Glandular trichomes are specialized anatomical structures that accumulate secretions with important biological roles in plant-environment interactions. These secretions also have commercial uses in the flavor, fragrance, and pharmaceutical industries. The capitate-stalked glandular trichomes of Cannabis sativa (cannabis), situated on the surfaces of the bracts of the female flowers, are the primary site for the biosynthesis and storage of resins rich in cannabinoids and terpenoids. In this study, we profiled nine commercial cannabis strains with purportedly different attributes, such as taste, color, smell, and genetic origin. Glandular trichomes were isolated from each of these strains, and cell type-specific transcriptome data sets were acquired. Cannabinoids and terpenoids were quantified in flower buds. Statistical analyses indicated that these data sets enable the high-resolution differentiation of strains by providing complementary information. Integrative analyses revealed a coexpression network of genes involved in the biosynthesis of both cannabinoids and terpenoids from imported precursors. Terpene synthase genes involved in the biosynthesis of the major monoterpenes and sesquiterpenes routinely assayed by cannabis testing laboratories were identified and functionally evaluated. In addition to cloning variants of previously characterized genes, specifically CsTPS14CT [(−)-limonene synthase] and CsTPS15CT (β-myrcene synthase), we functionally evaluated genes that encode enzymes with activities not previously described in cannabis, namely CsTPS18VF and CsTPS19BL (nerolidol/linalool synthases), CsTPS16CC (germacrene B synthase), and CsTPS20CT (hedycaryol synthase). This study lays the groundwork for developing a better understanding of the complex chemistry and biochemistry underlying resin accumulation across commercial cannabis strains.

Gene Networks Underlying Cannabinoid and Terpenoid Accumulation in Cannabis

Plant pathogens infecting marijuana (Cannabis sativa L.) plants reduce growth of the crop by affecting the roots, crown and foliage. In addition, fungi (molds) that colonize the inflorescences (buds) during development or after harvest, and which colonize internal tissues as endophytes, can reduce product quality. The pathogens and molds that affect C. sativa grown hydroponically indoors (in environmentally controlled growth rooms and greenhouses) and field-grown plants were studied over multiple years of sampling. A PCR-based assay using primers for the internal transcribed spacer region (ITS) of ribosomal DNA confirmed identity of the cultures. Root-infecting pathogens included Fusarium oxysporum, F. solani, F. brachygibbosum, Pythium dissotocum, P. myriotylum and P. aphanidermatum, which caused root browning, discoloration of the crown and pith tissues, stunting and yellowing of plants, and in some instances, plant death. On the foliage, powdery mildew, caused by Golovinomyces cichoracearum, was the major pathogen observed. On inflorescences, penicillium bud rot (caused by P. olsonii and P. copticola), botrytis bud rot (B. cinerea) and fusarium bud rot (F. solani, F. oxysporum) were present to varying extents. Endophytic fungi present in crown, stem and petiole tissues included soil-colonizing and cellulolytic fungi, such as species of Chaetomium, Trametes, Trichoderma, Penicillium and Fusarium. Analysis of air samples in indoor growing environments revealed that species of Penicillium, Cladosporium, Aspergillus, Fusarium, Beauvaria, and Trichoderma were present. The latter two species were the result of the application of biocontrol products for control of insects and diseases, respectively. Fungal communities present in unpasteurized coconut fibre (coco) growing medium are potential sources of mold contamination on cannabis plants. Swabs taken from greenhouse-grown and indoor buds pre- and post-harvest revealed the presence of Cladosporium and up to five species of Penicillium, as well as low levels of Alternaria species. Mechanical trimming of buds caused an increase in the frequency of Penicilllium species, presumably by providing entry points through wounds or spreading endophytes from pith tissues. Aerial distribution of pathogen inoculum and mold spores and dissemination through vegetative propagation, are important methods of spread, and entry through wound sites on roots, stems and bud tissues facilitates pathogen establishment on cannabis plants.

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Pathogens and Molds Affecting Production and Quality of Cannabis sativa L.

Recent advances in cannabis biotechnology

Flower buds of Cannabis sativa develop as inflorescences (buds) which are harvested and dried prior to sale. The extent to which fungal plant pathogens can colonize the buds prior to harvest has no...

https://www.tandfonline.com/doi/pdf/10.1080/07060661.2018.1535467

Flower and foliage-infecting pathogens of marijuana (Cannabis sativa L.) plants

Cultivation of cannabis plants (Cannabis sativa L., marijuana) has taken place worldwide for centuries. In Canada, legalization of cannabis in October 2018 for the medicinal and recreational markets has spurned interest in large-scale growing. This increased production has seen a rise in the incidence and severity of plant pathogens, causing a range of previously unreported diseases. The objective of this review is to highlight the important diseases currently affecting the cannabis and hemp industries in North America and to discuss various mitigation strategies. Progress in molecular diagnostics for pathogen identification and determining inoculum sources and methods of pathogen spread have provided useful insights. Sustainable disease management approaches include establishing clean planting stock, modifying environmental conditions to reduce pathogen development, implementing sanitation measures, and applying fungal and bacterial biological control agents. Fungicides are not currently registered for use and hence there are no published data on their efficacy. The greatest challenge remains in reducing microbial loads (colony-forming units) on harvested inflorescences (buds). Contaminating microbes may be introduced during the cultivation and postharvest phases, or constitute resident endophytes. Failure to achieve a minimum threshold of microbes deemed to be safe for utilization of cannabis products can arise from conventional and organic cultivation methods, or following applications of beneficial biocontrol agents. The current regulatory process for approval of cannabis products presents a challenge to producers utilizing biological control agents for disease management. © 2021 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

https://onlinelibrary.wiley.com/doi/pdf/10.1002/ps.6307

Emerging Diseases of Cannabis sativa and Sustainable Management.

Yellowing and wilting symptoms on field-grown Cannabis sativa (cannabis) plants followed by total plant collapse under conditions of extreme hot weather were observed in northern California in 2017. The crown regions of affected plants were dark and sunken and internal tissue discolouration extended 10–15 cm above the soil surface. Isolations made from the pith, vascular and cortical tissues in the crown region yielded Fusarium oxysporum (40% frequency), F. brachygibbosum (28% frequency), Pythium aphanidermatum (22% frequency), Fusarium solani and F. equiseti (5% frequency each). Pathogenicity tests were conducted on rooted plantlets to establish the extent of root and crown decay, as well as on mature stems to determine the extent of stem tissue colonization caused by these species. Extensive reduction in root length was caused by F. solani, F. oxysporum, F. brachygibbosum and P. aphanidermatum and wounding significantly enhanced disease development. Stem tissue colonization by these pathogens at...

https://www.tandfonline.com/doi/pdf/10.1080/07060661.2018.1535470

Root and crown rot pathogens causing wilt symptoms on field-grown marijuana (Cannabis sativa L.) plants

Cannabis sativa L represents plants cultivated for their psychoactive and medicinal properties (marijuana) or as a source of fibre, seed and oil (hemp) Breeding and selection efforts have produced marijuana genotypes (strains) with a range of levels of the cannabinoid Δ9-tetrahydrocannabinolic acid (THCA) and other non-psychoactive cannabinoids, eg cannabidiolic acid (CBDA) Hemp lines have been bred for high fibre content and seed production and have low/no THCA There are currently hundreds of marijuana strains which differ in THCA:CBDA ratios, growth characteristics, morphological features, THCA and CBDA contents, disease resistance, as well as overall medicinal value The extent of genetic diversity among these marijuana strains, as well as between marijuana and hemp, has been studied using a range of molecular approaches The results from these studies have demonstrated that considerable genetic diversity exists among marijuana as well as hemp strains Using ISSR markers, we have shown that distinct DNA banding patterns can allow for the initial discrimination between many of the strains tested, and provide an insight into the possible genetic relationships among strains Some strains, eg ‘Jack’, ‘Super Sour Skunk’, ‘Jilly Bean’, exhibited unique patterns that can be used to develop strain-specific DNA fingerprints In addition, a number of “landraces” and strains originating from remote geographic locations, showed unique and distinct ISSR patterns and morphologies A very high degree of genetic diversity was exhibited among the strains studied Additional molecular studies, including DNA sequencing approaches, should provide more insight into the genetic relationships that exist within strains of a complex plant species and could augment future breeding efforts for genetic improvement of C sativa

Sarah Chen

Papers

Root and crown rot pathogens causing wilt symptoms on field-grown marijuana (Cannabis sativa L.) plants

Assessing Genetic Diversity in Cannabis sativa Using Molecular Approaches