Evaluation of the trnK-matK-trnK, ycf3, and accD-psal chloroplast regions to differentiate crop type and biogeographical origin of Cannabis sativa
Evaluation of the trnK-matK-trnK, ycf3, and accD-psal chloroplast regions to differentiate crop type and biogeographical origin of Cannabis sativa
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DOI：
10.1007/s00414-021-02518-x
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发表时间：
2021-02-18
影响因子：
2.1
通讯作者：
Houston, Rachel
中科院分区：
文献类型：
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作者：
Cheng, Ya-Chih;Houston, Rachel
Cannabis sativa（大麻和大麻）是全球最具争议的作物之一。在美国，大麻的州特定合法化和最近合法化的大麻给执法带来了问题。本研究旨在利用叶绿体hSTR、INDEL和SNP标记来开发基因分型方法，以帮助区分法律的大麻与非法大麻，并追踪贩运大麻的流向。结果表明，该菌株在3个多态性区域：trnK-matK-trnK、ycf 3和accD-psal。为了区分作物类型和植物地理起源，对10种不同类型的水稻叶绿体基因组进行了分析。用定制片段和SNaPshot（TM）测定法对来自五个不同群体（来自美国和加拿大的大麻、来自智利和美国-墨西哥的大麻以及来自智利的医用大麻）的总共九个多态性位点进行基因分型。该研究还结合了来自同一样本集的基因型结果，使用了来自先前研究的21个额外的多态性标记。使用单倍型分析、系统发育分析、成对比较和主成分分析来评估这些多位点测定区分样品组的有效性。结果表明，一个明确的分离加拿大大麻只使用本研究中开发的9个多态性位点。当评估平均固定指数（F-ST）时，额外的21个标记物能够将美国大麻与两个大麻组分开至显著水平（p < 0.05）。本研究表明，这些细胞器标记可用于确定作物类型和C. sativa.然而，需要一个更广泛的数据库来评估这些标记的真正区分能力。
Cannabis sativa (marijuana and hemp) is one of the most controversial crops worldwide. In the USA, the state-specific legalization of marijuana and recently legalized hemp pose a problem for law enforcement. This study seeks to utilize chloroplast hSTRs, INDEL, and SNPs markers to develop genotyping methods to aid in the differentiation of legal hemp from illicit marijuana and also for tracking the flow of trafficked marijuana. Three polymorphic regions: trnK-matK-trnK, ycf3, and accD-psal, of the C. sativa chloroplast genome were evaluated in order to distinguish crop type and biogeographic origin. A total of nine polymorphic sites were genotyped from five distinct populations (hemp from the USA and Canada, marijuana from Chile and USA-Mexico, and medical marijuana from Chile) with a custom fragment and SNaPshot(TM) assay. The study also combined genotype results from the same sample set using 21 additional polymorphic markers from previous studies. The effectiveness of these multi-locus assays to distinguish sample groups was assessed using haplotype analysis, phylogenetic analysis, pairwise comparisons, and principal component analysis. Results indicated a clear separation of Canadian hemp using only the nine polymorphic sites developed in this study. The additional 21 markers were able to separate US hemp from both marijuana groups to a significant level (p < 0.05) when assessing average Fixation Indices (F-ST). This study demonstrated the applicability of these organelle markers for the determination of crop type and biogeographic origin of C. sativa. However, a more extensive database is needed to evaluate the true discriminatory power of these markers.