基于rbcL序列的大麻鉴定  

Identification of Cannabis Sativa L.Based on rbcL Sequence

作　　者：夏若成 张晓春 王笑笑 杨琪 陈冲 于欢 屈轶龄 王紫薇 施妍 向平 张素华 李成涛 XIA Ruo-cheng;ZHANG Xiao-chun;WANG Xiao-xiao;YANG Qi;CHEN Chong;YU Huan;QU Yi-ling;WANG Zi-wei;SHI Yan;XIANG Ping;ZHANG Su-hua;LI Cheng-tao(Department of Forensic Medicine,School of Basic Medical Sciences,Wenzhou Medical University,Wenzhou 325235,Zhejiang Province,China;Shanghai Key Laboratory of Forensic Medicine,Key Laboratory of Fo-rensic Science,Ministry of Justice,Shanghai Forensic Service Platform,Academy of Forensic Science,Shanghai 200063,China;Department of Forensic Medicine,Medical College of Soochow University,Suzhou 215123,Jiangsu Province,China;School of Forensic Medicine,Shanxi Medical University,Taiyuan 030001,China;School of Forensic Medicine,Xi’an Jiaotong University Health Science Center,Xi’an 710061,China)

机构地区：[1]温州医科大学基础医学院法医学系,浙江温州325235 [2]司法鉴定科学研究院,上海市法医学重点实验室司,法部司法鉴定重点实验室,上海市司法鉴定专业技术服务平台,上海200063 [3]苏州大学医学部法医学系,江苏苏州215123 [4]山西医科大学法医学院,山西太原030001 [5]西安交通大学医学部法医学院,陕西西安710061

出　　处：《法医学杂志》2021年第2期187-191,共5页Journal of Forensic Medicine

基　　金：上海市科委社会发展领域计划资助项目(19DZ1200600);国家自然科学基金资助项目(81930056,81625013);中央级科研院所公益项目(GY2020D-2);上海市法医学重点实验室资助项目(17DZ2273200);上海市司法鉴定专业技术服务平台资助项目(19DZ2292700);司法部司法鉴定重点实验室资助项目。

摘　　要：目的评估叶绿体DNA rbcL序列作为遗传标记鉴定大麻的可行性。方法检测62份大麻、10份啤酒花及10份葎草DNA的rbcL序列,并从GenBank数据库中下载96条大麻科rbcL序列。应用MEGA X软件进行序列比对,计算种内及种间Kimura-2-Parameter(K2P)遗传距离并构建系统聚类树。结果本次大麻及葎草属样本测序所得rbcL序列长度分别为617 bp和649 bp,且在所测大麻样本中检测到2种单倍型。BLAST相似性检索结果显示,测序所得序列与GenBank数据库中大麻rbcL序列相似性最高为100%。遗传距离分析结果显示,大麻种内不同样本间的最大遗传距离(0.0049)小于大麻与大麻科其他物种间的最小遗传距离(0.0129)。从中介网络图和系统聚类分析中可以看出,大麻与大麻科其他物种位于不同分支。结论rbcL序列可以作为鉴定大麻的DNA条形码,联合rbcL序列的比对分析和系统聚类分析有望成为法医学大麻种属鉴定可靠、便捷的检测手段。Objective To assess the feasibility of the rbcL sequence of chloroplast DNA as a genetic marker to identify Cannabis sativa L.Methods The rbcL sequences in 62 Cannabis sativa L.samples,10 Humulus lupulus samples and 10 Humulus scandens DNA samples were detected,and 96 rbcL se-quences of the Cannabaceae family were downloaded from Genbank.Sequence alignment was performed by MEGA X software,the intraspecific and interspecific Kimura-2-Parameter(K2P)genetic distances were calculated,and the system clustering tree was constructed.Results The rbcL sequence length ac-quired by sequencing of Cannabis sativa L.and Humulus scandens were 617 bp and 649 bp,respectively,and two haplotypes of Cannabis sativa L.were observed in the samples.The BLAST similarity search results showed that the highest similarity between the sequences acquired by sequencing and Cannabis sativa L.rbcL sequences available from Genbank was 100%.The genetic distance analysis showed that the maximum intraspecific genetic distance(0.0049)of Cannabis sativa L.was less than the minimum interspecific genetic distance(0.0129).The results of median-joining network and system clustering tree analysis showed that Cannabis sativa L.and other members of the Cannabaceae family were located in different branches.Conclusion The rbcL sequence could be used as a DNA barcode for identifying Cannabis sativa L.,and combined with comparative analysis of the rbcL sequence and system cluster analysis could be a reliable and effective detection method for Cannabis sativa L.identification in fo-rensic investigation.

关 键 词：法医遗传学 DNA测序 大麻 DNA条形码 RBCL序列 

分 类 号：DF795.2[医药卫生—法医学]