草药材基原的分子鉴别提供了新的方法。OBJECTIVE To study and establish sequence characterized amplifiedregion(SCAR)molecular marker for identifying the original plants and related plants of Rabdosia Herba,and provide a new molecular identification method for the original plants of Rabdosia Herba.METHODS A total of 29 populations of the original plants and related plants of Rabdosia Herba were collected,and inter-simple sequence repeat(ISSR)molecular marker was used to perform PCR amplification on the genomic DNA of the representative population of Rabdosia Herba to obtain ISSR-specific segments.After the ISSR-specific fragments were recovered,purified,cloned and sequenced,SCAR primers were designed based on the sequencing results,which were directly used for the efficient identification of the original plants of Rabdosia Herba.RESULTS Fourteen pairs of ISSR primers were used to perform PCR amplification on 18 representative samples of original plants and related plants of Rabdosia Herba,and 4 specific segments were obtained and successfully transformed into ISSR-SCAR molecular markers.These molecular markers can be used to specifically identify the Rabdosia lophanthoides(Buch.-Ham.ex D.Don)H.Hara,Rabdosia stracheyi(Benth.Ex Hook.f.)Hara,Rabdosia serra(Maxim.)H.Hara,and Rabdosia nervosa(Hemsl.)C.Y.Wu et H.W.Li,respectively.CONCLUSION The established ISSR-SCAR molecular markers produce clear and bright bands with stable results,which can accurately and rapidly identify the original plants and related plants of Rabdosia Herba,providing a new method for the identification of the original plants of Rabdosia Herba.

关 键 词：溪黄草药材基原植物 分子标记 简单序列重复区间扩增多态性 特异片段 

分 类 号：R282[医药卫生—中药学]