基于三代测序技术对中药灵芝和食用菌rRNA基因及其基因间序列的鉴定研究  

Identification and analysis of ribosomal RNA gene and its intergene(ITSs) in Ganoderma and edible fungus based on third-generation sequencing technology

作　　者：张韦唯 袁育红 戈雅倩 朱彦蓉 赵解南 孙建鹤 黄林佳 胡泽华 许嘉豪 郭琳婷 高宇峰 张益辉 王燕萍 黄钢 王毅兴[6] 戴方平 ZHANG Weiwei;YUAN Yuhong;GE Yaqian;ZHU Yanrong;ZHAO Jienan;SUN Jianhe;HUANG Linjia;HU Zehua;XU Jiahao;GUO Liting;GAO Yufeng;ZHANG Yihui;WANG Yanping;HUANG Gang;WANG Yixing;DAI Fangping(Jiangsu Zhongguo Gene,Nantong 226126,Jiangsu,China;Haimen Hospital,Nantong University,Nantong 226121,Jiangsu,China;Anhui University of Chinese Medicine,Hefei 230012,Anhui,China;Jiangxi University of Chinese Medicine,Nanchang 330004,Nanchang,China;Shanghai Health Medical College,Shanghai 200237,China;Oriental Hospital,Tongji University,Shanghai 200120,China;Higher Engineering College of Pharmacy,Yangtze River Delta,Nantong 226126,Jiangsu,China)

机构地区：[1]江苏中方基因生物医学科技有限公司,江苏南通226126 [2]南通大学附属海门医院,江苏南通226121 [3]安徽中医药大学,安徽合肥230012 [4]江西中医药大学,江西南昌330004 [5]上海健康医学院,上海200237 [6]同济大学附属东方医院,上海200120 [7]长三角药学高等工程学院,江苏南通226126

出　　处：《上海中医药大学学报》2022年第6期5-12,共8页Academic Journal of Shanghai University of Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(81830052);国家重点研发计划项目(2020YFA0909000);上海市分子影像重点实验室建设项目(18DZ2260400);上海市浦东新区中医治未病高峰学科建设项目(PDZY-2018-0603);上海市浦东新区学科带头人项目(PWRd2019-04);江苏省南通市科技局科技基金项目。

摘　　要：目的:基于三代PacBio测序技术,建立中药灵芝和食用菌rRNA 18S-ITS-28S全长序列文库,从分子水平快速、准确鉴定和研究品种。方法:对收集到的8种食用菌和1种中药材真菌灵芝抽提基因组DNA,设计引物扩增18S-ITS-28S全长序列,18S-ITS-28S全长序列包括连续且完整的18S rRNA-ITS1-5.8S rRNA-ITS2和28S rRNA部分序列,其总长度>3.3 kb。将18S-ITS-28S全长序列进行建库和三代PacBio测序。结果:食用菌和药用灵芝的18S rRNA、5.8S rRNA及部分28S rRNA基因序列相似度高且序列保守,而rRNA基因间(ITS1和ITS2)的序列差异性大。结论:本研究的结果提示,分析rRNA基因间(ITSs)区域,而不是仅仅分析rRNA基因,有助于真菌类中药材的鉴定和研究。Objective:Based on the third-generation PacBio sequencing technology,to establish a full-length sequence database of Ganoderma and edible fungi rRNA 18S-ITS-28S,and to identify species quickly and accurately at the molecular level.Methods:DNA was extracted from 8 kinds of edible fungi and 1 kind of Ganoderma lucidum which previously collected,and used primers to amplify the fulllength sequence of 18S-ITS-28S. The full-length sequence of 18S-ITS-28S included continuous and complete 18S rRNA,5.8S rRNA and partial 28S rRNA sequence,the total length is longer than 3.3 kb,the PacBio third-generation sequenceing was used to measure the fulllength sequence of each sample 18S-ITS-28S samples and construct library.Results:The 18S rRNA,5.8S rRNA,and partial 28S rRNA genes of edible fungi and medicinal Ganoderma lucidum were highly similar and conserved. The rRNA genes(ITS1 and ITS2)were significantly different.Conclusion:The results of this study suggest that the analysis of rRNA intergenic(ITSs)regions,instead of only the analysis of rRNA genes,is helpful for the identification and research of fungal Chinese medicinal materials.

关 键 词：灵芝 食用菌 RRNA基因 测序分析 鉴定 

分 类 号：R282.5[医药卫生—中药学]