Cannabis Varieties Research Articles

Comprehensive Phenotypic Characterization of Diverse Drug-Type Cannabis Varieties from the Canadian Legal Market.

Cannabis (Cannabis sativa L.) stands as a historically significant and culturally important plant, embodying economic, social, and medicinal relevance for human societies. However, years of prohibition and stigmatization have hindered the cannabis research community, which is hugely undersized and suffers from a scarcity of understanding of cannabis genetics and how key traits are expressed or inherited. In this study, we conducted a comprehensive phenotypic characterization of 176 drug-type cannabis accessions, representative of Canada's legal market. We assessed germination methods, evaluated various traits including agronomic, morphological, and cannabinoid profiles, and uncovered significant variation within this population. Notably, the yield displayed a negative correlation with maturity-related traits but a positive correlation with the fresh biomass. Additionally, the potential THC content showed a positive correlation with maturity-related traits but a negative correlation with the yield. Significant differences were observed between the plants derived from regular female seeds and feminized seeds, as well as between the plants derived from cuttings and seeds for different traits. This study advances our understanding of cannabis cultivation, offering insights into germination practices, agronomic traits, morphological characteristics, and biochemical diversity. These findings establish a foundation for precise breeding and cultivar development, enhancing cannabis's potential in the legal market.

Урожайность и содержание белка в семенах сортов среднерусской однодомной конопли при разных нормах высева в Среднем Предуралье

The article presents the results of scientific research for 2020-2022 on the study of the content and harvesting of protein with the yield of cannabis varieties depending on the seeding rate in the soil and meteorological conditions of the Middle Urals. The soil of the experimental plots in the years of laying the experiments differed in the agrochemical characteristics of the arable layer: low and medium humus content, high and elevated – mobile phosphorus, elevated and very high – mobile potassium. The soil solution had a strongly acidic, medium acidic and close to neutral reaction. On sod-medium-podzolic medium loamy soil, the Vera variety stood out in terms of seed yield, providing seed productivity of 71 g/m2 in 2020, 193 g/m2 in 2021 and 128 g/m2 in 2022 with a crude protein content of 21.6%, 20.2%, 22.4%, respectively, by year. On average, over the years of research, the collection of raw protein with a yield of 291 and 303 kg/ha of the Nadezhda and Vera varieties had 1.2 million units/ha at the seeding rate, Surskaya hemp formed more raw protein in a yield of 273 kg/ha at a seeding rate of 0.8 million units/ha. The seeding rate of 0.4 million units/ha provided the highest 22.0–25.7% crude protein content in hemp seeds. Keywords: HEMP, VARIETY, SEEDING RATE, CRUDE PROTEIN, YIELD, SEEDS

The influence of agricultural practices on photosynthetic activity and productivity of a new variety of cannabis in the Middle Volga region

Relevance. In 2023, a new variety of single-domed drug-free cannabis, Lyudmila, of the fibrous direction of use, passed the State Examination and was approved for accreditation for admis-sion to use on the territory of the Russian Federation. In this regard, research on the development of elements of cultivation technology in specific soil and climatic conditions is of scientific and practical interest.Methods. The scientific experiment is based on methodological guidelines for registration tests offungicides in agriculture and methodological guidelines for conducting field and vegetation tests with cannabis. Indicators of photosynthetic activity of plants in crops were determined by the method of A.A. Nichiporovich. Mathematical processing of experimental data was carried out by the method of dispersion analysis according to B.A. Dospekhov.Results. Foliar top dressing contributed to an increase in the assimilation surface of the leaves in the variants with the treatment of plants for vegetation with preparations: Izagri Phosphorus — from 111.0 to 155.7 thousand m2/ha, Izagri Vita — from 112.2 to 139.7 thousand m2/ha, Humate +7 — from 135.9 to 148.4 thousand m2/ha (with 101.1–114.3 thousand m2/ha under control). The high yield of stems was obtained when sowing with a seeding rate of 3 million germinating seeds per hectare — 15.67 t/ha, with a decrease in the seeding rate to 2.5 and 2.0 million germinating seeds per hectare, the yield of stems decreased to 14.48–12.98 t/ha. Foliar treatments are effective, the yield of stems on the treated variants is higher than the control by 0.36–0.9 t/ha. The highest seed yield was obtained when sowing with a seeding rate of 2.5 million germinating seeds per hectare, on average 0.98 t/ha.

Cannabis sativa: origin and history, glandular trichome development, and cannabinoid biosynthesis.

Is Cannabis a boon or bane? Cannabis sativa has long been a versatile crop for fiber extraction (industrial hemp), traditional Chinese medicine (hemp seeds), and recreational drugs (marijuana). Cannabis faced global prohibition in the twentieth century because of the psychoactive properties of ∆9-tetrahydrocannabinol; however, recently, the perspective has changed with the recognition of additional therapeutic values, particularly the pharmacological potential of cannabidiol. A comprehensive understanding of the underlying mechanism of cannabinoid biosynthesis is necessary to cultivate and promote globally the medicinal application of Cannabis resources. Here, we comprehensively review the historical usage of Cannabis, biosynthesis of trichome-specific cannabinoids, regulatory network of trichome development, and synthetic biology of cannabinoids. This review provides valuable insights into the efficient biosynthesis and green production of cannabinoids, and the development and utilization of novel Cannabis varieties.

Features of forming the productivity of modern hemp varieties using organic cultivation technology

Seed hemp can be used to produce products and restore soil fertility. The adaptive potential of cannabis varieties can be implemented by optimising their cultivation technologies. The purpose of this study was to substantiate the features of forming the productivity of cannabis varieties adapted to organic cultivation technologies. Field studies were conducted during 2019-2021 on leached chernozems. Research methods: field, laboratory, statistical and comparative calculation. The regularities of yield formation of hemp straws, fibre, and seeds depending on the genetic potential of varieties and cultivation technology were investigated. The suitability of five hemp varieties for organic cultivation was evaluated. It was established that the introduction of organic cultivation technology has a positive effect on crop productivity. With organic cultivation of Hliana and Hloba varieties, the increase in the yield of straw (0.07-0.11 t/ha) was statistically insignificant compared to inorganic technology. Cultivation of Zolotoniski 15 and Sula varieties using organic technologies increased the yield of hemp straw by 0.14 t/ha. The Lara variety showed an insignificant decrease in the straw yield. This confirmed the theory that the introduction of organic cultivation of hemp will not lead to a decrease in yields. Organic technologies for growing hemp contributed to an increase in their yield compared to the traditional one, provided that varieties were selected correctly. The best suitability for organic cultivation in terms of fibre yield was recorded in the Lara variety. The obtained results will be used to improve and introduce organic technologies for growing adapted hemp varieties, depending on the purpose of fibre or seeds

Arbuscular Mycorrhizal Symbiosis in Two Cannabis Varieties (Khardala and Critical) in Morocco

Arbuscular Mycorrhizal Symbiosis in Two Cannabis Varieties (Khardala and Critical) in Morocco

Antifungal and antibacterial activities of Cannabis sativa L. resins.

Ethnopharmacological relevanceCannabis sativa L. (Cannabaceae) is a plant native to Eastern Asia spread throughout the world because of its medicinal properties. Despite being used for thousands of years as a palliative therapeutic agent for many pathologies, in many countries research on its effects and properties could only be carried out in recent years, after its legalization. Aims of the studyIncreasing resistance to traditional antimicrobial agents demands finding new strategies to fight against microbial infections in medical therapy and agricultural activities. Upon legalization in many countries, Cannabis sativa is gaining attention as a new source of active components, and the evidence for new applications of these compounds is constantly increasing. MethodsExtracts from five different varieties ofCannabis sativa were performed and their cannabinoids and terpenes profiles were determined by liquid and gas chromatography. Antimicrobial and antifungal activities against Gram (+) and Gram (−) bacteria, yeast and phytopathogen fungus were measured. To analyze a possible action mechanism, cell viability of bacteria and yeast was assessed by propidium iodide stain. ResultsCannabis varieties were grouped into chemotype I and II as a consequence of their cannabidiol (CBD) or tetrahydrocannabinol (THC) content. The terpenes profile was different in quantity and quality among varieties, with (−)b-pinene, b-myrcene, p-cymene and b-caryophyllene being present in all plants. All cannabis varieties were effective to different degree against Gram (+) and Gram (−) bacteria as well as on spore germination and vegetative development of phytopathogenic fungi. These effects were not correlated to the content of major cannabinoids such as CBD or THC, but with the presence of a complex terpenes profile. The effectiveness of the extracts allowed to reduce the necessary doses of a widely used commercial antifungal to prevent the development of fungal spores. ConclusionAll the extracts of the analysed cannabis varieties showed antibacterial and antifungal activities. In addition, plants belonging to the same chemotype showed different antimicrobial activity, demonstrating that the classification of cannabis strains based solely on THC and CBD content is not sufficient to justify their biological activities and that other compounds present in the extracts are involved in their action against pathogens. Cannabis extracts act in synergy with chemical fungicides, allowing to reduce its doses.

Analysis of a Professional Development Event on Cannabis Education for Cannabis Educators

Cannabis is one of the fastest growing industries in the US. The unprecedented expansion of the industry has been influenced by the changing regulatory landscape on industrial hemp, medical cannabis and adult-use cannabis. The wide variety of cannabis- and hemp-related jobs, from agricultural production to product sales, require a skilled workforce. Post-secondary institutions have recognized the gap in workforce training and have begun to offer educational credentials in cannabis ranging from certificates to graduate degrees. As the cannabis education sector is built, professional development on cannabis education for post-secondary educators can assist in the creation and teaching of high-quality academic programs. A survey was used to evaluate the inaugural Cannabis Curriculum Convening, a pioneering professional development event on cannabis education. Our findings show the professional development positively impacted educators’ knowledge and confidence toward integrating cannabis curriculum. Educators’ future needs for professional development (curriculum/program development and pedagogical content knowledge) were assessed. Findings show that educators desire professional development on forming cannabis industry partnerships and providing non-formal cannabis education. The most in-demand pedagogical content knowledge topics were medical cannabis, cannabis history, and cannabis regulation, testing, and compliance. We hope that our recommendations will inform current and future professional development in cannabis education.

Valorization of cannabis green waste to cellulose nanomaterials via phosphoric acid hydrolysis

The recent increases in Cannabis sativa (cannabis) production, both in industrial hemp and recreational varieties, has highlighted the need for low-impact and sustainable practices for this expanding industry. Green waste of cannabis, consisting of the vegetative components like plant stalks, offers an unexplored opportunity for valorization through biomass optimization. Like hemp, recreational cannabis varieties contain two high cellulose fiber types: bast fiber (phloem tissue) and hurd fiber (xylem tissue). Using hemp as a model for recreational cultivars, this study demonstrates the ability to isolate cellulose nanomaterials from both bast and hurd fiber through phosphoric acid hydrolysis. Scanning electron microscopy (EM) shows that sodium hydroxide treatment is required for removing all non-cellulose surface components from hurd fiber. Comparison of the two fiber types with transmission EM reveals phosphoric acid-hydrolyzed bast fibers produce cellulose nanocrystals (CNC), while hurd fibers are more prone to producing a mixture of cellulose nanocrystals and nanofibril-like particles (CNF) without chemical pretreatment. Adding hydrogen peroxide to the sodium hydroxide pretreatment decreased the variation in CNC sizes resulting from both bast and hurd. The findings of this study provide several avenues for valorization of green waste, while aiming to minimize the techno-economic impact of pretreatments.

CANNABIS AND MALE FERTILITY : A SYSTEMATIC REVIEW

Cannabis is a class of psychoactive substances derived from Cannabis sativa or Cannabis indica that can induce euphoria in the user. Cannabinoids are the term used to refer to the active component of cannabis, which contains 400 active compounds. DSM criterion 5 characterizes cannabis use disorder. Cannabis is the most commonly used illicit substance worldwide. The annual prevalence of cannabis abuse ranges from 2.9% to 4.3% of the global population aged 15 to 64. Cannabis sativa, Cannabis indica, and Cannabis ruderalis are the three varieties of cannabis. These three cannabis varieties contain varying amounts of tetrahydrocannabinol (THC). The THC content of Charas and hashish ranges between 7% and 14%. Cannabis and Sinsemilla are found in female shoots with an average THC concentration of 4-5% (rarely >7%). Low-grade Bhang preparations are derived from the 1% THC-containing residue of desiccated plants. Hashish oil, a concentrated liquid extracted from hashish distilleries, contains between 15 and 70 percent THC. Semen parameters provide the strongest evidence of cannabis-induced alterations in male fertility. Cannabis reduces sperm count and concentration, induces abnormalities in sperm morphology, reduces sperm motility and viability, and inhibits capacitation and fertilizing capacity, according to research.

Intelligent framework for cannabis classification using visualization of gas chromatography/mass spectrometry data and transfer learning

Introduction: Gas chromatography combined with mass spectrometry (GC/MS) is popular analytical instrumentation for chemical separation and identification. A novel framework for chemical forensics based on the visualization of GC/MS data and transfer learning is proposed.Methods: To evaluate the framework, 228 GC/MS data collected from two standard cannabis varieties, i.e., hemp and marijuana, were utilized. By processing the raw GC/MS data, analytical features, including retention times, mass-to-charge ratios, intensities, and summed ion mass spectra, were successfully transformed into two types of image representations. The GC/MS data transformed images were fed into a pre-trained convolutional neural network (CNN) to develop intelligent classifiers for the sample classification tasks. The effectiveness of several hyper-parameters for improving classification performance was investigated during transfer learning.Results: The proposed analytical workflow could classify hemp and marijuana with 97% accuracy. Furthermore, the transfer-learning-based classifiers were established without requiring big data sets and peak alignment.Discussion: The potential application of the new artificial intelligence (AI)-powered framework for chemical forensics using GC/MS data has been demonstrated. This framework provides unique opportunities for classifying various types of physical evidence using chromatography and mass spectrometry signals.

Recent Research on Cannabis sativa L.: Phytochemistry, New Matrices, Cultivation Techniques, and Recent Updates on Its Brain-Related Effects (2018-2023).

Cannabis sativa L. is a plant that humankind has been using for millennia. The basis of its widespread utilization is its adaptability to so many different climatic conditions, with easy cultivability in numerous diverse environments. Because of its variegate phytochemistry, C. sativa has been used in many sectors, although the discovery of the presence in the plant of several psychotropic substances (e.g., Δ9-tetrahydrocannabinol, THC) caused a drastic reduction of its cultivation and use together with its official ban from pharmacopeias. Fortunately, the discovery of Cannabis varieties with low content of THC as well as the biotechnological development of new clones rich in many phytochemical components endorsed with peculiar and many important bioactivities has demanded the reassessment of these species, the study and use of which are currently experiencing new and important developments. In this review we focus our attention on the phytochemistry, new matrices, suitable agronomic techniques, and new biological activities developed in the five last years.

Lessons from 20 years of medical cannabis use in Canada.

Canada was one of the first countries to regulate the medical use of cannabis. However, literature on Canada's medical cannabis program is limited. We use administrative data from the medical cannabis program, and licensed cannabis vendor catalog data to describe a) the participation of patients, physicians, and cannabis vendors in the program from its inception in 1999 to 2021, and b) trends in medical cannabis consumption, prices and potency. We also use national surveys conducted over the last several decades to estimate trends in regular cannabis use (medical or otherwise) and how it changed during the medical cannabis access regimes. In 2001, the Canadian government granted access to those with physician-documented evidence of a severe health problem that could not be managed using conventional therapies. Most patients accessed cannabis grown under a personal production license. By 2013, authorized daily cannabis dosages were very high. In 2014, the government, concerned over illegal diversion, required that cannabis be purchased from a licensed commercial grower; personal production was banned. Physicians were given responsibility for authorizing patient access. To fill the regulatory void, the physician regulatory bodies in Canada imposed their own prescribing restrictions. After these changes, the number of physicians who were willing to support patient cannabis use markedly decline but the number of patients participating in the program sharply increased. Medical cannabis use varied by province-rates were generally lower in provinces with stricter regulations on physician cannabis prescribing. Most varieties of cannabis oil available for sale are now high in CBD and low in THC. Dry cannabis varieties, conversely, tend to be high in THC and low in CBD. Inflation adjusted prices of most varieties of medical cannabis have declined over time. We find that rates of daily cannabis use (medical or otherwise) increased markedly after the 2014 policy regime. The fraction of Canadians using cannabis daily increased again after the 2018 legalization of recreational cannabis; at the same time, participation in the medical access program declined. The implications for patient health outcomes of changes in the medical cannabis program and legalization of recreational use remains an important area for future research.

Enantioseparation of chiral phytocannabinoids in medicinal cannabis

The evaluation of the chiral composition of phytocannabinoids in the cannabis plant is particularly important as the pharmacological effects of the (+) and (-) enantiomers of these compounds are completely different. Chromatographic attempts to assess the presence of the minor (+) enantiomers of the main phytocannabinoids, cannabidiolic acid (CBDA) and trans-Δ9-tetrahydrocannabinolic acid (trans-Δ9-THCA), were carried out on heated plant extracts for the determination of the corresponding decarboxylated species, cannabidiol (CBD) and trans-Δ9-tetrahydrocannabinol (trans-Δ9-THC), respectively. This process produces an altered phytocannabinoid composition with several new and unknown decomposition products. The present work reports for the first time the stereoselective synthesis of the pure (+) enantiomers of the main phytocannabinoids, trans-CBDA, trans-Δ9-THCA, trans-CBD and trans-Δ9-THC, and the development and optimization of an achiral-chiral liquid chromatography method coupled to UV and high-resolution mass spectrometry detection in reversed phase conditions (RP-HPLC-UV-HRMS) for the isolation of the single compounds and evaluation of their actual enantiomeric composition in plant. The isolation of the peaks with the achiral stationary phase ensured the absence of interferences that could potentially co-elute with the analytes of interest in the chiral analysis. The method applied to the Italian medicinal cannabis variety FM2 revealed no trace of the (+) enantiomers for all phytocannabinoids under investigation before and after decarboxylation, thus suggesting that the extraction procedure does not lead to an inversion of configuration.

Moving Away from 12:12; the Effect of Different Photoperiods on Biomass Yield and Cannabinoids in Medicinal Cannabis

The standard practice to initiate flowering in medicinal cannabis involves reducing the photoperiod from a long-day period to an equal duration cycle of 12 h light (12L)/12 h dark (12D). This method reflects the short-day flowering dependence of many cannabis varieties but may not be optimal for all. We sought to identify the effect of nine different flowering photoperiod treatments on the biomass yield and cannabinoid concentration of three medicinal cannabis varieties. The first, "Cannatonic", was a high cannabidiol (CBD)-accumulating line, whereas the other two, "Northern Lights" and "Hindu Kush", were high Δ9-tetrahydrocannabinol (THC) accumulators. The nine treatments tested, following 18 days under 18 h light/6 h dark following cloning and propagation included a standard 12L:12D period, a shortened period of 10L:14D, and a lengthened period of 14L:10D. The other six treatments started in one of the aforementioned and then 28 days later (mid-way through flowering) were switched to one of the other treatments, thus causing either an increase of 2 or 4 h, or a decrease of 2 or 4 h. Measured parameters included the timing of reproductive development; the dry weight flower yield; and the % dry weight of the main target cannabinoids, CBD and THC, from which the total g cannabinoid per plant was calculated. Flower biomass yields were highest for all lines when treatments started with 14L:10D; however, in the two THC lines, a static 14L:10D photoperiod caused a significant decline in THC concentration. Conversely, in Cannatonic, all treatments starting with 14L:10D led to a significant increase in the CBD concentration, which led to a 50-100% increase in total CBD yield. The results show that the assumption that a 12L:12D photoperiod is optimal for all lines is incorrect as, in some lines, yields can be greatly increased by a lengthened light period during flowering.

Contemplating cannabis? The complex relationship between cannabinoids and hepatic metabolism resulting in the potential for drug-drug interactions.

The majority of states have fully legalized the use of medical cannabis (MC), and nearly all other states allow limited access to cannabidiol (CBD), a non-intoxicating constituent of cannabis often touted for a range of therapeutic indications. Further, the Agricultural Improvement Act of 2018 legalized hemp-derived products in all 50 states; typically high in CBD, these products are derived from cannabis varieties containing ≤0.3% delta-9-tetrahydrocannabinol (THC) by weight. The recent "green rush" has resulted in a striking increase in cannabis use among patients and consumers who often use a wide variety of novel product types, each with a unique blend of cannabinoid constituents. Importantly, however, several cannabinoids have the potential to cause drug-drug interactions (DDI) with other medications, primarily due to their involvement with the hepatic cytochrome P450 (CYP450) system. This article examines the potential for individual cannabinoids, particularly CBD, to interact with the hepatic metabolic system, which is concerning given its involvement in the metabolism of commonly-prescribed medications. CBD and other cannabinoids are metabolized extensively by the CYP450 system, and also inhibit many of these enzymes, potentially leading to variable serum levels of other medications, as well as variable levels of cannabinoids when other medications modify the system. As access and interest in cannabinoid-based products continues to increase, critical questions remain unanswered regarding their safety. The complex relationship between cannabinoids and the hepatic metabolic system, including common potential DDI resulting from cannabinoid exposure, are explored along with the clinical significance of these potential interactions and monitoring or mitigation strategies.

Can we interrogate public databases to fill critical gaps in mental health epidemiology? Testing the association between cannabis and psychosis in the UK as an example.

The psychoactive properties of cannabis have been known forever. Since 1987, several prospective studies have suggested an increased risk of psychosis among cannabis users, with alternative explanations failing to account for such an effect. A cause-effect relationship has thus been implied. Further evidence has indicated that there is a dose-response relationship, and high-potency cannabis varieties confer the greatest risk of psychosis. As cannabis use has become more common over the last decades, one would expect a related increase in the number of schizophrenia cases. However, evidence in this regard remains equivocal for several reasons, including relying on databases that are not primarily designed to address such question and the issue that solid information regarding the incidence of schizophrenia is a relatively recent acquisition. Recent years have seen the development of online web publications, such as Google Trends and "Our World in Data", where data are explorable and interactable for tracking and comparing trends over specific periods and world regions. By using such databases, we believe that the question whether changes in cannabis use are associated with changes in schizophrenia rates can be answered, at least partly. Therefore, we tested these tools by evaluating trends in cannabis use and both cases and prevalence of schizophrenia in the United Kingdom, one of the countries where the incident rates for psychotic disorder have been suggested to be particularly increased by cannabis consumption. Crossing data from these tools revealed that interest in cannabis has been growing at the country level for over 10 years, with a parallel overlapping raise in psychosis cases and prevalence. Following up on this example, let us think of how many public health opportunities these public resources may offer. The question now is whether public health interventions for the benefit of the general population will follow suit.

Cannabis sativa: Botany, Cross Pollination and Plant Breeding Problems

This review paper highlights about the Cannabis botany, problems of cross pollination, lack of Germplasm collection and absence of Genebank facilities have been discussed. Cannabis is a dioecious species, meaning that male and female flowers are borne on separate plants. The risk of cross-pollination in Cannabis is one of the unique major challenge in Cannabis industry. More research can be conducted to assess the risk of cross-pollination in Cannabis and policy should be created to mitigate that risk. Cannabis research work remains years behind than other crops because of the long legacy of prohibition and stigmatization. However, lack of public germplasm repositories remains an unresolved problem in the Cannabis industry. Therefore, accessible germplasm resources are crucial for long-term economic viability, preserving genetic diversity, breeding, innovation, and long-term sustainability of the Cannabis crop. Germplasm accessions are the building blocks for breeding of new cultivars. However, there are no public seed or Clonal gene banks for Medical Cannabis sativa (marijuana or drug type). On the other hand there are some public hemp collections. Despite its economic, medicinal, and societal importance, Medical Cannabis sativa (marijuana or drug type) is missing from the list of available species in public Genebanks and only exist as private collections. Most of the Cannabis varieties in the market today are hybrids (700 hybrid strains) with both Cannabis sativa and Cannabis indica genetics. Hence there should be a public Cannabis Genebanks to play a fuller and more effective role in conservation, sustainable use, and exchange of Cannabis genetic resources.

Exogenous application of stress-related signaling molecules affect growth and cannabinoid accumulation in medical cannabis (Cannabis sativa L.).

Medical cannabis (Cannabis sativa L.) is a source of bioactive phytochemicals with promising pharmacological and therapeutic applications. Enhancing the accumulation of valuable bioactive compounds is potentially a way of increasing the economic importance of this crop. Signaling molecules like salicylic acid (SA), jasmonic acid (JA), and γ-aminobutyric acid (GABA) are involved in the regulation of plant development and responses to biotic and abiotic stresses. Moreover, several phytohormones regulate plant trichome formation and elicit the synthesis of secondary metabolites in many plant species in both in vitro and in vivo systems. Therefore, exogenously delivered plant signaling molecules have the potential to modify the chemical profiles of medical cannabis. In this study, we found that the foliar application of SA, methyl jasmonate (MeJA), and GABA produces changes in the accumulation of the two major cannabinoids, cannabidiolic acid (CBDA) and Δ9- tetrahydrocannabinolic acid (THCA), in leaves and inflorescences of a medical cannabis variety. MeJA at 0.1 mM increased the CBDA content in inflorescences by 15.6%, while SA and MeJA at 0.1 mM increased CBDA and THCA accumulation in leaves by up to 57.3%. Treatments did not change the expression of genes participating in the final steps of the biosynthetic pathway of cannabinoids: olivetolic acid cyclase (CsOAC-1 and CsOAC-2), 2-acylphloroglucinol 4-prenyltransferase (CsPT4), cannabidiolic acid synthase (CsCBDAS), and tetrahydrocannabinolic acid synthase (CsTHCAS). Trichome density was not significantly different from the control plants in any treatment. Besides, we found strong correlations between several plant growth parameters and cannabinoid yields, showing a direct link between plant fitness and the production of cannabinoids.

Single-Molecule Real-Time Sequencing of Full-Length Transcriptome and Identification of Genes Related to Male Development in Cannabis sativa.

Female Cannabis sativa plants have important therapeutic properties. The sex ratio of the dioecious cannabis is approximately 1:1. Cultivating homozygous female plants by inducing female plants to produce male flowers is of great practical significance. However, the mechanism underlying cannabis male development remains unclear. In this study, single-molecule real-time (SMRT) sequencing was performed using a mixed sample of female and induced male flowers from the ZYZM1 cannabis variety. A total of 15,241 consensus reads were identified, and 13,657 transcripts were annotated across seven public databases. A total of 48 lncRNAs with an average length of 986.54 bp were identified. In total, 8202 transcripts were annotated as transcription factors, the most common of which were bHLH transcription factors. Moreover, tissue-specific expression pattern analysis showed that 13 MADS transcription factors were highly expressed in male flowers. Furthermore, 232 reads of novel genes were predicted and enriched in lipid metabolism, and qRT-PCR results showed that CER1 may be involved in the development of cannabis male flowers. In addition, 1170 AS events were detected, and two AS events were further validated. Taken together, these results may improve our understanding of the complexity of full-length cannabis transcripts and provide a basis for understanding the molecular mechanism of cannabis male development.