Philippe Henry1,2, Surender Khatodia3, Karan Kapoor3, Britni Gonzales4, Alexis Middleton4, Kevin Hong4, Aaron Hilyard4, Steve Johnson4, Davis Allen5, Zachary Chester5, Dan Jin6,7,8, José Carlos Rodriguez Jule9, Iain Wilson9, Manu Gangola10, Jason Broome10,11, Deron Caplan10, Dinesh Adhikary12, Michael K Deyholos12, Michael Morgan12,13, Oliver W Hall13, Brent J Guppy14,15,16, Cindy Orser4. 1. VSSL Enterprises Ltd., West Kelowna, BC, Canada. philippe@egret.bio. 2. Digipath Labs Inc., Las Vegas, NV, USA. philippe@egret.bio. 3. VSSL Enterprises Ltd., West Kelowna, BC, Canada. 4. Digipath Labs Inc., Las Vegas, NV, USA. 5. Island Genetics Ltd., Vancouver, BC, Canada. 6. Biomedical Engineering, University of Alberta, Edmonton, AB, Canada. 7. Polar Bear Genome BioPharma, Edmonton, AB, Canada. 8. Labs-Mart Inc., Edmonton, AB, Canada. 9. Okanagan Gold Cannabis Corp, West Kelowna, BC, Canada. 10. The Flowr Group (Okanagan) Inc., Kelowna, BC, Canada. 11. HYTN Beverages, Vancouver, BC, Canada. 12. Biology, The University of British Columbia Okanagan, Kelowna, BC, Canada. 13. Noble Growth Corp, Drayton Valley, AB, Canada. 14. Synthase Genetics Inc., Winnipeg, MB, Canada. 15. OneLeaf Cannabis Co., Regina, Saskatchewan, Canada. 16. Botanist Organic Growers, Winnipeg, Manitoba, Canada.
Abstract
BACKGROUND: The taxonomic classification of Cannabis genus has been delineated through three main types: sativa (tall and less branched plant with long and narrow leaves), indica (short and highly branched plant with broader leaves) and ruderalis (heirloom type with short stature, less branching and small thick leaves). While still under discussion, particularly whether the genus is polytypic or monotypic, this broad classification reflects putative geographical origins of each group and putative chemotype and pharmacologic effect. METHODS: Here we describe a thorough investigation of cannabis accessions using a set of 23 highly informative and polymorphic SNP (Single Nucleotide Polymorphism) markers associated with important traits such as cannabinoid and terpenoid expression as well as fibre and resin production. The assay offers insight into cannabis population structure, phylogenetic relationship, population genetics and correlation to secondary metabolite concentrations. We demonstrate the utility of the assay for rapid, repeatable and cost-efficient genotyping of commercial and industrial cannabis accessions for use in product traceability, breeding programs, regulatory compliance and consumer education. RESULTS: We identified 5 clusters in the sample set, including industrial hemp (K5) and resin hemp, which likely underwent a bottleneck to stabilize cannabidiolic acid (CBDA) accumulation (K2, Type II & III). Tetrahydrocannabinolic acid (THCA) resin (Type I) makes up the other three clusters with terpinolene (K4 - colloquial "sativa" or "Narrow Leaflet Drug" (NLD), myrcene/pinene (K1) and myrcene/limonene/linalool (K3 - colloquial "indica", "Broad Leaflet Drug" (BLD), which also putatively harbour an active version of the cannabichrometic acid Synthase gene (CBCAS). CONCLUSION: The final chemical compositions of cannabis products have key traits related to their genetic identities. Our analyses in the context of the NCBI Cannabis sativa Annotation Release 100 allows for hypothesis testing with regards to secondary metabolite production. Genetic markers related to secondary metabolite production will be important in many sectors of the cannabis marketplace. For example, markers related to THC production will be important for adaptable and compliant large-scale seed production under the new US Domestic Hemp Production Program.
BACKGROUND: The taxonomic classification of Cannabis genus has been delineated through three main types: sativa (tall and less branched plant with long and narrow leaves), indica (short and highly branched plant with broader leaves) and ruderalis (heirloom type with short stature, less branching and small thick leaves). While still under discussion, particularly whether the genus is polytypic or monotypic, this broad classification reflects putative geographical origins of each group and putative chemotype and pharmacologic effect. METHODS: Here we describe a thorough investigation of cannabis accessions using a set of 23 highly informative and polymorphic SNP (Single Nucleotide Polymorphism) markers associated with important traits such as cannabinoid and terpenoid expression as well as fibre and resin production. The assay offers insight into cannabis population structure, phylogenetic relationship, population genetics and correlation to secondary metabolite concentrations. We demonstrate the utility of the assay for rapid, repeatable and cost-efficient genotyping of commercial and industrial cannabis accessions for use in product traceability, breeding programs, regulatory compliance and consumer education. RESULTS: We identified 5 clusters in the sample set, including industrial hemp (K5) and resin hemp, which likely underwent a bottleneck to stabilize cannabidiolic acid (CBDA) accumulation (K2, Type II & III). Tetrahydrocannabinolic acid (THCA) resin (Type I) makes up the other three clusters with terpinolene (K4 - colloquial "sativa" or "Narrow Leaflet Drug" (NLD), myrcene/pinene (K1) and myrcene/limonene/linalool (K3 - colloquial "indica", "Broad Leaflet Drug" (BLD), which also putatively harbour an active version of the cannabichrometic acid Synthase gene (CBCAS). CONCLUSION: The final chemical compositions of cannabis products have key traits related to their genetic identities. Our analyses in the context of the NCBI Cannabis sativa Annotation Release 100 allows for hypothesis testing with regards to secondary metabolite production. Genetic markers related to secondary metabolite production will be important in many sectors of the cannabis marketplace. For example, markers related to THC production will be important for adaptable and compliant large-scale seed production under the new US Domestic Hemp Production Program.
Entities:
Keywords:
Cannabinoids; Cannabis; Compliance; Genetic assay; Hemp; Population structure; Terpenes
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