Forensic DNA Division, National Forensic Service, Wonju, 26460, Korea.
Forensic Toxicology Division, National Forensic Service, Wonju, 26460, Korea.
J Forensic Sci. 2020 Nov;65(6):2117-2120. doi: 10.1111/1556-4029.14515. Epub 2020 Jul 29.
Lophophora is a member of the Cactaceae family, which contains two species: Lophophora williamsii and L. diffusa. Lophophora williamsii is an illegal plant containing mescaline, a hallucinogenic alkaloid. In this study, a novel method based on a single nucleotide polymorphism (SNP) assay was developed for identifying L. williamsii; this assay reliably detects SNPs within chloroplast DNA (rbcL, matK, and trnL-trnF IGS) and was validated for identifying Lophophora and L. williamsii simultaneously. The chloroplast DNA sequences from four L. williamsii and three L. diffusa plants were obtained and compared using DNA sequence data from approximately 300 other Cactaceae species available in GenBank. From this sequence data, a total of seven SNPs were determined to be suitable for identifying L. williamsii. A multiplex assay was constructed using the ABI PRISM SNaPshot™ Multiplex Kit (Applied Biosystems, Forster City, CA) to analyze species-specific SNPs. Using this multiplex assay, we clearly distinguished the Lophophora among 19 species in the Cactaceae family. Additionally, L. williamsii was distinguished from L. diffusa. These results suggest that the newly developed assay may help resolve crimes related to illegal distribution and use. This multiplex assay will be useful for the genetic identification of L. williamsii and can complement conventional methods of detecting mescaline.
Lophophora williamsii 和 L. diffusa。Lophophora williamsii 是一种含有致幻生物碱麦斯卡林的非法植物。在这项研究中，开发了一种基于单核苷酸多态性 (SNP) 分析的鉴定 L. williamsii 的新方法；该方法可靠地检测叶绿体 DNA（rbcL、matK 和 trnL-trnF IGS）中的 SNPs，并已验证可同时鉴定龙舌兰属和 L. williamsii。从四个 L. williamsii 和三个 L. diffusa 植物中获得了叶绿体 DNA 序列，并与 GenBank 中大约 300 种其他仙人掌科物种的 DNA 序列数据进行了比较。从这些序列数据中，确定了总共 7 个 SNPs 可用于鉴定 L. williamsii。使用 ABI PRISM SNaPshot™ Multiplex Kit（Applied Biosystems，Forster City，CA）构建了多重分析，以分析物种特异性 SNPs。使用该多重分析，我们清楚地区分了仙人掌科 19 个物种中的龙舌兰属。此外，还可以区分 L. williamsii 和 L. diffusa。这些结果表明，新开发的分析方法可能有助于解决与非法分销和使用相关的犯罪。该多重分析将有助于鉴定 L. williamsii 的遗传特征，并可补充检测麦斯卡林的常规方法。
