In 2022 and 2024, field surveys were conducted in the Columbia Basin of Washington State, USA, for pathogens transmitted by the beet leafhopper, Circulifer tenellus, including beet curly top virus (BCTV), ‘Candidatus Phytoplasma trifolii’ (CPt), and Spiroplasma citri (S. citri). For these pathogens, an association of symptoms with a causal agent is often complicated by co-infections and the inability to culture CPt (Hu 2021; Rivedal et al. 2022, 2024; Schoener & Wang 2023). In 2022, 35 hemp (Cannabis sativa) plants were observed at the Washington State University (WSU) Research Station in Othello, WA, with typical symptoms of BCTV (a finding published by Jarugula et al. 2023) and we further tested these samples for C. tenellus-associated bacteria. In 2024, 23 and 15 hemp plants observed in Prosser, WA, from the WSU Irrigated Agriculture Research Center and WSU Roza Farm, respectively, with symptoms of CPt infection, including stem fasciation and purpling. Leaf tissue from each plant was ground in liquid nitrogen, DNA extracted using the DNeasy Plant Mini Kit (Qiagen, Inc.), and a multiplex real-time PCR protocol used to detect the C. tenellus-associated pathogens (Swisher Grimm et al. 2023). CPt was identified in 8.6% and 65.8% of plants in 2022 and 2024, respectively, while S. citri was not detected in 2022 but was found in 15.8% of plants in 2024. Co-infections were common. Nested PCR (primers P1/P7 and R16F2n/R16R2, Crosslin et al. 2006) and conventional PCR (primers Trif-SecY-163F, 5ʹ-AGCAGCTAAAAAAGTTAGAAAAAACCTC-3ʹ/Trif-SecY-1040R, 5ʹ-AAATCTAGCGAAAATGATTTTTTGTTTTCA-3ʹ) targeted the CPt 16S rRNA and secY genes, respectively. S. citri infection was confirmed by PCR targeting the spiralin gene (primers SpiralinF/R, Yokomi et al. 2008). All targets were amplified using PrimeSTAR HS DNA polymerase (Takara Bio Inc.). Thermal cycle conditions for CPt 16S rRNA and S. citri spiralin genes consisted of 30 or 40 cycles, respectively, at 98°C for 10 s, 55°C for 15 s, and 72°C for 10 s, followed by a final elongation at 72°C for 5 min. Conditions for CPt secY consisted of 35 cycles at 98°C for 15 s, 55°C for 30 s, and 72°C for 10 s, followed by a final elongation at 72°C for 5 min. Amplicons of size 1,250 bp, 877 bp, and 675 bp for the 16S rRNA, secY, and spiralin genes, respectively, were visualized on an agarose gel stained with GelRed (Sigma-Aldrich) or Ethidium Bromide. For CPt, sequencing of 16S rRNA from seven infected samples (2 from 2022, 5 from 2024) revealed identical sequences (GenBank Accession PV983691) with 100% identity to CPt in U.S. hemp (OQ597521), and sequencing of secY in one sample from 2022 and one from 2024 revealed identical sequences (PX725980), with 100% identity to CPt in U.S. periwinkle (GU004317). For S. citri, two sequenced samples had identical spiralin sequences (PV955037), matching 100% and 99.85% with S. citri found in Oregon cabbage (PV099668) and hemp (OQ969984), respectively. This is the first confirmed report of CPt and S. citri in C. sativa in Washington state. These findings highlight the need to evaluate effects of mollicutes on hemp and underscore the need to develop integrated pest management strategies to reduce vector transmission. In addition, these findings suggest that hemp could serve as a reservoir of the C. tenellus-transmitted pathogens, leading to higher pathogen prevalence across the region, and negatively impacting economically important vegetable and seed crops grown in Washington state that are susceptible to these pathogens.

Cannabidiol (CBD) does not reduce cocaine reward or self-administration in a mouse model of schizophrenia genetic susceptibility.

Cannabidiol (CBD) is a non-psychoactive substance derived from the Cannabis sativa plant, with anti-inflammatory and antioxidant effects. Neuroinflammation is a pathological process triggered by multiple mechanisms in the brain and progresses in a spiral manner, commonly encountered in the pathology of many neurodegenerative diseases. The effects of neuroinflammation on memory problems that begin in the early stages of neurodegenerative diseases such as Alzheimer's disease and Parkinson's disease are discussed. In recent years, the search for effective drugs for these paradoxical pathologies has continued. Thus, our study aimed to evaluate the effect of CBD on learning and memory performance in a mouse model of lipopolysaccharide (LPS)-induced neuroinflammation. An in vivo neuroinflammation model was performed by administering LPS (2mg/kg; ip) to C57BL/6J mice on days 1 and 3. Mice that were given CBD treatment for 21 days were subjected to neurobehavioral tests on the last 4 days. In the open field test, no difference was observed between the groups in terms of spontaneous movements and anxiety. In the novel object recognition test, the movement time to recognise the new object was longer in the CBD treatment group than in the LPS group. Finally, in the passive avoidance test, the delay in moving to the dark compartment was longer in the CBD-treated group than in the LPS group. As a result, the cognitive functions impaired by LPS administration can be improved with CBD treatment, and the study will be carried forward by supporting these behavioural results with neurobiochemical results.

ABSTRACT Brown rot, caused by Phytophthora nicotianae and P. citrophthora , is a major disease of citrus fruit, leading to significant economic losses globally. Conventional fungicides are commonly used to manage this disease, but concerns have been raised due to environmental persistence, human toxicity and the emergence of resistant strains. This study evaluated the spore germination and mycelial growth inhibition of both Phytophthora species by extracts of 31 plant species in vitro and in vivo. DCM/MeOH extracts were tested against both pathogens using a microtiter plate assay and an amended medium assay. All bioactive extracts were further fractionated into seven fractions using solid phase extraction (SPE), which were subsequently tested against both pathogens. Phytochemical profiling was determined by ultra‐performance liquid chromatography quadrupole time‐of‐flight mass spectrometry (UPLC–QTOF‐MS) analysis. Extracts of seven plants ( Artemisia afra , Dombeya rotundifolia , Eucomis regia , Olea europaea , Cannabis sativa , Peltophorum africanum and Mentha longifolia ) inhibited both P. nicotianae and P. citrophthora with a MIC < 1 mg/mL and MFC < 2.5 mg/mL as compared to azoxystrobin (6.0 mg/mL). In the lemon fruit in vivo assays, A. afra reduced brown rot severity by > 50% compared to untreated controls, with superior performance to azoxystrobin, the standard fungicide. UPLC–QTOF‐MS profiling of bio‐active fractions identified scopoletin ( A. afra ), oleuropeinic acid ( O. europaea ) and bergenin ( P. africanum ) as putative contributors to the observed activity. These findings highlight A. afra as a promising natural product and suggest that other extracts warrant further evaluation for use as alternative oomyceticides in citrus post‐harvest protection.

Following a request from the European Commission, the EFSA Panel on Nutrition, Novel Foods and Food Allergens (NDA) was asked to deliver an opinion on a carbon dioxide (CO2) extract from industrial hemp varieties of the plant Cannabis sativa L. as a novel food (NF) pursuant to Regulation (EU) 2015/2283. The NF is proposed to be used in food supplements at maximum use levels ranging from 1.12 mg/day for children above 3 years of age to 3.60 mg/day for adults, excluding pregnant and lactating women. The NF contains cannabidiol (CBD) as well as other cannabinoids. The Panel notes that a substantial proportion of the NF remains uncharacterised. Stability and toxicological tests submitted by the applicant were performed with a material different from the NF under assessment, as it was produced using a different production process (i.e. isopropanol (IPA)-based extraction instead of CO2 supercritical extraction), potentially resulting in different compositional data. Therefore, the absence of adequate stability and of toxicological studies performed with a representative test item prevents the Panel from drawing conclusions on the safety assessment of the NF. Additionally, no data were provided by the applicant to address the safety of the NF in the general population above 3 years of age. Therefore, the safety of the NF under the proposed conditions of use cannot be established.

Preclinical Study of Cannflavins A and B Action Against Glioblastoma Cells

Modulatory function of cannabidiol on the extinction and reinstatement of methamphetamine-seeking behavior through the D2-like dopamine receptors in the dentate gyrus.

Cannflavin B ameliorates behavioural and neuronal systems alterations in adolescent rats exposed to prenatal valproic acid.

Background/AimMesenchymal stem cells (MSCs) are used to treat various degenerative diseases. However, their therapeutic potential is limited by cellular aging during in vitro cultivation. This study aimed to explore whether cannabidiol (CBD) can delay MSC aging by enhancing the expression of Sirtuin 1 (SIRT1) and autophagy, two key anti-aging regulators.Materials and MethodsCBD, the most important non-psychotomimetic phytocannabinoid derived from the Cannabis sativa plant, was used to up-regulate SIRT1 and autophagy in order to maintain MSC stemness. MSCs were treated with CBD and assessed for cell viability, doubling time, key gene/protein expression, relative senescence-associated β-galactosidase (SA-β-gal) assay, relative telomere length, and telomerase expression.ResultsCBD significantly increased the expression of SIRT1 and autophagy-related markers in MSCs. Furthermore, CBD preserved MSC stemness by promoting the deacetylation of SRY-box transcription factor 2 (SOX2) through SIRT1, and delayed cellular senescence by enhancing autophagy, reducing SA-β-gal activity, maintaining proliferation capacity, and supporting telomere function.ConclusionCBD promotes MSC stemness and delays cellular senescence, potentially through the activation of SIRT1 and autophagy. These findings suggest that CBD may serve as a promising agent for preserving MSC function in regenerative medicine.

Cannabinoids, such as tetrahydrocannabinolic acid (THCA), cannabidiolic acid (CBDA) and cannabichromenic acid (CBCA), are bioactive and medicinally relevant compounds found in the cannabis plant (Cannabis sativa L.). These three compounds are synthesised from a single precursor, cannabigerolic acid (CBGA), through regioselective reactions catalysed by different cannabinoid oxidocyclase enzymes. Despite the importance of cannabinoid oxidocyclases for determining cannabis chemotype and properties, the functional evolution and molecular mechanism of this enzyme family remain poorly understood. To address this gap, we combined ancestral sequence reconstruction and heterologous expression to resurrect and functionally characterise three ancestral cannabinoid oxidocyclases. Results showed that the ability to metabolise CBGA originated in a recent ancestor of cannabis and that early cannabinoid oxidocyclases were promiscuous enzymes producing all three THCA, CBDA and CBCA. Gene duplication and diversification later facilitated enzyme subfunctionalisation, leading to extant, highly-specialised THCA and CBDA synthases. Through rational engineering of these ancestors, we designed hybrid enzymes which allowed identifying key amino acid mutations underlying the functional evolution of cannabinoid oxidocyclases. Ancestral and hybrid enzymes also displayed unique activities and proved to be easier to produce heterologously than their extant counterparts. Overall, this study contributes to understanding the origin, evolution and molecular mechanism of cannabinoid oxidocyclases, which opens new perspectives for breeding, biotechnological and medicinal applications.

It is important to understand how water stress treatments (water shortage and waterlogging) affect plant physiology and cannabinoid levels in cannabis plants. This study evaluated the effects of different periods (7, 14, and 21 days) of water stress (water shortage vs. waterlogging) on growth, leaf physiology, and cannabinoid accumulation in Cannabis sativa plants at vegetative and reproductive stages. The first experiment evaluated the effects of 7 days of water stress treatments during the vegetative stage. The second study examined the physiological responses to different durations (7, 14, and 21 days) of water stress treatments during the reproductive phase. Waterlogging for seven days during the vegetative stage had a more negative impact on growth parameters, especially root length (17.2 cm), compared to water shortage (31.6 cm) and control plants (17.2 cm). During the reproductive stage, water shortage for 21 days significantly reduced stomatal conductance (gs) (34.9 µmol·m-2·s-1), leaf area index (LAI) (1.01), total plant dry weight (TPDW) (52 g), and photosynthetic efficiency (Phi2:0.2) compared to waterlogging (gs: 113 µmol·m-2·s-1; LAI: 2.39; TPDW: 62 g; Phi:0.3), and control plants (gs: 226 µmol·m-2·s-1; LAI: 3.82; TPDW: 80 g; Phi:0.65). Both water stress conditions decreased cannabinoid content when the stress period was prolonged during the reproductive stage (Total CBD: 45 and 41 mg·g-1 for water shortage and waterlogging, respectively) compared with control plants (Total CBD: 105 mg·g-1). This study found that water stress conditions, both waterlogging and water shortage, influenced cannabinoid biosynthesis during the reproductive stage.

Abstract Cannabis revenue has risen in the United States (US) from $16.5B in 2018 to $45.4B in 2025, which can be attributed to the legalization of hemp in 2018 with the passage of the Agricultural Improvement Act of 2018 (2018 Farm Bill) and the surge of Cannabis -derived finished products. Hemp was defined as any Cannabis sativa plant having a Δ 9 -tetrahydrocannabinol (Δ 9 -THC) mass fraction of ≤ 0.3% on a dry weight basis. The legalization of finished products with higher THC levels has continued to increase. As of April 2025, medical marijuana is legal in 41 states, and recreational marijuana is legal in 24 states (and the District of Columbia). After the passage of the 2018 Farm Bill, there was a significant increase in hemp grown in the US, resulting in the supply outweighing the user demand. As a result, scientists developed a process for the synthesis of Δ 8 -tetrahydrocannabinol (Δ 8 -THC), Δ 10 -tetrahydrocannabinol (Δ 10 -THC), and other cannabinoids from hemp extracts containing high amounts of cannabidiol (CBD). Δ 8 -THC and Δ 10 -THC are popular synthetic cannabinoids in Cannabis -derived finished products because they exhibit similar psychoactive effects to Δ 9 -THC. However, these synthetic processes usually produce a combination of isomeric cannabinoids. One of the more common forms of these finished products is called Cannabis concentrates, which include but are not limited to oils, vapes, waxes, distillates, and isolates. The Chemical Sciences Division (CSD) at the National Institute of Standards and Technology (NIST) previously developed an analytical method for the determination of CBD, Δ 9 -THC, and other cannabinoids in one form of concentrates, oils, by liquid chromatography (LC) with a photodiode array (PDA) detector for ultraviolet (UV) absorbance detection. Hemp oils are diluted with methanol and shaken for 15 min using routine laboratory equipment. This manuscript expands the LC-PDA method to include Δ 8 -THC-based and CBD-based concentrates to re-evaluate its performance for an expanded range of Cannabis samples. The results presented here highlight that other cannabinoids are often detected in these samples, leading to chromatographic issues for the accurate measurement of Δ 9 -THC. NIST CSD collaborated with Kaycha Labs in 2024 to better understand the concentrates currently available in the Cannabis industry and provide an in-depth study of 23,000 concentrate samples to understand the prevalence of these types of samples.

Recent innovations in cannabinoid chemistry, biology, and biosynthesis.

Background/Objectives: Cannabidiol (CBD) is a non-psychoactive compound found in the Cannabis sativa plant. Due to its broad therapeutic potential, CBD is often incorporated into various pharmaceutical formulations. This study aimed to evaluate homogenous (oil-based) and heterogeneous (emulsion-based) liquid preparations of CBD using different fatty oils and provide a comprehensive comparative framework for the development of stable liquid dosage forms of cannabidiol (CBD), with direct applications in pharmaceutical formulations. Methods: The oils and emulsions were qualitatively analysed to assess their stability and suitability as CBD carriers. Ultraviolet (UV) spectrophotometry and High-Performance Liquid Chromatography (HPLC) were employed for quantifying CBD in the formulations and also characterising them in terms of product quality. Results: The results indicated that sunflower oil is the most stable and analytically compatible matrix, with CBD recovery close to 100% and minimal degradation over time. Conversely, linseed and pumpkin seed oils exhibited significant analytical interference and oxidative instability. Oil-in-water emulsions prepared with a 4% Tween 80/Span 80 mixture demonstrated optimal physical stability and droplet size distribution. Conclusions: Overall, both formulations can be regarded as suitable pharmaceutical carriers for CBD delivery.

Cannabidiol (CBD), the second most abundant of the active compounds found in the Cannabis sativa plant, is nonaddictive and of increasing interest due to its potential therapeutic utilities. The actions of CBD are mediated by cannabinoid receptors and other targets, both G protein-coupled receptors (GPCRs) and non-GPCR proteins. It has recently been shown that CBD is not an orthosteric ligand of the adenosine A2A receptor (A2AR), but rather a negative allosteric modulator. Because of the multitude non-conserved allosteric pockets in GPCRs, the binding mode of CBD to A2AR remains unknown. To fill this knowledge gap, and due to the therapeutic relevance of CBD and A2AR, we have used metadynamics simulations and site-directed mutagenesis to identify the binding mode of CBD. These theoretical and experimental results show that the allosteric binding site of CBD is along the ligand binding pathway of adenosine to A2AR, near the orthosteric binding site.

Apical stem cuttings are the primary method for propagation in medicinal cannabis, yet propagation has not been studied as extensively as the later stages of crop cultivation. This study examined how mother plant age and photosynthetic photon flux density (PPFD) during mother plant cultivation and propagation affect rooting, growth, and development of apical stem cuttings. Mother plants ( Cannabis sativa ‘Original Blitz’ and ‘King Harmony’) were grown in climate-controlled chambers under two light intensities (400 and 800 μmol·m −2 ·s −1 ) for up to 6 months. Apical stem cuttings were excised every 3 weeks and subsequently propagated without externally applied auxin for 3 weeks under three light intensities (50, 150, and 250 μmol·m −2 ·s −1 ). Mother plant age did not affect rooting (root dry mass and fraction of rooted cuttings). However, older mother plants exhibited decreased cutting dry mass at severance, which coincided with a reduced leaf area. The light intensity during mother plant cultivation had genotype-specific effects, with rooting either reduced or unaffected for the higher light intensity. This reduction coincided with an accumulation of starch and soluble sugar at the stem base at severance, while auxin concentrations in the apex, leaf, and stem base were unaffected by light intensity during mother plant cultivation. In contrast, light intensity during propagation did not affect the fraction of rooted cuttings. However, higher light intensity increased root and cutting dry mass. These findings indicate that mother plant age, up to 6 months, does not impact rooting in stem cuttings. However, higher light intensity during mother plant cultivation reduced rooting genotype-dependently, whereas higher light intensity during propagation increased root dry mass without affecting fraction of rooted cuttings.

Unlocking the resorption potential of cannabidiolic acid: A comprehensive in vitro and in vivo bioavailability study.

Cannabidiol (CBD), a non-psychoactive compound sourced from the Cannabis sativa plant, has gained significant interest in the skincare industry for its therapeutic properties. Review highlights anti-inflammatory and antioxidant properties of Cannabidiol, focusing on its role in supporting skin health. Inflammation and oxidative stress are major factors in skin issues such as acne, eczema, psoriasis, and premature aging. By interacting with the endocannabinoid system, CBD helps regulate inflammatory processes, potentially alleviating these conditions. Additionally, its potent antioxidant activity aids in neutralizing free radicals, shielding the skin from environmental aggressors like UV rays and pollution. Recent studies emphasize CBD's capacity to reduce pro-inflammatory cytokine production and combat oxidative stress, showcasing its potential in preventing and managing various skin conditions. Furthermore, its calming effects make it an ideal choice for sensitive skin, offering a natural alternative to conventional skincare ingredients. In summary, CBD shows great promise as a multifunctional ingredient in skincare, delivering both anti-inflammatory and antioxidant benefits. As interest in natural, effective skincare solutions continues to grow, further studies are needed to explore CBD's mechanisms and evaluate its long-term impact in dermatology.

IntroductionResearch on optimizing light intensity to maximize phytochemical production during hemp flowering is limited, despite growing global demand. We investigated the effects of light-emitting diode (LED) intensity on hemp growth, cannabinoid content, and gene expression.MethodsHemp plants (Cannabis sativa 'Queen Dream') were grown under white LEDs at light intensities of 200, 400, and 600 μmol·m−2·s−1 with a 12/12 h photoperiod for 35 days during the flowering stage.ResultsThe dry mass of stems, leaves, and flowers increased linearly with increasing light intensity. Cannabinoid analysis revealed that levels of cannabidiol (CBD), cannabidiolic acid, and tetrahydrocannabinolic acid increased linearly with light intensity, reaching the highest levels at 600 μmol·m−2·s−1. Total CBD increased by 36.88% at 600 μmol·m−2·s−1 compared to 200 μmol·m−2·s−1. CBD yield per plant also increased linearly across the entire light intensity range. Gene expression analysis revealed a coordinated upregulation of genes involved in the hexanoate–olivetolic acid–cannabigerolic acid–cannabinoid biosynthesis pathway under high light intensity, with a notable increase in cannabidiolic acid synthase (CBDAS) expression.ConclusionThese findings demonstrate that a light intensity of 600 μmol·m−2·s−1 effectively enhances both biomass and cannabinoid accumulation at the flowering stage, providing valuable insights for controlled-environment hemp cultivation aimed at maximizing CBD yield.

BackgroundPlum is a nutritionally and medicinally valuable fruit with high economic value; however, in temperate regions, its yield continuously suffers from bacterial canker caused by Pseudomonas syringae pathovars.ResultsTo bridge the research gap, a study was conducted to optimize the use of rhizobacteria and biogenic silver nanoparticles (AgNPs) for controlling bacterial canker in plums. Rhizobacteria were isolated from plum orchards and identified (through 16 S rRNA gene sequencing, phylogenetic analysis using MEGA) and further evaluated for plant growth-promoting characteristics (siderophore production, production of hydrogen cyanide, solubilization of phosphate, and biofilm formation). The results revealed that 7 isolates out of 19 rhizobacterial isolates produced biofilm, of which Rw-1, Rw-6, Rw-7, and Hj-2 were Pseudomonas fluorescens, while Rw-12 and Hj-5 were identified as Bacillus subtilis. Besides, biogenic AgNPs were prepared in different concentrations (0.2%, 0.4%, and 0.6%) by using Cannabis sativa plant extract. X-ray diffraction (indicated the size of crystalline green silver nanoparticles of ~ 20 nm), Fourier-transform infrared spectroscopy (confirmed the presence of alkyne molecules, polyphenols, and carbonyl compounds on the AgNPs surface), and scanning electron microscopy (SEM) were used for characterization of the nanoparticles. Additionally, the antibacterial activity of AgNPs was determined by the disk diffusion assay with 18-hour cultures of P. syringae. In vitro experiments showed that the highest inhibition zone (24.32 mm) was caused by the 0.6% AgNPs concentration, proving the strongest antibacterial activity. The subsequent glasshouse tests confirmed that a consortium of two strains of Bacillus with 0.6% AgNPs allowed the lowest disease incidence (18.13%), proving efficient disease suppression. Combination of Bacillus and Pseudomonas strains with 0.6% AgNPs recorded significantly higher disease incidence, with the probable reason being incompatibility among these bacterial genera.ConclusionsThe research findings emphasize the potential of applying compatible rhizobacterial consortia and biogenic (C. sativa) silver nanoparticles as a potent substitute for synthetic pesticides in controlling plum bacterial canker.