Abstract
Genetic markers can be used in seeds and in plants to distinguish drug-type from fiber-type Cannabis Sativa L. varieties even at early stages, including pre-germination when cannabinoids are not accumulated yet. With this aim, this paper reports sequencing results for tetrahydrocannabinolic acid synthase (THCAS) and cannabidiolic acid synthase (CBDAS) genes from 21 C. sativa L. varieties. Taking into account that THCAS- and CBDAS-derived enzymes compete for the same substrate, the novelty of this work relies in the identification of markers based on both THCAS and CBDAS rather than THCAS alone. Notably, in our panel, we achieved an adequate degree of discrimination (AUC 100%) between drug-type and fiber-type cannabis samples. Our sequencing approach allowed identifying multiple genetic markers (single-nucleotide polymorphisms—SNPs—and a deletion/insertion) that effectively discriminate between the two subgroups of cannabis, namely fiber type vs. drug type. We identified four functional SNPs that are likely to induce decreased THCAS activity in the fiber-type cannabis plants. We also report the finding on a deletion in the CBDAS gene sequence that produces a truncated protein, possibly resulting in loss of function of the enzyme in the drug-type varieties. Chemical analyses for the actual concentration of cannabinoids confirmed the identification of drug-type rather than fiber-type genotypes. Genetic markers permit an early identification process for forensic applications while simplifying the procedures related to detection of therapeutic or industrial hemp.
Highlights
IntroductionCannabis sativa L. (commonly called cannabis) is an herbaceous plant belonging to the Cannabis genus of the Cannabaceae family
Cannabis sativa L. is an herbaceous plant belonging to the Cannabis genus of the Cannabaceae family
Genetic analyses analyses only only were were performed on a total of seeds: drug-type and fiber-type from the same varieties (Table performed on a total of 70 seeds: 20 drug-type and 50 fiber-type from the same varieties (Table S1)
Summary
Cannabis sativa L. (commonly called cannabis) is an herbaceous plant belonging to the Cannabis genus of the Cannabaceae family. (commonly called cannabis) is an herbaceous plant belonging to the Cannabis genus of the Cannabaceae family. The Cannabis genus includes morphologically variable varieties that can be mainly divided in two categories: Drug-type, suitable for recreational/therapeutic purposes, and fiber-type used in industrial and agronomy field. The different purposes of the cannabis genotypes depend on the ability of each variety or accession to synthesize and accumulate secondary metabolites known as cannabinoids. Cannabinoids represent a group of more than 100 natural products [1,2], of Plants 2019, 8, 496; doi:10.3390/plants8110496 www.mdpi.com/journal/plants. Plants 2019, 8, 496 which tetrahydrocannabinol (THC) is the main (psycho) active compound. Tetrahydrocannabinolic acid (THCA) and cannabidiolic acid (CBDA) are formed from the same precursor, namely cannabigerolic acid (CBGA). The THCA synthase (THCAS) and CBDA synthase (CBDAS) are necessary to produce the metabolites THCA and CBDA [3,4,5,6,7,8]
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