沪地龙COⅠ条形码和特征序列鉴定  

Identification of Hudilong Origins by COⅠBarcoding and Characteristic Sequence

作　　者：李雷 史建伍[3] 李奇璋 吴宇 李耀昕[1] 张汉明 张磊[1] LI Lei;SHI Jian-wu;LI Qi-zhang;WU Yu;LI Yao-xin;ZHANG Han-ming;ZHANG Lei(School of Pharmacy,Naval Medical University(Second Military Medical University),Shanghai 200433,China;Unit 65655 of the People's Liberation Army of China,Chifeng 024000,China;Institute of Interdisciplinary Integrative Medicine Research,Medical School,Nantong University,Nantong 226001,China;Innovative Drug R&D Center,College of Life Sciences,Huaibei Normal University,Huaibei 235000,China)

机构地区：[1]海军军医大学(第二军医大学)药学院,上海200433 [2]中国人民解放军65655部队,内蒙古赤峰024000 [3]南通大学医学院交叉医学研究所,江苏南通226001 [4]淮北师范大学生命科学学院创新药物研发中心,安徽淮北235000

出　　处：《中国现代中药》2024年第10期1654-1662,共9页Modern Chinese Medicine

基　　金：国家自然科学基金项目(82225047);国家重点研发计划项目(2022YFC3501703)。

摘　　要：目的:补充线粒体细胞色素氧化酶Ⅰ(COⅠ)条形码鉴定沪地龙的种内、种间遗传信息并提取特征序列用于辅助鉴别,提高COⅠ条形码鉴定沪地龙基原的准确度。方法:收集不同产地沪地龙和其他品种蚯蚓,采用COⅠ条形码方法鉴定并获得鉴定序列,使用MEGA 7.0软件对所得序列进行系统发育树分析,计算各品种的种内、种间遗传距离,提取沪地龙3个基原的特征序列。结果:通俗环毛蚓与威廉环毛蚓的遗传距离为0.074,两者与栉盲环毛蚓间的遗传距离分别为0.187、0.194。经筛选整理后,共获得22个蚯蚓种类的271条序列,其中测序合并后得到序列218条、GenBank下载参考序列53条。发育树结果表明,不同蚯蚓种类在科与种一级均单独聚为一类,而属一级中,巨蚓科(Megascolecidae)腔蚓属(Metaphire)与远盲蚓属(Amynthas)未能较好区分。在通俗环毛蚓中提取了2条特征序列,威廉环毛蚓和栉盲环毛蚓中各提取了1条特征序列。结论:COⅠ条形码能够鉴定沪地龙与其他蚯蚓,特征序列能辅助提高沪地龙基原鉴定的效率和准确度。Objective:To supplement the genetic information for intraspecific and interspecific identification of Hudilong using COⅠbarcoding,and extract characteristic sequences to assist in identification,thereby improving the accuracy of COⅠbarcoding in identifying the origins of Hudilong.Methods:Samples of Hudilong from different regions and other varieties of earthworms were collected.Identification sequences were obtained using COⅠbarcoding,and phylogenetic tree analysis was conducted using MEGA 7.0 software.The intraspecific and interspecific genetic distances of each variety were calculated,and characteristic sequences were extracted from the three origins of Hudilong.Results:The genetic distance between Pheretima vulgaris and P.guillelmi was 0.074,while the genetic distances between these two species and P.pectinifera were 0.187 and 0.194,respectively.After screening and organizing the data,a total of 271 sequences from 22 species of earthworms were obtained,including 218 sequences from newly sequenced samples and 53 reference sequences downloaded from GenBank.The phylogenetic tree results showed that different earthworm species clustered separately at the family and species levels,but at the genus level,the genera Metaphire and Amynthas within the family Megascolecidae were not well differentiated.Two characteristic sequences were extracted from P.vulgaris,and one characteristic sequence each was extracted from P.guillelmi and P.pectinifera.Conclusions:COⅠbarcoding can differentiate Hudilong from other earthworms,and characteristic sequences can help improve the efficiency and accuracy of identifying the origins of Hudilong.

关 键 词：沪地龙 DNA条形码 特征序列 分子鉴定 

分 类 号：R282.5[医药卫生—中药学]