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Cannabis is an ancient crop representing a rapidly increasing legal market, especially for medicinal purposes. Medicinal and psychoactive effects of Cannabis rely on specific terpenophenolic ligands named cannabinoids. Recent whole-genome sequencing efforts have uncovered variation in multiple genes encoding the final steps in cannabinoid biosynthesis. However, the origin, evolution, and phylogenetic relationships of these cannabinoid oxidocyclase genes remain unclear. To elucidate these aspects, we performed comparative genomic analyses of Cannabis , related genera within the Cannabaceae family, and selected outgroup species. Results show that cannabinoid oxidocyclase genes originated in the Cannabis lineage from within a larger gene expansion in the Cannabaceae family. Localization and divergence of oxidocyclase genes in the Cannabis genome revealed two main syntenic blocks, each comprising tandemly repeated cannabinoid oxidocyclase genes. By comparing these blocks with those in genomes from closely related species, we propose an evolutionary model for the origin, neofunctionalization, duplication, and diversification of cannabinoid oxidocycloase genes. Based on phylogenetic analyses, we propose a comprehensive classification of three main clades and seven subclades that are intended to aid unequivocal referencing and identification of cannabinoid oxidocyclase genes. Significance statement. Cannabis genome sequencing efforts have revealed extensive cannabinoid oxidocyclase gene variation. However, phylogenetic relationships and evolution of these genes remain unclear. Our comprehensive analysis of currently available data reveals that these genes comprise three main clades and seven subclades that originated through Cannabis -specific gene duplication and divergence. Our new conceptual and evolutionary framework serves as a reference for future description and functional analyses of cannabinoid oxidocyclases. Many of the medicinal properties of Cannabis are due to its production of cannabinoids; a unique class of psychoactive terpenophenolic ligands Gaoni and Mechoulam ; Mechoulam It is important to note, however, that cannabinoids are synthesized and stored in the plant as acids that are not medicinally active. Only from exposure to light during storage or heat during processing for consumption e. Some other plant genera such as Rhododendron and Radula have also been found to make cannabinoids Iijima et al. THC is responsible for the psychoactive effect of Cannabis through its partial agonist activity at endocannabinoid receptors Gaoni and Mechoulam ; Mechoulam and Parker This effect is the reason for the large-scale use of Cannabis as an intoxicant. But accumulating evidence from clinical trials indicates that moderate doses of THC can be used medicinally to, for example, reduce nausea and vomiting, pain, and improvement of sleep and appetite van de Donk et al. CBD has a weak affinity for endocannabinoid receptors and is not psychoactive Pertwee It has been found to modulate the effects of THC and endocannabinoids and may be effective for symptomatic treatment of anxiety and psychosis and in treating some childhood epilepsy syndromes Gofshteyn et al. However, cultivars exist with alternative chemical profiles such as drug-type cultivars with high levels of CBD and other classifications based on chemotype have been proposed Hazekamp et al. These lesser-known cannabinoids may have anti-inflammatory effects but the evidence is relatively scarce Brierley et al. Within the Cannabis plant, cannabinoids are synthesized in multicellular epidermal glands glandular trichomes that are most abundant on the bracts of female inflorescences. The cannabinoid biosynthetic pathway has been largely elucidated, and for many steps in the pathway, the corresponding enzymes have been isolated and characterized fig. In brief, cannabinoid biosynthesis relies on two precursors from two distinct metabolic pathways: olivetolic acid from the polyketide pathway and geranyl-geranyl pyrophosphate GPP from the methylerythritol phosphate MEP pathway. CBGA is then secreted into the extracellular storage cavity via an unknown mechanism and further processed by secreted cannabinoid oxidocyclases that perform different types of oxidative cyclizations of its linear prenyl moiety into derived cannabinoid acids such as tetrahydrocannabinolic acid THCA , cannabidiolic acid CBDA , and cannabichromenic acid CBCA Taura et al. Cannabinoid biosynthesis pathway. Dotted arrows indicate nonenzymatic decarboxylations; solid arrows indicate enzymatic reactions; enzyme names are shown in blue, while resulting compounds are shown in black. Compound sub structures depicted in red signify those that represent a single unit of GPP. This family is named after an oxidocyclase from Eschscholzia californica involved in alkaloid biosynthesis and part of the larger oxygen-dependent FAD-linked oxidoreductase family PF Hauschild et al. However, the latest phylogenetic classification of plant BBE-like genes was based on Arabidopsis sequences only Brassicaceae Daniel et al. Even though some BBE-like enzymes related to cannabinoid oxidocyclases have been identified Aryal et al. Therefore, a comprehensive phylogenetic analysis of BBE-like enzymes including cannabinoid oxidocyclase genes is warranted. Although environmental factors play a role in determining the amount of cannabinoids present in different parts and stages of the plant Rustichelli et al. However, later genome sequencing revealed that they are encoded by different genes rather than alleles within a large polymorphic genomic region with low levels of recombination Kojoma et al. Thus, they are treated as separate genes below. For this reason, the gene has been used as a diagnostic marker for detecting psychoactive cultivars for crop breeding and forensics Kojoma et al. This is probably due to a single amino acid mutation leading to a defective B T0 variant Onofri et al. Thus, it remains unclear if THCAS occurs in multiple copies and, consequently, if copy number variation could be a target for the breeding of cultivars. However, different missense mutations have been described from CBGA-dominant i. Indeed, it could be used to discriminate between fiber and drug-type cultivars as well as accurately predict chemotype in feral and cultivated plants Cascini et al. Notably, up to three different variants of such putative pseudogenes were detected in single cultivars van Bakel et al. Other reported amplified fragments may also represent the same gene. These may encode enzymes for other cannabinoids, but their copy numbers and sequence properties are not well described or cataloged Hurgobin et al. Weiblen et al. But, it remains unclear how these relate to the gene fragments listed above and to each other. The total number of cannabinoid oxidocyclase genes varies considerably across cultivars. Onofri et al. McKernan et al. A recent study on copy number variation in cannabinoid oxidocyclase genes estimated that some of the analyzed cultivars could have up to 10 different fragments Vergara et al. Based on these results it is clear that cannabinoid oxidocyclase genes can be considered a unique gene family that stems from a recent expansion and includes genes with unknown function Onofri et al. However, due to differences in 1 primers used for amplification, 2 reference genomes used for copy number estimation, and 3 level of homozygosity, these numbers are not directly comparable and may not be accurate assessments of gene copy number. There is also no appropriate classification to facilitate the unequivocal naming and referencing of cannabinoid oxidocyclase genes. Finally, it remains unclear whether these genes are specific to Cannabis. A more recent phylogenetic analysis based on genomic data applied the same a priori assumption Hurgobin et al. Therefore, it remains unknown whether cannabinoid oxidocyclase genes are specific to Cannabis or represent more ancient gene duplications in, for example, an ancestor of Cannabis and related genera within the Cannabaceae family such as Humulus and Trema Padgitt-Cobb et al. In addition, the genomic localization of many described gene sequences remains unknown and, consequently, we lack a clear overview of the patterns of gene duplication and divergence across the Cannabis genome Weiblen et al. We present a comparative analysis of cannabinoid oxidocyclase genes in the genomes of Cannabis , closely related genera and informative outgroup species. This was greatly aided by the recent release and publication of several diverse Cannabis genome assemblies based on long-read sequencing technologies McKernan et al. Gao et al. In addition, genomic information is available for other genera in the Cannabaceae family. Recent species-level phylogenetic analyses of the Cannabaceae family based on plastome sequences suggest that the genera Parasponia and Trema together are sister to Cannabis and Humulus Jin et al. Draft genome assemblies have recently become available for Humulus , Parasponia , and Trema , that can be used for comparative analyses of Cannabis genes van Velzen et al. This provides an excellent opportunity to perform a comprehensive reconstruction of the evolution of cannabinoid oxidocyclase genes. In addition, Morus notabilis Moraceae , Medicago truncatula Fabaceae , and Arabidopsis thaliana Brassicaceae were included as outgroups, allowing us to place our results within a broader phylogenetic perspective and in the context of the existing BBE-like gene family classification Daniel et al. Based on phylogenetic and synteny analysis, we elucidate the evolution of these genes and address the following questions:. How are cannabinoid oxidocyclases related to other berberine bridge enzymes? Are cannabinoid oxidocyclase genes specific to Cannabis or do they represent more ancient duplications in, for example, an ancestor of Cannabis and related genera within the Cannabaceae family? What are the patterns of duplication and divergence of cannabinoid oxidocyclase genes across Cannabis genomes? We also present a comprehensive clade-based classification of all cannabinoid oxidocyclase genes to resolve current confusion due to inconsistencies in naming and aid their future referencing and identification. To place cannabinoid oxidocyclase genes within the context of the BBE-like gene family we performed a phylogenetic analysis of BBE-like protein sequences from selected Eurosid genomes supplementary table S1 , Supplementary Material online. These include genomes from C. Genomes from Morus notabilis Moraceae , Medicago truncatula Fabaceae , and Arabidopsis thaliana Brassicaceae were included as outgroups fig. The resulting gene tree recovered 11 clades, including groups 1—7 earlier described Daniel et al. Cannabis BBE-like sequences were found in groups 2, 5. Within this group, a Cannabaceae-specific gene expansion can be identified within which all three known cannabinoid oxidocyclase occur in a Cannabis -specific clade, which we name the cannabinoid oxidocyclase clade. Eurosid berberine bridge enzyme gene family analysis. A Gene tree based on protein sequences showing that cannabinoid oxidocyclases comprise a Cannabis -specific clade. Colored blocks indicate the identified groups 1—12; node labels indicate posterior probabilities below 1. Bottom right inset shows known relationships among sampled species. B genomic colocalization of berberine bridge enzymes in C. Grey horizontal bars indicate contigs in the Cannabis chromosomal scaffold shown in figure 4A. Vertical lines indicate locations of annotated genes; berberine bridge enzymes are indicated with triangles in color and numbering consistent with those in A. For displaying purposes, genomic scaffolds are not shown in the same scale size shown is indicated. We therefore retrieved genomic locations of all BBE-like genes in other genomes including T. This revealed that BBE-like genes from different clades are commonly colocalized in these genomes fig. This suggests that selection favors BBE-like genes to remain in close genomic proximity. It is known that genes involved in the same pathway have the tendency to cluster in plant genomes Liu et al. However, it is not clear if and how the various BBE-like genes share pathways and we therefore have no conclusive explanation for this intriguing pattern. Additional analyses based on sequences from NCBI and from more fragmented Cannabis genomes are shown in supplementary figures S1 and S2 , Supplementary Material online, respectively. Based on the resulting gene trees we consistently recovered the same three main clades A—C; fig. Cannabinoid oxidocyclase gene tree. Labels indicate genbank accession of genomic contig and locus tag when available or start position. Sequences included in the BBE-like analysis shown in figure 2A are marked with an asterisk. Colored blocks indicate the identified clades; white blocks indicate sequence types. Node labels indicate posterior probabilities below 1. It comprises four subclades and can be characterized by three unique nonsynonymous substitutions supplementary table S2 , Supplementary Material online. Group 2 comprises six types sharing the nonsynonymous substitution C Leu. Other types remain ungrouped. Type 3 Onofri et al. Type 4 Onofri et al. Type 5 comprises partial sequences from various regions in Pakistan described by Ali et al. They can be characterized by two unique aa substitutions: T Val and C Pro. Type 6 comprises accession MT Type 7 comprises accession LC Type 8 comprises the putative THCAS sequence of a putatively wild plant from Jilong, Tibet that can be characterized by six unique aa substitutions. They have not yet been functionally assessed but given that at least one variant comprises a full-length coding sequence it is expected to have some functional relevance; probably as a cannabinoid oxidocyclase. Clade B comprises two subclades and can be characterized by 16 unique aa substitutions supplementary table S2 , Supplementary Material online. S1 and S2 , Supplementary Material online; table 1. They can be further divided into two types that correspond with groups 5 and 6 described by Onofri et al. We note that some sequences described by Cascini et al. S1 and S2 , Supplementary Material online. The first type can be characterized by four secondary nonsense mutations and eight secondary unique aa substitutions supplementary table S2 , Supplementary Material online. The second type can be characterized by two secondary unique aa substitutions. The third type can be characterized by nine secondary unique aa substitutions supplementary table S2 , Supplementary Material online. Accession LKUA They share 19 unique aa substitutions and can be divided into seven types fig. Based on nucleotide alignments and protein comparisons, we found that all cannabinoid oxidocyclase genes occur in two main syntenic clusters, together with other BBE-like genes. In the chemotype II Jamaican Lion genome, there are two putative allelic variants; the first comprising two full-length coding sequences and two nonfunctional pseudo gene copies and the second comprising five full-length coding sequences and one nonfunctional pseudo gene copy. S3 , Supplementary Material online. All variants comprise another copy of a group 5. The first variant comprises a single copy of THCAS , a tandemly repeated array of subclade B2 nonfunctional pseudo genes, and a nonfunctional pseudo gene from the A4 subclade. This suggests high levels of divergence across this large genomic region. Cannabinoid oxidocyclase microsynteny assessments. Triangles indicate genes not to scale colored according to their homology and putative orthologs are connected with colored lines. Nonfunctional pseudo genes are shown without black outlines. Cannabinoid oxidocyclase genes are members of BBE-like group 10 see fig. S4 , Supplementary Material online. Some of these flanking genes are considered pathogen response genes McKernan et al. Interestingly, it also includes a nonfunctional pseudo gene from subclade A3 but given the lack of additional A3 copies within the array, this appears to be a relatively recent insertion. No further synteny was found with Humulus , Parasponia , or Trema ; suggesting that this syntenic cluster is specific to Cannabis. To assess the direction of evolution, we then assessed protein-level microsynteny in genomes from closely related Cannabaceae species H. This revealed that each comprises a tandemly repeated array of group 10 BBE-like genes that are closely related to the known cannabinoid oxidocyclase genes, as well as a single copy of the group 5. This suggests that cannabinoid oxidocyclases originated within an ancestral syntenic block and experienced a series of tandem gene duplications, translocations, and divergence. Since the cannabinoid oxidocyclase genes were first discovered and described, it has been known that they are members of the BBE-like gene family Sirikantaramas et al. However, the BBE-like family is large and the most recent classification of plant BBE-like genes was based only on analysis of genes from Arabidopsis in the Brassicaceae family Daniel et al. Our results show that cannabinoid oxidocyclase genes from Cannabis originated from a newly defined clade Group 10 within the BBE-like gene family fig. Within Group 10 gene expansions occurred independently in Moraceae and Cannabaceae fig. The expansion in Cannabaceae eventually led to the origin of cannabinoid oxidocyclases. Such gene diversification and enzymatic versatility confirm that BBE-like enzymes play important roles in generating biochemical novelty Daniel et al. These results show unequivocally and for the first time that cannabinoid oxidocyclase genes did not originate from more ancient duplications within the Cannabaceae but are specific to Cannabis. We therefore hypothesize that the CBGA biosynthetic pathway existed before the origin of cannabinoid oxidocyclases. The comparative approach that we leveraged here can help elucidate the order in which these pathway genes evolved and, thus, reconstruct the origin of a novel and societally relevant biosynthetic pathway. Based on our phylogenetic analysis, we classified the cannabinoid oxidocyclase genes into three main clades A—C comprising a total of seven sub clades fig. In addition to these three subclades representing functionally characterized cannabinoid oxidocyclase genes, we identified four previously unrecognized subclades. Based on current sampling, two of these clades contain only pseudogenes. Within subclade A4, two types can be recognized that share four nonsense mutations. Similarly, within subclade B2, three types can be recognized that share a frame-shift mutation supplementary table S2 , Supplementary Material online. Therefore, it seems that within each of these two subclades, the most recent common ancestor was likely already nonfunctional. Contrastingly, clade C and subclade A3 each include full-length coding sequences that are most likely functional enzymes. Taura et al. It has not yet been experimentally tested. Consequently, there is potential that the products of clade C and subclade A3-encoded enzymes are of biochemical and potential medical importance. Our clade-based classification is intended to aid unequivocal referencing and identification of cannabinoid oxidocyclase genes. In retrospect, much of the confusion about gene identity stems from the general tendency to name sequences in accordance with the primers used for their amplification. These coamplifications are undoubtedly due to the high levels of sequence similarity between these genes. We anticipate that our classification will help avoid such confusion about the identity and relationships of cannabinoid oxidocyclase genes in the future. Our comprehensive analyses sampling all currently available sequences consistently recovered the same clades see fig. S1 and S2 , Supplementary Material online , suggesting that this classification is robust. New cannabinoid oxidocyclase sequences can be associated with the corresponding clade by phylogenetic analysis or based on the clade-specific missense mutations listed in supplementary table S2 , Supplementary Material online. In case sequences fall outside any of our described clades, new sub clades can be defined in accordance with our system. However, the genomic locations of most other oxidocyclase genes have remained unknown. Consequently, a comprehensive overview of the patterns of gene duplication and divergence across the Cannabis genome has been lacking Weiblen et al. Our assessment of microsynteny based on nucleotide alignments and protein comparisons revealed that cannabinoid oxidocyclase genes occur in two large syntenic blocks. Similarly, it is yet unclear if CBDAS and the subclade B2 pseudogenes are orthologous or comprise different paralogous loci. Our protein-level microsynteny analysis including genomes from closely related species H. Below, we propose our most parsimonious evolutionary interpretation of cannabinoid oxidocyclase gene duplication and divergence fig. Subsequent gene duplication and divergence lead to a set of ancestral genes representing the three main extant clades A, B, and C. Next, tandem duplication of this set together with the closely linked group 5. Block 1 retained ancestral genes representing clades A diverging into an ancestor of the extant clade A4 pseudogenes and C; clade B was apparently lost. Block 2 retained ancestral genes representing clades A an ancestral clade A3 gene originated through duplication and divergence and B; clade C was apparently lost. Support for this hypothetical tandem duplication event can be found in the BBE-like gene tree where the two corresponding group 5. Finally, large-scale divergence of the second block led to the three variants of the divergent region described above. Evolutionary model of cannabinoid oxidocyclase gene duplication and diversification. Most parsimious hypothesis based on microsynteny patterns shown in figure 4 and phylogenetic reconstruction shown in figure 3. Cannabinoid oxidocyclase genes are labeled in boldface. The nature of the hypothesized ancestral cannabinoid oxidocyclase remains unknown Vergara et al. We now know, however, that this perceived variation was due to the existence of additional gene lineages fig. In vitro functional studies suggest that these enzymes can produce multiple products and, therefore, perhaps the ancestral enzyme was promiscuous Zirpel et al. Nevertheless, given the relative recent divergence of cannabinoid oxidocyclase genes it may be possible to reconstruct an ancestral protein sequence for functional testing with reasonable accuracy. Earlier studies have assessed cannabinoid oxidocyclase gene copy number with the aim to link this to chemical variation McKernan et al. This is in line with recent findings based on a similar comparative genomics approach Hurgobin et al. Given that these sub clades may include a variable number of pseudogenes fig. Besides the divergence at the genomic level mentioned above, sequence variation within cannabinoid oxidocyclase gene sequences may help shed light on their evolutionary history. For example, subclade A4 pseudogenes are generally single copy but can be divided into two divergent types supplementary table S2 , Supplementary Material online; fig. These findings further corroborate our evolutionary interpretation of cannabinoid oxidocyclase gene duplication and divergence shown in figure 5 and suggest significant and consistent divergence between haplotypes containing CBDAS and THCAS. Thus, genomic divergence described above correlates with the prevalence of THCA and CBDA and, hence, perhaps with genetic origins of drug- versus fiber-type cultivars. Drug-type cultivars are considered to have originated in two different regions of the Himalayan foothills, while fiber-type cultivars are considered to have been developed independently in Europe and in East Asia Clarke and Merlin ; Clarke and Merlin The observed genetic variation may therefore be a consequence of divergence between these different geographic regions. S1 , Supplementary Material online. Accessions from Boseung province share three unique aa substitutions, the accession from Jecheon province in Korea that has five unique aa substitutions, and sequences from Cheungsam share ten unique aa substitutions Doh et al. This suggests that additional sampling throughout the native range of Cannabis is likely to reveal additional genetic variation. However, germplasm from regions of origin is scarce, especially when restricting samples to those that are compliant with international regulations such as the Nagoya-protocol. We sampled full-length predicted berberine bridge protein sequences from the Eurosid clade based on whole-genome assemblies of C. Some Cannabis and Humulus genes were found to be misannotated or lacking an annotation. These were manually corrected based on alignment with a closely related and correctly annotated genome sequence. In addition, we included daurichromenic acid synthase from Rhododendron dauricum accession BAZ Sequences of A. We mined available near chromosome-level genome assemblies of C. When necessary, structural annotations were manually modified based on nucleotide alignments with annotated genes with the highest identity. When genes comprised putative pseudogenes i. In addition, we compiled homologous nucleotide sequences available from ncbi databases the majority of which came from published studies supplementary table S3 , Supplementary Material online Sirikantaramas et al. Based on preliminary analyses, some sequences described by Cascini et al. Some sequences amplified from Moroccan hashish samples described by El Alaoui et al. These ambiguous and suspected chimeric sequences were excluded from final analyses. Optimal models of sequence evolution as determined using Modeltest-NG v. Gene trees were reconstructed in a Bayesian framework using MrBayes v 3. The first , generations were discarded as burnin. Within the berberine bridge gene family tree, clades were numbered in accordance with earlier classification Daniel et al. Within the cannabinoid oxidocyclase gene tree, clades and types were characterized based on unique nonsynonymous substitutions i. Because we found inconsistencies between the different genome assemblies in the ordering and orientation of sequences into scaffolds we considered genomic contigs only. Nucleotide-level alignments were generated by performing gapped extensions of high-scoring segment pairs using Lastz version 1. To keep tandem repeats we set —nochain. The —rdotplot output was used to generate alignment dotplots in R. For nucleotide level microsyntenic blocks of interest we further assessed microsynteny with genomic sequences from H. Supplementary data are available at Genome Biology and Evolution online. The sequence data underlying this article are available in the GenBank Nucleotide Database at www. The multiple-sequence alignments and associated gene trees are available in the 4TU. Google Scholar. Distribution of cannabinoid synthase genes in non- Cannabis organisms. J Cannabis Res. The draft genome and transcriptome of Cannabis sativa. Genome Biol. Berardini TZ , et al. Genesis 53 8 : — Bhattacharyya S , et al. Effect of cannabidiol on medial temporal, midbrain, and striatal dysfunction in people at clinical high risk of psychosis: a randomized clinical trial. JAMA Psychiatry. Brierley DI , et al. Chemotherapy-induced cachexia dysregulates hypothalamic and systemic lipoamines and is attenuated by cannabigerol. J Cachexia Sarcopenia Muscle. Cascini F , et al. Highly predictive genetic markers distinguish drug-type from fiber-type Cannabis sativa L. Plants 8 11 : Cannabis : evolution and ethnobotany. Google Preview. Cannabis domestication, breeding history, present-day genetic diversity, and future prospects. Daniel B , et al. The family of berberine bridge enzyme-like enzymes: a treasure-trove of oxidative reactions. Arch Biochem Biophys. Structure of a berberine bridge enzyme-like enzyme with an active site specific to the plant family Brassicaceae. PLoS One. Darriba D , et al. Mol Biol Evol. The inheritance of chemical phenotype in Cannabis sativa L. II : cannabigerol predominant plants. Euphytica : — Doh EJ , et al. DNA markers to discriminate Cannabis sativa L. Evid Based Complement Alternat Med. An experimental randomized study on the analgesic effects of pharmaceutical-grade cannabis in chronic pain patients with fibromyalgia. Pain 4 : — El Alaoui MA , et al. Phytochemistry of Cannabis sativa L. Progress in the Chemistry of Organic Natural Products, vol Cham: Springer International Publishing. Fellermeier M , Zenk MH. Prenylation of olivetolate by a hemp transferase yields cannabigerolic acid, the precursor of tetrahydrocannabinol. FEBS Lett. Ann Toxicol Anal. Gagne SJ , et al. Identification of olivetolic acid cyclase from Cannabis sativa reveals a unique catalytic route to plant polyketides. Nat Chem. Gaoni Y , Mechoulam R. Isolation, structure, and partial synthesis of an active constituent of hashish. J Am Chem Soc. A high-quality reference genome of wild Cannabis sativa. Hortic Res. SNP in potentially defunct tetrahydrocannabinolic acid synthase is a marker for cannabigerolic acid dominance in Cannabis sativa L. Genes 12 2 : Gofshteyn JS , et al. Cannabidiol as a potential treatment for febrile infection-related epilepsy syndrome FIRES in the acute and chronic phases. J Child Neurol. Grassa CJ , et al. A new Cannabis genome assembly associates elevated cannabidiol CBD with hemp introgressed into marijuana. New Phytol. Grimison P , et al. Phytocannabinoids: origins and biosynthesis. Trends Plant Sci. Chalcomoracin is a potent anticancer agent acting through triggering oxidative stress via a mitophagy- and paraptosis-dependent mechanism. Isolation and analysis of a gene bbe1 encoding the berberine bridge enzyme from the California poppy Eschscholzia californica. Plant Mol Biol. Cannabis : from cultivar to chemovar II—a metabolomics approach to cannabis classification. Cannabis Cannabinoid Res. Hurgobin B , et al. Recent advances in Cannabis sativa genomics research. Iijima M , et al. Identification and characterization of daurichromenic acid synthase active in anti-HIV biosynthesis. Plant Physiol. Born migrators: historical biogeography of the cosmopolitan family Cannabaceae. J Syst Evol. Development of loop-mediated isothermal amplification LAMP assay for rapid detection of Cannabis sativa. Biol Pharm Bull. Forensic Sci Int. Kovalchuk I , et al. The genomics of Cannabis and its close relatives. Annu Rev Plant Biol. Laverty KU , et al. Genome Res. Drivers of metabolic diversification: how dynamic genomic neighbourhoods generate new biosynthetic pathways in the Brassicaceae. Complete biosynthesis of cannabinoids and their unnatural analogues in yeast. Lynch RC , et al. Genomic and chemical diversity in cannabis. Qualitative and quantitative aspects of the inheritance of chemical phenotype in Cannabis. J Ind Hemp. McKernan K , et al. Cryptocurrencies and Zero Mode Wave guides: an unclouded path to a more contiguous Cannabis sativa L. Open Science Framework. Accessed June McKernan KJ , et al. Sequence and annotation of 42 Cannabis genomes reveals extensive copy number variation in cannabinoid synthesis and pathogen resistance genes. Models of Cannabis taxonomy, cultural bias, and conflicts between scientific and vernacular names. Bot Rev. A classification of endangered high-THC cannabis Cannabis sativa subsp. PhytoKeys : 81 — Mechoulam R. Plant cannabinoids: a neglected pharmacological treasure trove. Br J Pharmacol. Mechoulam R , Parker LA. The endocannabinoid system and the brain. Annu Rev Psychol. Genetics 1 : — In: ieeexplore. Enzymological evidence for cannabichromenic acid biosynthesis. J Nat Prod. Purification and characterization of cannabichromenic acid synthase from Cannabis sativa. Phytochemistry 49 6 : — Sequence heterogeneity of cannabidiolic- and tetrahydrocannabinolic acid-synthase in Cannabis sativa L. Phytochemistry : 57 — Padgitt-Cobb LK , et al. A phased, diploid assembly of the Cascade hop Humulus lupulus genome reveals patterns of selection and haplotype variation. Available from: Pertwee RG. Inverse agonism and neutral antagonism at cannabinoid CB1 receptors. Life Sci. Diversity and evolution of the repetitive genomic content in Cannabis sativa. BMC Genomics. Olivetol as product of a polyketide synthase in Cannabis sativa L. Plant Sci. Cannabinoid synthases and osmoprotective metabolites accumulate in the exudates of Cannabis sativa L. Comprehending and improving cannabis specialized metabolism in the systems biology era. Ronquist F , Huelsenbeck JP. MrBayes 3: Bayesian phylogenetic inference under mixed models. Bioinformatics 19 : — Analysis of cannabinoids in fiber hemp plant varieties Cannabis sativa L. Chromatographia 48 : — Sawler J , et al. The genetic structure of marijuana and hemp. PLoS One 10 : e Sirikantaramas S , et al. The gene controlling marijuana psychoactivity: molecular cloning and heterologous expression of Deltatetrahydrocnnabinolic acid synthasefrom Cannabis sativa L. J Biol Chem. Tetrahydrocannabinolic acid synthase, the enzyme controlling marijuana psychoactivity, is secreted into the storage cavity of the glandular trichomes. Plant Cell Physiol. Use of cannabidiol in anxiety and anxiety-related disorders. J Am Pharm Assoc. Small E. Dwarf germplasm: the key to giant Cannabis hempseed and cannabinoid crops. Genet Resour Crop Evol. Small E , Cronquist A. A practical and natural taxonomy for Cannabis. Taxon 25 4 : — The hexanoyl-CoA precursor for cannabinoid biosynthesis is formed by an acyl-activating enzyme in Cannabis sativa trichomes. Plant J. Suraev AS , et al. Cannabinoid therapies in the management of sleep disorders: A systematic review of preclinical and clinical studies. Sleep Med Rev. Tang H , et al. An improved genome release version Mt4. Purification and characterization of cannabidiolic-acid synthase from Cannabis sativa L. First direct evidence for the mechanism of. Phytocannabinoids in Cannabis sativa : recent studies on biosynthetic enzymes. Chem Biodivers. Taura F , et al. Cannabidiolic-acid synthase, the chemotype-determining enzyme in the fiber-type Cannabis sativa. Characterization of olivetol synthase, a polyketide synthase putatively involved in cannabinoid biosynthetic pathway. Toth JA , et al. Development and validation of genetic markers for sex and cannabinoid chemotype in Cannabis sativa L. GCB Bioenergy. Cannabichromene is a cannabinoid CB2 receptor agonist. Comparative genomics of the nonlegume Parasponia reveals insights into evolution of nitrogen-fixing rhizobium symbioses. Vergara D , et al. Genetic and genomic tools for Cannabis sativa. Gene copy number is associated with phytochemistry in Cannabis sativa. AoB Plants. Weiblen GD , et al. Gene duplication and divergence affecting drug content in Cannabis sativa. Welling MT , et al. A belated green revolution for cannabis: virtual genetic resources to fast-track cultivar development. Front Plant Sci. Wenger JP , et al. Validating a predictive model of cannabinoid inheritance with feral, clinical, and industrial Cannabis sativa. Am J Bot. Winkler A , et al. A concerted mechanism for berberine bridge enzyme. Nat Chem Biol. J Biotechnol. Oxford University Press is a department of the University of Oxford. It furthers the University's objective of excellence in research, scholarship, and education by publishing worldwide. 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What Do We Know About Medical Cannabis in Neurological Disorders and What Are the Next Steps?
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Thank you for visiting nature. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser or turn off compatibility mode in Internet Explorer. In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript. Analysis of over Cannabis samples quantified for terpene and cannabinoid content and genotyped for over , single nucleotide polymorphisms indicated that Sativa- and Indica-labelled samples were genetically indistinct on a genome-wide scale. Instead, we found that Cannabis labelling was associated with variation in a small number of terpenes whose concentrations are controlled by genetic variation at tandem arrays of terpene synthase genes. The vernacular labels Sativa and Indica not to be confused with the taxonomic names C. However, it is unclear whether these labels capture meaningful information about Cannabis genetic and chemical variation. Cannabis genomics research has thus far largely focused on the characterization of genes underlying the production of the cannabinoids cannabidiol CBD and tetrahydrocannabinol THC 5 , 6 , 7 , 8. However, Cannabis produces hundreds of aromatic terpenes that drive consumer preference and are frequently associated with Sativa and Indica labels 4 , 9. To date, various terpene synthase genes have been identified in Cannabis ; however, the genetic control of terpene variation across Cannabis cultivars remains largely unexplored 12 , 13 , 14 , Here we re-analysed samples of drug-type Cannabis that were previously quantified for 40 terpenes and cannabinoids using gas chromatography—mass spectrometry GC—MS 16 Supplementary Table 1 and Extended Data Fig. Principal component analysis PCA of the genomic data showed no clear clustering according to sample labels Fig. Sativa—Indica labels thus do not accurately reflect genetic relatedness, which is consistent with previous work 17 , In addition, we determined that pairs of samples with identical cultivar names for example, OG Kush were often as genetically and chemically distant from each other as pairs of samples with different names Extended Data Fig. The y axis shows the percent variance explained as PCs are added to linear models where the Sativa—Indica labelling scale is the dependent variable. Since the overall patterns of genetic and chemical relatedness could not fully account for the labels applied to Cannabis samples, we aimed to determine which individual chemicals were the strongest predictors of Sativa—Indica labelling. The strongest correlation was between Indica content and myrcene, whose concentration explained The sedative effect and earthy aroma attributed to high myrcene content are often reported by recreational users to be characteristic of Indica cultivars 10 , 24 , 25 , Hillig 27 found that these three sesquiterpenes were associated with plants from Afghanistan, which is considered the region of origin for Indica cultivars. The P values were Bonferroni-adjusted for multiple comparisons. The asterisks denote chemicals with tentative identifications. GWAS results are shown for chemicals highlighted in grey. The significance thresholds from the MLM are shown as horizontal dashed lines. Terpene synthase gene clusters are green. Previous chemical analyses of Cannabis have suggested that the distinction between Sativa and Indica is best explained by differences in the concentrations of specific monoterpenes and sesquiterpenes 19 , 28 , 29 , As a previous study suggested 31 , we hypothesize that Cannabis growers and breeders have been assigning labels to cultivars primarily on the basis of aroma profiles and purported effects, rather than genetic ancestry or overall chemical similarity. The primary differences between cultivars labelled as Sativa and Indica may thus be driven by a small set of genomic regions controlling the concentrations of a small number of contrasting aromas. To examine this, we conducted a genome-wide association study GWAS of the 40 chemicals examined here Supplementary Fig. We identified three regions of the Cannabis genome associated with the four terpenes most strongly associated with Sativa—Indica labelling Fig. The first SNP chr is located 6. The second SNP chr is Within this gene cluster are two sequences highly similar to the myrcene synthase gene, TPS3 refs. These observations suggest that myrcene synthesis is mediated by genetic variants at two independent terpene synthase gene clusters on chromosome 5. Our results demonstrate that the Sativa—Indica scale currently used to label Cannabis poorly captures overall genomic and metabolomic variation. Cannabis labelling is instead probably driven primarily by a small number of key terpenes whose concentrations contribute to the characteristic aromas commonly associated with Sativa and Indica and whose variation we genetically mapped to tandem arrays of terpene synthase genes on chromosomes 5 and 6. The samples come from a previous study of Cannabis chemotypes A total of samples were previously quantified for terpene and cannabinoid content, and we conduct a re-analysis of these data here. The chemical analyses of the samples are described in detail in ref. The supernatant was collected, and the process was repeated twice more on the pellet. The combined sample was analysed using an Agilent GC series Agilent Technologies equipped with a autosampler and a flame ionizing detector. Peaks from the sample chromatograms were manually integrated, and the peak area was recorded with correction for the internal standard peak area. Compounds without authentic standards are marked with an asterisk in the figures to indicate that they were tentative identifications. We re-assessed the compound identifications in Hazekamp et al. For example, in the case of the compound listed by Hazekamp et al. Thymoquinone, geraniol, thymol and carvacrol were removed because they were not present in any samples, and cineol was removed because it was present in only one sample. Pearson correlations were calculated between each pair of chemicals using the cor. According to previous work 33 , the samples analysed here were nearly all drug-type Cannabis that is, type I Extended Data Fig. Genotyping-by-sequencing libraries were prepared using the restriction enzyme ApeKI 34 , and the libraries were sequenced on two lanes of an Illumina Hi-Seq Illumina. Chromosomes were recoded for analyses to reflect the new chromosome numbering system. We used VCFtools v. Genotype imputation was performed using LinkImputeR 37 with a minor allele frequency threshold of 0. After imputation, samples remained. An additional 12 samples were removed because they had no phenotype data. This resulted in a final set of samples with both genetic and chemical data. The genetic similarity between samples was calculated as an inverse identity-by-state matrix generated in PLINK. The correlations between the matrices were computed using a Mantel test in R 32 by first reducing the chemical matrix to the samples with both chemical and genetic datasets. PCA was performed on the scaled genetic and chemical data using the prcomp function in R. To calculate the variance in labelling explained by the chemical and genetic data, linear models including the top ten PCs from the genetic data, the chemical data and both the chemical and genetic datasets together were performed. Pearson correlations between chemical concentration and the 1-to-5 Sativa—Indica scale were performed with the cor. A Bonferroni correction was applied to the P values from the correlation test between chemical concentration and the Sativa—Indica scale. We performed GWAS for 40 terpene and cannabinoid phenotypes, using both normalized and non-normalized data. Normalizing was conducted to generate values for a chemical concentration in a sample relative to the total abundance of its chemical class that is, monoterpene, sesquiterpene or cannabinoid in that sample. Quantile—quantile and Manhattan plots were created using the qq function in R. Genomic regions with significant GWAS hits were explored, and the physical locations of genes within these regions were retrieved using annotations from the CBDRx reference genome 8 in Geneious Prime v. Further information on research design is available in the Nature Research Reporting Summary linked to this article. The authors declare that the data supporting the findings are available within the paper. Lawler, A. Mountain high: oldest clear signs of pot use. Science , Naville, S. Bonini, S. Cannabis sativa : a comprehensive ethnopharmacological review of a medicinal plant with a long history. Guy, G. Models of Cannabis taxonomy, cultural bias, and conflicts between scientific and vernacular names. Laverty, K. Genome Res. McKernan, K. Sequence and annotation of 42 cannabis genomes reveals extensive copy number variation in cannabinoid synthesis and pathogen resistance genes. Vergara, D. Gene copy number is associated with phytochemistry in Cannabis sativa. AoB Plants 11 , plz Grassa, C. A new Cannabis genome assembly associates elevated cannabidiol CBD with hemp introgressed into marijuana. Gilbert, A. Consumer perceptions of strain differences in Cannabis aroma. Article Google Scholar. Russo, E. Taming THC: potential cannabis synergy and phytocannabinoid—terpenoid entourage effects. Koltai, H. Trends Plant Sci. Booth, J. Terpene synthases from Cannabis sativa. Zager, J. Gene networks underlying cannabinoid and terpenoid accumulation in Cannabis. Plant Physiol. Livingston, S. Cannabis glandular trichomes alter morphology and metabolite content during flower maturation. Plant J. Hazekamp, A. Cannabis: from cultivar to chemovar II—a metabolomics approach to cannabis classification. Cannabis Cannabinoid Res. Sawler, J. The genetic structure of marijuana and hemp. Lynch, R. Genomic and chemical diversity in Cannabis. Plant Sci. Henry, P. A single nucleotide polymorphism assay sheds light on the extent and distribution of genetic diversity, population structure and functional basis of key traits in cultivated North American cannabis. Cannabis Res. Schwabe, A. Genetic tools weed out misconceptions of strain reliability in Cannabis sativa : implications for a budding industry. Smith, C. The phytochemical diversity of commercial cannabis in the United States. Pearce, D. Discriminating the effects of Cannabis sativa and Cannabis indica : a web survey of medical cannabis users. Temple, L. Tetrahydrocannabinol—friend or foe? Hartsel, J. Gupta, R. Hillig, K. A chemotaxonomic analysis of terpenoid variation in Cannabis. Elzinga, S. Cannabinoids and terpenes as chemotaxonomic markers in cannabis. Casano, S. Variations in terpene profiles of different strains of Cannabis sativa L. Acta Hortic. Fischedick, J. Metabolic fingerprinting of Cannabis sativa L. Phytochemistry 71 , — Mudge, E. The terroir of cannabis: terpene metabolomics as a tool to understand Cannabis sativa selections. Planta Med. Small, E. The evolution of cannabinoid phenotypes in cannabis. Elshire, R. A robust, simple genotyping-by-sequencing GBS approach for high diversity species. Bradbury, P. Bioinformatics 23 , — Danecek, P. The variant call format and VCFtools. Bioinformatics 27 , — Money, D. LinkImputeR: user-guided genotype calling and imputation for non-model organisms. BMC Genomics 18 , Purcell, S. PLINK: a tool set for whole-genome association and population-based linkage analyses. Segura, V. An efficient multi-locus mixed-model approach for genome-wide association studies in structured populations. Gao, X. A multiple testing correction method for genetic association studies using correlated single nucleotide polymorphisms. Hu, Z. An integrated genotyping-by-sequencing polymorphism map for over 10, sorghum genotypes. Plant Genome 12 , Download references. We thank A. Hazekamp, M. Schranz and F. Becker for their contributions to this work. We thank C. Forney and T. Soomro for their assistance. You can also search for this author in PubMed Google Scholar. Correspondence to Sean Myles. Bedrocan funded this work, and R. The remaining authors declare no competing interests. Peer review information Nature Plants thanks Mahmoud A ElSohly, Andrea Mastinu and the other, anonymous, reviewer s for their contribution to the peer review of this work. Histograms of a pairwise chemical distances and b pairwise genetic distances among all pairs of samples. Vertical lines indicate the median distance between pairs of samples with the same name. Plot of genetic distance versus chemical distance between pairs of samples. The Mantel r statistic and p-value are reported. Heatmap displaying the Pearson correlation between the concentrations of the 40 terpenes and cannabinoids. Supplementary Table 1: Chemical concentrations and labels across Cannabis samples. The genomic coordinates and annotations, P value, R 2 value and nearby candidate genes are shown. Supplementary Table 3: A list of compound names identified by Hazekamp et al. Reprints and permissions. Watts, S. Cannabis labelling is associated with genetic variation in terpene synthase genes. Plants 7 , — Download citation. Received : 13 April Accepted : 03 August Published : 14 October Issue Date : October Anyone you share the following link with will be able to read this content:. Sorry, a shareable link is not currently available for this article. Provided by the Springer Nature SharedIt content-sharing initiative. Sign up for the Nature Briefing: Translational Research newsletter — top stories in biotechnology, drug discovery and pharma. Skip to main content Thank you for visiting nature. Download PDF. Subjects Genomics Natural variation in plants. Abstract Analysis of over Cannabis samples quantified for terpene and cannabinoid content and genotyped for over , single nucleotide polymorphisms indicated that Sativa- and Indica-labelled samples were genetically indistinct on a genome-wide scale. Genetic insights into agronomic and morphological traits of drug-type cannabis revealed by genome-wide association studies Article Open access 22 April Full size image. Methods Samples The samples come from a previous study of Cannabis chemotypes Gas chromatography A total of samples were previously quantified for terpene and cannabinoid content, and we conduct a re-analysis of these data here. Genome-wide association We performed GWAS for 40 terpene and cannabinoid phenotypes, using both normalized and non-normalized data. Reporting Summary Further information on research design is available in the Nature Research Reporting Summary linked to this article. Data availability The authors declare that the data supporting the findings are available within the paper. References Lawler, A. Article Google Scholar Russo, E. Article Google Scholar Zager, J. Article Google Scholar Lynch, R. Article Google Scholar Henry, P. Article Google Scholar Schwabe, A. Article Google Scholar Smith, C. Article Google Scholar Temple, L. Article Google Scholar Casano, S. Article Google Scholar Fischedick, J. Article Google Scholar Purcell, S. Article Google Scholar Hu, Z. Article Google Scholar Download references. Acknowledgements We thank A. View author publications. Ethics declarations Competing interests R. Additional information Peer review information Nature Plants thanks Mahmoud A ElSohly, Andrea Mastinu and the other, anonymous, reviewer s for their contribution to the peer review of this work. Extended data. Extended Data Fig. Supplementary information Supplementary Information Supplementary Figs. Reporting Summary. Supplementary Tables Supplementary Table 1: Chemical concentrations and labels across Cannabis samples. About this article. Cite this article Watts, S. Copy to clipboard. Search Search articles by subject, keyword or author. Show results from All journals This journal. Advanced search. Close banner Close. Email address Sign up. Get what matters in translational research, free to your inbox weekly. Sign up for Nature Briefing: Translational Research.
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