title: Use of Embryos Extracted from Individual Cannabis sativa Seeds for Genetic Studies and Forensic Applications.
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.affiliation: Institut de Conservació i Millora de l'Agrodiversitat Valenciana, Universitat Politècnica de València, Camí de Vera 14, València, 46022, Spain.
authors.forename: Salvador
authors.initials: S
authors.lastname: Soler
authors.affiliation: Institut de Conservació i Millora de l'Agrodiversitat Valenciana, Universitat Politècnica de València, Camí de Vera 14, València, 46022, Spain.
authors.forename: Dionís
authors.initials: D
authors.lastname: Borràs
authors.affiliation: Institut de Conservació i Millora de l'Agrodiversitat Valenciana, Universitat Politècnica de València, Camí de Vera 14, València, 46022, Spain.
authors.forename: Santiago
authors.initials: S
authors.lastname: Vilanova
authors.affiliation: Institut de Conservació i Millora de l'Agrodiversitat Valenciana, Universitat Politècnica de València, Camí de Vera 14, València, 46022, Spain.
authors.forename: Alicia
authors.initials: A
authors.lastname: Sifres
authors.affiliation: Institut de Conservació i Millora de l'Agrodiversitat Valenciana, Universitat Politècnica de València, Camí de Vera 14, València, 46022, Spain.
authors.forename: Isabel
authors.initials: I
authors.lastname: Andújar
authors.affiliation: Institut de Conservació i Millora de l'Agrodiversitat Valenciana, Universitat Politècnica de València, Camí de Vera 14, València, 46022, Spain.
authors.forename: Maria R
authors.initials: MR
authors.lastname: Figàs
authors.affiliation: Hemp Trading, S.L.U., Camí del Polio 51, Beniparrell, 46469, Spain.
authors.forename: Ernesto R
authors.initials: ER
authors.lastname: Llosa
authors.affiliation: Institut de Conservació i Millora de l'Agrodiversitat Valenciana, Universitat Politècnica de València, Camí de Vera 14, València, 46022, Spain.
authors.forename: Jaime
authors.initials: J
authors.lastname: Prohens
mesh_terms: D002188:Cannabis / Q000235:genetics*; D016172:DNA Fingerprinting / Q000379:methods; D018744:DNA, Plant / Q000302:isolation & purification*; D005787:Gene Frequency; D018895:Microsatellite Repeats; D016133:Polymerase Chain Reaction; D012639:Seeds / Q000235:genetics*
chemical_list: D018744:DNA, Plant
keywords: Cannabis sativa; DNA extraction; forensic science; marijuana; seed embryo; simple sequence repeats