Field Testing of Collection Cards for Cannabis sativa Samples With a Single Hexanucleotide DNA Marker*,†

Allgeier, Lindsay; Hemenway, B S John; Shirley, B S Nicholas; LaNier, Tommy; Coyle, Heather Miller

doi:10.1111/j.1556-4029.2011.01818.x

Cited by 14 publications

(9 citation statements)

References 32 publications

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“…In 22 samples, the common two allele heterozygosity was not found; instead, three alleles were detected in at least one STR locus. Similar results were already found by previous studies [8,11,14]. The size of the alleles found was rounded…”

Section: Alleles and Genotypessupporting

confidence: 93%

The validation of a 15 STR multiplex PCR for Cannabis species

et al. 2012

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Trade and acquisition of Cannabis drugs are illegal in many countries worldwide; nevertheless, crimes related with these drugs are a major problem for the investigative authorities. With this manuscript, we want to introduce a 15 short tandem repeat (STR) Cannabis marker set that can be amplified in one PCR reaction. This multiplex PCR is specific to Cannabis species and combines highly informative STR markers. The 15 STR multiplex is easy to use and was validated according to common laboratory quality standards. Due to the fact that a lot of Cannabis plants are cultivated by clonal propagation and may show aneuploidy, polyploidy or multiple gene loci, it is not possible to apply biostatistics that follow the Hardy-Weinberg law. However, this multiplex will help the police to trace back trade routes of drug syndicates or dealers and it can help to link Cannabis plants to a crime scene.

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Section: Alleles and Genotypessupporting

confidence: 93%

The validation of a 15 STR multiplex PCR for Cannabis species

et al. 2012

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“…Among the genetic markers used to evaluate genetic diversity in Cannabis, SSRs (also called short tandem repeats, STS) have proved to be highly polymorphic (Gilmore et al, 2003;Howard et al, 2008;Allgeier et al, 2011;Shirley et al, 2013;Gao et al, 2014;Soler et al, 2016). Cannabis sativa genomic SSRs, which in general are more polymorphic than expressed sequence tag SSRs (EST-SSRs) (Eujayl et al, 2011;Muñoz-Falcón et al, 2011), can be obtained from previously published reports (Gilmore et al, 2003;Soler et al, 2016) or developed de novo from the available genome sequence of C. sativa (van Bakel et al, 2011).…”

Section: Introductionmentioning

confidence: 99%

Genetic structure of Cannabis sativa var. indica cultivars based on genomic SSR (gSSR) markers: Implications for breeding and germplasm management

Soler

Gramazio

Figàs

et al. 2017

Industrial Crops and Products

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Cannabis sativa L. is cultivated for its fiber or seeds (var. sativa; hemp), or for its high content in cannabinoids (var. indica; marijuana). Knowledge of the genetic structure of C. sativa var. indica is important for selection and breeding of cultivars with medicinal interest. We used six genomic SSRs (gSSRs) for genotyping 154 individual plants of 20 cultivars of C. sativa var. indica, plus two cultivars of C. sativa var. sativa. A very high polymorphism was observed, with an average of 17 alleles and 23.8 genotypes per locus. Expected (He) and observed (Ho) heterozygosities were high, with average values of 0.753 and 0.429, respectively. In some cultivars He and Ho presented similar values, while in others He was considerably higher than Ho suggesting that consanguinity and fixation had taken place during its development. In addition, some cultivars had a reduced number of alleles per locus (in some cases only two) indicating that a genetic bottleneck had taken place during its development. Gene flow (Nm) between both botanical varieties was high, with Nm=1.736. The molecular analysis of variance (AMOVA) revealed that only 31.94% of the molecular variation observed was caused by differences among cultivars, while the variation among plants of the same cultivar was of 37.11%, and within individual variation, due to heterozygosity, was of 30.96%. This indicates that a large variation exists within cultivars, which can be exploited for selection, but also complicates germplasm management and regeneration. The population structure analysis identified 14 genetic clusters, with most individuals of a single cultivar clustering together. This analysis, together with UPGMA cluster analysis shows that the two C. sativa var. sativa cultivars studied are differentiated from C. sativa var. indica, and that some cultivars of C. sativa var. indica seem to represent different selections from a common original cultivar. Our results represent the first comprehensive study of intra-3 varietal diversity in C. sativa var. indica and provide information of relevance for selection, breeding, and germplasm conservation, as well as for forensic studies in this crop.

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“…Many types of crime involve nonhuman DNA samples, such as those of animals, plants, pollen, bacterial and viruses [1-4]. Despite being a relatively new subdiscipline of forensic science, the interest of the forensic community regarding nonhuman DNA samples is considerable, as evidenced by [5].…”

Section: Introductionmentioning

confidence: 99%

Pet fur or fake fur? A forensic approach

Pilli

Casamassima²,

Vai

et al. 2014

Invest Genet

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BackgroundIn forensic science there are many types of crime that involve animals. Therefore, the identification of the species has become an essential investigative tool. The exhibits obtained from such offences are very often a challenge for forensic experts. Indeed, most biological materials are traces, hair or tanned fur. With hair samples, a common forensic approach should proceed from morphological and structural microscopic examination to DNA analysis. However, the microscopy of hair requires a lot of experience and a suitable comparative database to be able to recognize with a high degree of accuracy that a sample comes from a particular species and then to determine whether it is a protected one. DNA analysis offers the best opportunity to answer the question, ‘What species is this?’ In our work, we analyzed different samples of fur coming from China used to make hats and collars. Initially, the samples were examined under a microscope, then the mitochondrial DNA was tested for species identification. For this purpose, the genetic markers used were the 12S and 16S ribosomal RNA, while the hypervariable segment I of the control region was analyzed afterwards, to determine whether samples belonged to the same individual.ResultsMicroscopic examination showed that the fibres were of animal origin, although it was difficult to determine with a high degree of confidence which species they belonged to and if they came from a protected species. Therefore, DNA analysis was essential to try to clarify the species of these fur samples.ConclusionsMacroscopic and microscopic analysis confirmed the hypothesis regarding the analyzed hair belonging to real animals, although it failed to prove with any kind of certainty which actual family it came from, therefore, the species remains unknown. Sequence data analysis and comparisons with the samples available in GenBank showed that the hair, in most cases, belonged to the Canidae family, and in one case only to Felidae.

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Field Testing of Collection Cards for Cannabis sativa Samples With a Single Hexanucleotide DNA Marker*,†

Cited by 14 publications

References 32 publications

The validation of a 15 STR multiplex PCR for Cannabis species

The validation of a 15 STR multiplex PCR for Cannabis species

Genetic structure of Cannabis sativa var. indica cultivars based on genomic SSR (gSSR) markers: Implications for breeding and germplasm management

Pet fur or fake fur? A forensic approach

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