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Use of Embryos Extracted from Individual Cannabis sativa Seeds for Genetic Studies and Forensic Applications.
J Forensic Sci. 2016 Mar; 61(2):494-500.JF

Abstract

Legal limits on the psychoactive tetrahydrocannabinol (THC) content in Cannabis sativa plants have complicated genetic and forensic studies in this species. However, Cannabis seeds present very low THC levels. We developed a method for embryo extraction from seeds and an improved protocol for DNA extraction and tested this method in four hemp and six marijuana varieties. This embryo extraction method enabled the recovery of diploid embryos from individual seeds. An improved DNA extraction protocol (CTAB3) was used to obtain DNA from individual embryos at a concentration and quality similar to DNA extracted from leaves. DNA extracted from embryos was used for SSR molecular characterization in individuals from the 10 varieties. A unique molecular profile for each individual was obtained, and a clear differentiation between hemp and marijuana varieties was observed. The combined embryo extraction-DNA extraction methodology and the new highly polymorphic SSR markers facilitate genetic and forensic studies in Cannabis.

Authors+Show Affiliations

Institut de Conservació i Millora de l'Agrodiversitat Valenciana, Universitat Politècnica de València, Camí de Vera 14, València, 46022, Spain.Institut de Conservació i Millora de l'Agrodiversitat Valenciana, Universitat Politècnica de València, Camí de Vera 14, València, 46022, Spain.Institut de Conservació i Millora de l'Agrodiversitat Valenciana, Universitat Politècnica de València, Camí de Vera 14, València, 46022, Spain.Institut de Conservació i Millora de l'Agrodiversitat Valenciana, Universitat Politècnica de València, Camí de Vera 14, València, 46022, Spain.Institut de Conservació i Millora de l'Agrodiversitat Valenciana, Universitat Politècnica de València, Camí de Vera 14, València, 46022, Spain.Institut de Conservació i Millora de l'Agrodiversitat Valenciana, Universitat Politècnica de València, Camí de Vera 14, València, 46022, Spain.Hemp Trading, S.L.U., Camí del Polio 51, Beniparrell, 46469, Spain.Institut de Conservació i Millora de l'Agrodiversitat Valenciana, Universitat Politècnica de València, Camí de Vera 14, València, 46022, Spain.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

27404624

Citation

Soler, Salvador, et al. "Use of Embryos Extracted From Individual Cannabis Sativa Seeds for Genetic Studies and Forensic Applications." Journal of Forensic Sciences, vol. 61, no. 2, 2016, pp. 494-500.
Soler S, Borràs D, Vilanova S, et al. Use of Embryos Extracted from Individual Cannabis sativa Seeds for Genetic Studies and Forensic Applications. J Forensic Sci. 2016;61(2):494-500.
Soler, S., Borràs, D., Vilanova, S., Sifres, A., Andújar, I., Figàs, M. R., Llosa, E. R., & Prohens, J. (2016). Use of Embryos Extracted from Individual Cannabis sativa Seeds for Genetic Studies and Forensic Applications. Journal of Forensic Sciences, 61(2), 494-500. https://doi.org/10.1111/1556-4029.12995
Soler S, et al. Use of Embryos Extracted From Individual Cannabis Sativa Seeds for Genetic Studies and Forensic Applications. J Forensic Sci. 2016;61(2):494-500. PubMed PMID: 27404624.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Use of Embryos Extracted from Individual Cannabis sativa Seeds for Genetic Studies and Forensic Applications. AU - Soler,Salvador, AU - Borràs,Dionís, AU - Vilanova,Santiago, AU - Sifres,Alicia, AU - Andújar,Isabel, AU - Figàs,Maria R, AU - Llosa,Ernesto R, AU - Prohens,Jaime, Y1 - 2015/12/22/ PY - 2014/12/22/received PY - 2015/04/19/revised PY - 2015/04/26/accepted PY - 2016/7/13/entrez PY - 2016/7/13/pubmed PY - 2017/1/19/medline KW - Cannabis sativa KW - DNA extraction KW - forensic science KW - marijuana KW - seed embryo KW - simple sequence repeats SP - 494 EP - 500 JF - Journal of forensic sciences JO - J Forensic Sci VL - 61 IS - 2 N2 - Legal limits on the psychoactive tetrahydrocannabinol (THC) content in Cannabis sativa plants have complicated genetic and forensic studies in this species. However, Cannabis seeds present very low THC levels. We developed a method for embryo extraction from seeds and an improved protocol for DNA extraction and tested this method in four hemp and six marijuana varieties. This embryo extraction method enabled the recovery of diploid embryos from individual seeds. An improved DNA extraction protocol (CTAB3) was used to obtain DNA from individual embryos at a concentration and quality similar to DNA extracted from leaves. DNA extracted from embryos was used for SSR molecular characterization in individuals from the 10 varieties. A unique molecular profile for each individual was obtained, and a clear differentiation between hemp and marijuana varieties was observed. The combined embryo extraction-DNA extraction methodology and the new highly polymorphic SSR markers facilitate genetic and forensic studies in Cannabis. SN - 1556-4029 UR - https://www.unboundmedicine.com/medline/citation/27404624/Use_of_Embryos_Extracted_from_Individual_Cannabis_sativa_Seeds_for_Genetic_Studies_and_Forensic_Applications_ L2 - https://doi.org/10.1111/1556-4029.12995 DB - PRIME DP - Unbound Medicine ER -