Cannabis sativa, known by the common name marijuana, is the psychoactive drug most widely distributed in the world. Identification of Cannabis cultivars may be useful for association to illegal crops, which may reveal trafficking routes and related criminal groups. This study provides evidence for the performance of a segment of the rbcL gene, through genetic signature, as a tool for identification for C. sativa samples apprehended by the Rio de Janeiro Police, Brazil. The PCR amplified and further sequenced the fragment of approximately 561 bp of 24 samples of C. sativa rbcL gene and showed the same nucleotide sequences, suggesting a possible genetic similarity or identical varieties. Comparing with other Cannabaceae family sequences, we have found 99% of similarity between the Rio de Janeiro sequence and three other C. sativa rbcL genes. These findings suggest that the fragment utilized at this study is efficient in identifying C. sativa samples, therefore, useful in genetic discrimination of samples seized in forensic cases.
<em>Cannabis sativa</em> é uma das espécies mais antigas de plantas domesticadas e permanece como uma das culturas mais amplamente difundida e a droga ilícita mais consumida no mundo. Há uma grande difi culdade em identifi car e individualizar as amostras de Cannabis sp, difi cultando a correlação a prováveis locais de plantações ilegais, o que permitiria revelar rotas de tráfi co, relacionar grupos criminosos e distinguir amostras legais daquelas comercializadas como droga, onde o cultivo é permitido. A principal fi nalidade do DNA Barcode é o de proporcionar uma rápida e precisa identifi cação de organismos a partir de uma pequena região padronizada do genoma que ajuda a caracterizar e distinguir espécies e indivíduos não identifi cados para atribuir à espécie. Um dos genes candidatos é o ribulose-1,5-bisfosfato carboxilase/oxigenase (rbcL), utilizado como um sistema de DNA Barcode e presente no DNA dos cloroplastos das plantas, sendo o responsável pela produção subunidade maior da que converte dióxido de carbono e água em carboidratos.Nosso objetivo é o desenvolvimento de um protocolo efi ciente de extração de DNA e de sequenciamento do gene rbcL para análise forense de amostras apreendidas da Polícia Judiciária. DNA de amostras de Cannabis sativa,caracterizadas no ICCE/DGPTC/PCERJ, foi extraído utilizandoo Mini DNeasy Plant (Qiagen)no IPPGF/DGPTC/PCERJ. O material foi transferido para o LabFor/UFRJe um fragmento de 735 pb do gene rbcL foi amplifi cado através de um par de iniciadores descritos na literatura como universais para plantas.O fragmento foi sequenciado, com o protocolo do BigDye v3.1, usando ABI 3500 (Life Technologies). As comparações das sequências foram realizadas no software Geneious (Biomatters). A nossa sequência consenso de 559 pb foi comparada às seqüências que correspondem às regiões rbcL em <em>Cannabis sativa</em> depositadas no GenBank, e observamos a ocorrência de polimorfi smos (SNPs) entre as amostras dos Estados unidos, Reino Unido e China, sugerindo que se trata de uma assinatura genética para análise forense.
The use of bi-allelic markers such as retrotransposable element insertion polymorphisms or Innuls (for insertion/null) can overcome some limitations of short tandem repeat (STR) loci in typing forensic biological evidence. This study investigated the efficiency of the InnoTyper® 21 Innul markers in an urban admixed population sample in Rio de Janeiro (n = 40) and one highly compromised sample collected as evidence by the Rio de Janeiro police. No significant departures from Hardy-Weinberg equilibrium were detected after the Bonferroni correction (α' ≈ 0.05/20, p < 0.0025), and no significant linkage disequilibrium was observed between markers. Assuming loci independence, the cumulative random match probability (RMP) was 2.3 × 10. A lower mean Fis value was obtained for this sample population compared with those of three North American populations (African-American, Southwest Hispanic, US Caucasian). Principal component analysis with the three North American populations and one from 21 East Asian population showed that African Americans segregated as an independent group while US Caucasian, Southwest Hispanic, East Asian, and Rio de Janeiro populations are in a single large heterogeneous group. Also, a full Innuls profile was produced from an evidence sample, despite the DNA being highly degraded. In conclusion, this system is a useful complement to standard STR kits.
The majority of STR loci are not ideal for the analysis of forensic samples with degraded and/or low template DNA. One alternative to overcome these limitations is the use of bi-allelic markers, which have low mutation rates and shorter amplicons. Human identification (HID) InDel marker panels have been described in several countries, including Brazil. The commercial kit available is, however, mostly suitable for Europeans, with lower discrimination power for other population groups. Recently, a combination of 49 InDel markers used in four different ethnic groups in the USA has been shown to be more informative than another panel from Portugal, already tested in a Rio de Janeiro sample. However, these 49 InDels have yet to be applied to other admixed or isolated populations. We assessed the efficiency of this panel in two urban admixed populations (Rio de Janeiro, Brazil; Tripoli, Libya) and one isolated Native Brazilian community. All markers are in Hardy-Weinberg equilibrium (HWE) after the Bonferroni correction, and no Linkage disequilibrium was detected. Assuming loci independence and no substructure effect, cumulative RMP was 2.7×10(-18), 1.5×10(-20), and 4.5×10(-20) for Native Brazilian, Rio de Janeiro, and Tripoli populations, respectively. The overall Fst value was 0.05512. Rio de Janeiro and Tripoli showed similar admixture levels, however for Native Brazilians one parental cluster represented over 60 % of the total parental population. We conclude that this panel is suitable for HID on these urban populations, but is less efficient for the isolated group.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.