不同地域的冬虫夏草rDNA ITS区的序列分析和遗传分化  

Analyses and Genetic Variation of rDNA ITS in Ophiocordyceps sinensis from Different Geographical Regions

作　　者：汪家春[1] 徐军[2] 李兆兰[3] 王玥[4] 陈雅琳[1] WANG Jia-chun;XU Jun;LI Zhao-lan;WANG Yue;CHEN Ya-lin(PLA Navy Medical research institute,Shanghai 200433,China;Shanghai Health Medical College,Shanghai 200093,China;College of Life Science,Nanjing University,Nanjing 210093,China;College of Life Science and Technology,Xinjiang University,Wulumuqi,830046,China)

机构地区：[1]中国人民解放军海军医学研究所,上海200433 [2]上海健康医学院,上海200093 [3]南京大学生命科学学院,南京210093 [4]新疆大学生命科学与技术学院,乌鲁木齐830046

出　　处：《天然产物研究与开发》2018年第11期1963-1970,共8页Natural Product Research and Development

基　　金：解放军总后勤部科研项目(11DJZ2b);全军医学科技青年培训项目(13QPO72);上海教育委员会科研创新项目(11YZ293)

摘　　要：了解不同地区的冬虫夏草种内的遗传分化及变异情况,进行18S rDNA ITS区的序列分析,以期为它们的差别建立分子标记,也为不同地区来源的商品冬虫夏草的鉴定提供分子依据。冬虫夏草样品应用DNA提取试剂盒所提供的方法(CTAB法)提取基因组DNA,1%琼脂糖凝胶电泳检测DNA纯度,DNA经PCR扩增,PCR扩增产物在DNA全自动测序仪上进行序列测定。测序结果获得6个不同地域的冬虫夏草18S rDNA ITS区基因序列图谱和GC含量(%)。实验结果表明,6个不同地区的冬虫夏草基因序列,反映了冬虫夏草种内的遗传分化和变异,其遗传分化与变异的程度与海拔高度和纬度呈正相关性,这些分化可应用于不同地区冬虫夏草的鉴定。To investigate the genetic variation among Ophiocordyceps sinensis from different geographical regions,the sequence of 18S rDNA ITS was analysed in Ophiocordyceps sinensis,in order to establish the molecular genetic marker for O.sinensis from different geographical regions.Total DNA of dry specimens was extracted from stroma or sclerotium powder of O.sinensis using a modification of the DNA isolations protocol of Lee&Taylor(1990).The crude extracts were examined by 1%agarose gel electrophoresis in 1×TAE buffer.The extracts were used as templates and were amplified with PCR technology,PCR amplification products were purified,sequenced and compared.The present study showed that sequences of 18S rDNA ITS there were obvious genetic variations and dissimilarity among in O.sinensis from different geographical regions,such as,sequences composition,GC content(%)etc.The results showed that genetic differentiation of O.sinensis has close relation to latitudes and elevation highness.These differences can be used to differentiate locality identificiation of O.sinensis.

关 键 词：冬虫夏草 rDNAITS PCR扩增 遗传分化和变异 海拔高度和纬度 

分 类 号：R282.5[医药卫生—中药学] Q939.5[医药卫生—中医学]