Search Results (47)

Biostimulants represent a sustainable strategy to enhance the therapeutic potential of medicinal plants, which often exhibit low and variable levels of bioactive compounds. Cannabis sativa, a medicinally important species, produces diverse cannabinoids, such as THC, CBD, CBG, and CBC, whose profiles depend on plant chemotype and determine pharmacological activity. We developed a novel plant-based biostimulant, Tricostimulant™, to optimize cannabinoid production in Cannabis sativa. Field trials demonstrated increased biomass and selective enhancement of cannabinoid content. In high-CBD chemotypes, Tricostimulant™ was associated with higher CBD and CBG without relevant changes in THC levels, whereas in high-THC chemotypes, higher THC values were observed without evident variation in CBD. The most pronounced differences were observed when the biostimulant was applied during the vegetative stage, highlighting the importance of application timing. These results indicate the potential of Tricostimulant™ to modulate cannabinoid profiles, contributing to improved optimization and standardization of cannabis-based therapeutics. Further research is required to confirm these findings and elucidate the underlying mechanisms of biostimulant action. Full article

The bioproduction of high-value molecules offers a sustainable and cost-effective alternative to traditional extraction and chemical synthesis, particularly for complex metabolites like cannabinoids (CBs), which have therapeutic potential for neurodegenerative diseases. The marine diatom Phaeodactylum tricornutum presents a promising chassis for CB biosynthesis due to its high lipid content, essential building blocks to biosynthesize CBs. In this study, we explored the feasibility of producing olivetolic acid (OA), the key CB precursor, using a hybrid-type polyketide synthase, SteelyA, from Dictyostelium discoideum. Unlike the native Cannabis sativa enzymes—tetraketide synthase and olivetolic acid cyclase—which exhibit low productivity and stability in diatoms, SteelyA was expected to offer an alternative biosynthetic route. Heterologous production in P. tricornutum resulted in a C-terminal fragment of the SteelyA enzyme, suggesting partial expression or processing of the very high-molecular-weight (352 kDa) SteelyA protein over six months without affecting cellular growth. However, HPLC-MS analysis did not detect intracellular OA or its derivatives in vivo and in vitro, suggesting enzymatic inactivity or metabolic limitations. These negative findings highlight the need for further investigation into the metabolic and proteomic requirements for CB precursor biosynthesis in diatoms, guiding future optimization strategies for sustainable cannabinoid production. Full article

Hemp (Cannabis sativa L.) is a polymorphic species that synthesizes an array of bioactive metabolites, with cannabinoids and terpenoids constituting the major chemical classes. Until recently, the lack of legislative framework led to limited research on hemp’s plant protection and nutrition. Biostimulants have recently attracted scientific attention as sustainable alternatives to plant protection products (PPPs). Herein, we investigated the effects of biostimulant harpin (αβ) proteins and the PPP polysaccharide laminarin on hemp (cv. Futura 75), employing GC/EI/MS and ¹H NMR metabolomics. Analyses demonstrated that treatments induced distinct shifts in the metabolism of the plants, thus, enabling the discovery of metabolite-biomarkers of physiological adaptation, defense mechanisms (α-linolenic acid), and bioactivity (cannabinoids). Harpin and laminarin altered the concentration of bioactive compounds such as cannabidiol, essential amino acids including L-phenylalanine and GABA, salicylate, and caffeate. Pathway analysis revealed treatment-specific modulation of key metabolic networks, with harpin triggering early, yet transient activation of phenylpropanoid- and amino acid-related pathways before broad repression, whereas laminarin maintained a more balanced regulation, sustaining defense-related biosynthesis while preserving core primary metabolism. Results advance the understanding of molecular mechanisms underlying biostimulants’ action in hemp and support their potential for improving plant health and attributes of cannabis-derived products, providing insights for its sustainable cultivation. Full article

Trichomes in cannabis (Cannabis sativa L.) are specialized structures responsible for cannabinoid and terpene biosynthesis, making their density a critical parameter for both research and industrial applications. However, consistent trichome density assessment remains challenging due to anatomical variability and the absence of standardized methodologies. This review critically examines the existing literature on trichome quantification across key floral structures—such as bracts, sugar leaves, calyxes, and the main cola—to identify the most reliable sites and practices for accurate evaluation. Evidence suggests that bracts represent the most consistent sampling unit, given their homogeneous trichome distribution and elevated cannabinoid concentration. Whilst sugar leaves and calyxes are also frequently analyzed, their morphological variability requires cautious interpretation. Furthermore, trichome shape, size, maturity, and vegetal surface expansion/shrinkage during stress must be considered when correlating density with secondary metabolite production. We also highlight the advantages of using more than only one floral structure and integrating microscopic imaging and software-assisted analysis to enhance reproducibility and accuracy. By synthesizing current methodologies and proposing pathways for standardization, this review aims to support more robust trichome assessment protocols, ultimately improving cannabinoid yield optimization, quality control, broader cannabis research frameworks, and an important aesthetic parameter for consumers. Future research efforts should focus on advancing imaging methodologies and optimizing sampling protocols to further improve the precision and reproducibility of trichome density and cannabinoid analyses. Full article

Cannabis sativa produces pharmacologically valuable cannabinoids. In this study, we developed and optimized a transient transformation system using Cannabis sativa ‘Cheungsam’ to facilitate gene functional analysis. Various experimental conditions, including plant developmental stages, light conditions, Agrobacterium strains, tissue types, and physical treatments such as sonication and vacuum infiltration, were systematically evaluated using GUS histochemical staining and qPCR analysis. Among these, 7-day-old seedlings cultured under dark conditions and transformed with the GV3101 strain exhibited high transformation efficiency. Leaf tissue showed a higher GUS staining proportion and GUS staining area compared to hypocotyl and cotyledon tissues. The application of a combination of sonication and vacuum infiltration techniques resulted in the most intense GUS expression. Using the optimized protocol, we introduced a recombinant vector carrying CsCBDAS2, a key gene in cannabidiol (CBD) biosynthesis. qPCR analysis revealed that CsCBDAS2 overexpression led to significant upregulation of multiple upstream CBD biosynthetic genes (CsOAC, CsGOT, CsPT1, CsPT4, CsCBDAS1, and CsCBDAS2) and the transcription factor (TF) CsWRKY20, suggesting coordinated co-expression and potential involvement of a transcriptional feedback loop. These results demonstrate the effectiveness of our transient transformation system and provide insights into the regulatory mechanisms of cannabinoid biosynthesis in cannabis. Full article

Water availability is a key factor affecting both morphological development and secondary metabolite production in Cannabis sativa L. This study evaluated the effects of water stress applied during the vegetative and flowering stages on plant performance, cannabinoid concentration, and terpene composition in two Chemotype III (cannabidiol-dominant) varieties. Plants were subjected to moderate and severe water stress, and responses were assessed through biomass measurements, GC-MS analyses, and multivariate statistics. Water stress significantly influenced biomass allocation, with increased dry biomass but reduced harvest index, particularly under flowering-stage stress. Cannabidiol (CBD) content declined with increasing stress, while tetrahydrocannabinol (THC) levels increased under vegetative stress, indicating a stress-induced shift in cannabinoid biosynthesis. Cannabinol (CBN) levels also increased, suggesting enhanced THC degradation. Terpene composition was predominantly genotype-driven. PCA-MANOVA showed significant effects of variety, stress level, and their interaction, yet only minor volatiles were modulated by stress, while the most abundant terpenes remained stable across treatments, preserving the varietal aroma profile. These results underline the importance of genetic background and irrigation timing in determining cannabis yield and quality. Optimized water management is essential to ensure phytochemical consistency and sustainable production, especially in high-value medicinal and aromatic applications. Full article

The endocannabinoid system is a complex communication system involved in maintaining homeostasis in various physiological processes, including metabolism, immune response, pain modulation, and neuroprotection. Endocannabinoids, mainly anandamide and 2-arachidonoylglycerol, are natural ligands of the cannabinoid receptors CB1 and CB2, which are widely distributed throughout the central nervous system and peripheral tissues. Their biosynthesis, degradation, and interaction with other signaling pathways play crucial roles in both health and disease. This article provides a comprehensive overview of the physiological and pathological roles of endocannabinoids, discusses their potential as therapeutic targets, and highlights recent advances in endocannabinoid-based treatments. Full article

The diverse hormonal treatments applied to hemp (Cannabis sativa L.) carry significant implications for cultivation, and yield optimization across a range of applications, including fiber, seed, oil production, and the enhancement of medicinal compounds. However, there is no evidence concerning the long-term consequences of hormonal treatment. To determine the connection between the effects of hormonal treatment and cannabinoid accumulation, hemp plants were treated with γ-aminobutyric acid (GABA), abscisic acid (ABA), and salicylic acid (SA) to investigate their effects on gene expression and cannabinoid content levels in female inflorescences at 3 days and 4 weeks after treatment. The treatments influenced the transcript levels of five key cannabinoid biosynthesis genes, with 1.0 mM GABA significantly increasing OAC, THCAS, and CBCAS transcripts within 48 to 72 h. Additionally, 1.0 mM GABA led to a significant increase in tetrahydrocannabinol content by day three and significant increases in total cannabidiol and cannabinoid by week four. In addition, both ABA and SA induced transient, dose-dependent increases or decreases in gene expressions, but cannabinoid accumulation at 4 weeks showed no significant changes compared to the control. These results provide valuable insights for hormonal application in cultivation and the development of traits that enhance cannabinoid production in cannabis cultivation, which could significantly contribute to optimizing industrial applications. Full article

The endocannabinoid system (ECS) plays a crucial role in maintaining homeostasis by regulating immune response, energy metabolism, cognitive functions, and neuronal activity. It consists of endocannabinoids (eCBs), cannabinoid receptors (CBRs), and enzymes involved in eCB biosynthesis and degradation. Increasing evidence highlights the involvement of the ECS under several pathological conditions, making it a promising therapeutic target. Recent research efforts have focused on modulating endogenous eCB levels, particularly through the inhibition of fatty acid amide hydrolase (FAAH), the main catabolic enzyme of the major eCB anandamide. Natural substances, including plant extracts and purified compounds, can inhibit FAAH and represent a promising area of pharmacological research. Natural FAAH inhibitors are particularly attractive due to their potentially lower toxicity compared to synthetic compounds, making them safer candidates for therapeutic applications. Phytocannabinoids, flavonoids, and flavolignans have been shown to efficiently inhibit FAAH. The structural diversity and bioactivity of these natural substances provide a valuable alternative to synthetic inhibitors, and may open new avenues for developing innovative pharmacological tools. Full article

The rising global demand for medicinal cannabis necessitates the optimization of cultivation, harvesting, and extraction techniques to maximize cannabinoid yield and purity. This study investigates the Foi Thong Phu Pha Yon strain under controlled environmental conditions, evaluating the effects of temperature, humidity, CO₂ concentration, and light exposure on plant growth and cannabinoid biosynthesis. A total of 170 seeds were germinated, with an 85% germination success rate, and various growth strategies, including soil composition, nutrient application, and irrigation methods, were tested to determine the most effective approach. The research findings indicate that vegetative growth was optimal at 27 °C, 70% humidity, and 1200 ppm CO₂ while flowering required a reduced temperature (22 °C), lower humidity (50%), and elevated CO₂ levels (1900 ppm) to enhance cannabinoid production and prevent disease. Furthermore, harvest timing significantly influenced CBD yield, with peak cannabinoid content observed when 80% of trichomes were cloudy white. Over two growing cycles, this study produced 43,200 g of fresh buds, resulting in 7560 g of dried cannabis buds. The extraction process, utilizing dynamic maceration with 95% ethanol, followed by winterization and chromatography, yielded 2343.60 g of cannabis extract, including 589.68 g of CBD, with an average purity of 86.599%. Advanced techniques such as flash chromatography and distillation further refined the CBD isolate, ensuring pharmaceutical-grade quality. These findings highlight the effectiveness of precise environmental control, strategic harvesting, and advanced extraction methodologies in optimizing cannabis production. This research provides valuable insights for agricultural researchers, policymakers, and the pharmaceutical industry, supporting sustainable cultivation practices and improved product quality in the expanding medicinal cannabis market. Full article

Cannabidiol (CBD), a non-psychoactive cannabinoid, has attracted significant research interest due to its antioxidant, anti-inflammatory, and neuroprotective properties. As a versatile scaffold in drug discovery, CBD has been widely explored for developing novel therapeutics. In this study, we synthesized and evaluated the anti-tyrosinase activity of CBD-based thiosemicarbazones. Structure–activity relationship (SAR) analyses were conducted to assess the impact of various functional groups on tyrosinase inhibition, including an evaluation of inhibitory kinetics for selected compounds. The synthesized derivatives demonstrated potent tyrosinase inhibition, with activity comparable to kojic acid, a standard tyrosinase inhibitor. Given the crucial role of tyrosinase in melanin biosynthesis, these findings suggest that CBD-based thiosemicarbazones could serve as promising candidates for managing tyrosinase-related disorders, including hyperpigmentation and melanogenesis-related conditions. Moreover, the presence of thiosemicarbazone moieties may contribute to the observed inhibitory effects, potentially through metal chelation at the enzyme’s active site. This study provides valuable insights into the design of CBD-derived inhibitors targeting tyrosinase. Further optimization and in-depth biological evaluation are warranted to explore their full therapeutic potential. Full article

The endocannabinoid system (ECS), composed of receptors, endocannabinoids, and enzymes that regulate biosynthesis and degradation, plays a fundamental role in the physiology and pathology of the gastrointestinal tract, particularly in the small and large intestine and liver. Specifically, cannabinoid receptor type 1 (CB1R) and cannabinoid receptor type 2 (CB2R), located principally in the nervous system and immune cells, orchestrate processes such as intestinal motility, intestinal and hepatic inflammation, and energy metabolism, respectively. The main endocannabinoids, anandamide (AEA) and 2-arachidonoylglycerol (2-AG), influence appetite, body weight regulation, and inflammatory states and thus have implications in obesity, non-alcoholic fatty liver disease (NAFLD) and irritable bowel syndrome (IBS). Recent studies have highlighted the therapeutic potential of targeting the ECS to modulate gastrointestinal and metabolic diseases. In particular, peripheral CB1R antagonists and CB2R agonists have shown efficacy in treating intestinal inflammation, reducing hepatic steatosis, and controlling IBS symptoms. Moreover, the ECS is emerging as a potential target for the treatment of colorectal cancer, acting on cell proliferation and apoptosis. This review highlights the opportunity to exploit the endocannabinoid system in the search for innovative therapeutic strategies, emphasizing the importance of a targeted approach to optimize treatment efficacy and minimize side effects. Full article

Both hydrogen sulfide and endocannabinoids can protect the neural retina from toxic insults under in vitro and in vivo conditions. Purpose: The aim of the present study was two-fold: (a) to examine the neuroprotective action of cannabinoids [methanandamide and 2-arachidonyl glycerol (2-AG)] against hydrogen peroxide (H₂O₂)-induced oxidative damage in the isolated bovine retina and (b) to evaluate the role of endogenously biosynthesized hydrogen sulfide (H₂S) in the inhibitory actions of cannabinoids on the oxidative stress in the bovine retina. Methods: Isolated neural retinas from cows were exposed to oxidative damage using H₂O₂ (100 µM) for 10 min. When used, tissues were pretreated with methanandamide (1 nM–100 nM) and 2-AG (1–10 µM) for 30 min before a 10 min treatment with H₂O₂ (100 µM). In some experiments, retinas were pretreated with inhibitors of the biosynthesis of H₂S [cystathionine β-synthase/cystathionine γ-lyase (CBS/CSE), aminooxyacetic acid, AOAA 30 µM, or 3-mercaptopyruvate sulfurtransferase (3MST), α-keto-butyric acid, KBA 1 mM] and the CB1-receptor antagonist, AM251 (100 nM) for 30 min before treatment with methanandamide (1 nM–100 µM). Enzyme immunoassay measurement of 8-epi PGF2α (8-isoprostane) levels was performed to assess lipid peroxidation in retinal tissues. Results: In the presence of H₂O₂ (100 µM), methanandamide (1 nM–100 µM) and 2-AG (1–10 µM) significantly (p < 0.001) blocked the H₂O₂-induced elevation in 8-isoprostane levels in the isolated bovine retina. In the presence of the CB1 antagonist AM251 (100 nM), the effect of methanandamide (1 nM) on the H₂O₂-induced 8-isoprostane production was significantly (p < 0.001) attenuated. While AOAA (30 µM) had no significant (p > 0.05) effect on the inhibition of H₂O₂-induced oxidative stress elicited by methanandamide, KBA (1 mM) reversed the neuroprotective action of methanandamide. Conclusions: The cannabinoids, methanandamide and 2-AG can prevent H₂O₂-induced oxidative stress in the isolated bovine retina. The neuroprotective actions of cannabinoids are partially dependent upon the activation of the CB1 receptors and endogenous production of H₂S via the 3-MST/CAT pathway. Full article

Cannabis sativa (C. sativa L.) has garnered significant attention worldwide due to its widespread use as a pharmaceutical agent. With the increasing clinical application of C. sativa and cannabinoid therapeutics, there is strong interest in the development of superior plant varieties and optimisation of growth conditions to enhance secondary metabolite yield. Our RNA sequencing analysis revealed differential expression of hormone-related transcripts in developing C. sativa trichomes, suggesting the involvement of hormone signalling pathways in cannabinoid production. Leveraging the potency of exogenous hormones on plants, this study sought to determine if the application of cytokinin (CK), gibberellic acid (GA) and jasmonic acid (JA) modified trichome morphology and the cannabinoid profile over an 8-week period following the induction of flowering. Exogenous hormone application led to alterations in trichome morphology, with each treatment significantly reducing trichome head width by the final week of assessment. Interestingly, GA application also resulted in a significant reduction in the concentration of Δ-9-tetrahydrocannabinol (THC), Δ-9-tetrahydrocannabinolic acid (THCA), cannabidiol (CBD) and cannabidiolic acid (CBDA) by week 8 post floral induction, however, JA and CK treatment did not consistently modulate the accumulation of these cannabinoids. The minor cannabinoids, cannabidivaranic acid (CBDVA), cannabicyclolic acid (CBLA), cannabicyclol (CBL), cannabichromene (CBC), cannabigerolic acid (CBGA) and cannabigerol (CBG), were also affected by hormone treatments, with varying degrees of accumulation observed. These findings underscore the intricate interplay between phytohormones and secondary metabolite biosynthesis in C. sativa. Our study highlights the potential of hormone modulation as a strategy to enhance cannabinoid yield and offers some insights into the regulatory mechanisms governing cannabinoid biosynthesis in C. sativa trichomes. Full article

A cutaneous mast cell tumor (cMCT) is among the most common tumors in dogs. Endocannabinoids (eCBs) belong to the endocannabinoid system (ECS), which involves also cannabinoid receptors and an enzymatic system of biosynthesis and degradation. In this study, plasma levels of N-arachidonoylethanolamine (AEA), 2-arachidonoylglycerol (2-AG), N-palmitoylethanolamine (PEA), and N-oleoylethanolamine (OEA) were evaluated in 17 dogs with MCTs of varying histological grades and clinical stages, as well as in a control group of 11 dogs. Dogs affected by cMCT had higher plasma levels of 2-AG (p = 0.0001) and lower levels of AEA (p = 0.0012) and PEA (p = 0.0075) compared to the control group, while no differences were observed at the OEA level between healthy and cMCT dogs (p = 0.9264). The ability of eCBs to help discriminate between healthy and cMCT dogs was interrogated through the area under the ROC curve (AUC). An accuracy of 0.98 (95% confidence interval [CI], 0.94–1.02) was found for 2-AG, of 0.85 (95% CI, 0.71–0.99) for AEA, and of 0.81% for PEA (95% CI, 0.64–0.69). Values > 52.75 pmol/mL for 2-AG showed 94% sensitivity and 90% specificity in distinguishing cMCT. This is the first study to demonstrate alterations in plasmatic levels of eCBs in dogs affected by MCTs, suggesting the significance of these biomarkers in the tumorigenic process and their potential use as biomarkers in the future. Full article