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 sequences 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 acquired 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 （0.004 9） of Cannabis sativa L. was less than the minimum interspecific genetic distance （0.012 9）. 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 forensic investigation.
题目: 基于rbcL序列的大麻鉴定.
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（0.004 9）小于大麻与大麻科其他物种间的最小遗传距离（0.012 9）。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。.
关键词: 法医遗传学；DNA测序；大麻；DNA条形码；rbcL序列.
Keywords: forensic genetics; DNA sequencing; Cannabis sativa L.; DNA barcode; rbcL sequence.
Copyright© by the Editorial Department of Journal of Forensic Medicine.