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(hemp) is a source of various biologically active compounds, for instance, cannabinoids, terpenes and phenolic compounds, which exhibit antibacterial, antifungal, anti-inflammatory and anticancer properties. With the purpose of expanding the auxiliary application of in the field of bio-nanotechnology, we explored the plant for green and efficient synthesis of gold nanoparticles (AuNPs) and silver nanoparticles (AgNPs). The nanoparticles were synthesized by utilizing an aqueous extract of stem separated into two different fractions (cortex and core [xylem part]) without any additional reducing, stabilizing and capping agents. In the synthesis of AuNPs using the cortex enriched in bast fibers, fiber-AuNPs (F-AuNPs) were achieved. When using the core part of the stem, which is enriched with phenolic compounds such as alkaloids and cannabinoids, core-AuNPs (C-AuNPs) and core-AgNPs (C-AgNPs) were formed. Synthesized nanoparticles were character-ized by UV-visible analysis, transmission electron microscopy, atomic force microscopy, dynamic light scattering, Fourier transform infrared, and matrix-assisted laser desorption/ionization time-of-flight. In addition, the stable nature of nanoparticles has been shown by thermogravimetric analysis and inductively coupled plasma mass spectrometry (ICP-MS). Finally, the AgNPs were explored for the inhibition of and biofilms. The synthesized nanoparticles were crystalline with an average diameter between 12 and 18 nm for F-AuNPs and C-AuNPs and in the range of 20-40 nm for C-AgNPs. ICP-MS analysis revealed concentrations of synthesized nanoparticles as 0.7, 4.5 and 3.6 mg/mL for F-AuNPs, C-AuNPs and C-AgNPs, respectively. Fourier transform infrared spectroscopy revealed the presence of flavonoids, cannabinoids, terpenes and phenols on the nanoparticle surface, which could be responsible for reducing the salts to nanoparticles and further stabilizing them. In addition, the stable nature of synthesized nanoparticles has been shown by thermogravimetric analysis and ICP-MS. Finally, the AgNPs were explored for the inhibition of and biofilms. The nanoparticles exhibited minimum inhibitory concentration values of 6.25 and 5 µg/mL and minimum bactericidal concentration values of 12.5 and 25 µg/mL against and , respectively.

For more than a century, Cannabis was considered a narcotic and has been banned by lawmakers all over the world. In recent years, interest in this plant has increased due to its therapeutic potential, in addition to a very interesting chemical composition, characterized by the presence of an atypical family of molecules known as phytocannabinoids. With this emerging interest, it is very important to take stock of what research has been conducted so far on the chemistry and biology of Cannabis sativa. The aim of this review is to describe the traditional uses, chemical composition and biological activities of different parts of this plant, as well as the molecular docking studies. Information was collected from electronic databases, namely SciFinder, ScienceDirect, PubMed and Web of Science. Cannabis is mainly popular for its recreational use, but it is also traditionally used as remedy for the treatment of several diseases, including diabetes, digestive, circulatory, genital, nervous, urinary, skin and respiratory diseases. These biological proprieties are mainly due to the presence of bioactive metabolites represented by more than 550 different molecules. Molecular docking simulations proved the presence of affinities between Cannabis compounds and several enzymes responsible for anti-inflammatory, antidiabetic, antiepileptic and anticancer activities. Several biological activities have been evaluated on the metabolites of Cannabis sativa, and these works have shown the presence of antioxidant, antibacterial, anticoagulant, antifungal, anti-aflatoxigenic, insecticidal, anti-inflammatory, anticancer, neuroprotective and dermocosmetic activities. This paper presents the up-to-date reported investigations and opens many reflections and further research perspectives.

Key messageThree novel transcription factors were successfully identified and shown to interact with the trichome-specific THCAS promoter regulatory region.Cannabinoids are important secondary metabolites present in Cannabis sativa L. (cannabis). One cannabinoid that has received considerable attention, 9-tetrahydrocannabinol (THC), is derived from Delta-9-Tetrahydrocannabinolic acid (THCA) and responsible for the mood-altering and pain-relieving effects of cannabis. A detailed understanding of transcriptional control of THCA synthase (THCAS) is currently lacking. The primary site of cannabinoid biosynthesis is the glandular trichomes that form on female flowers. Transcription factors (TFs) have been shown to play an important role in secondary-metabolite biosynthesis and glandular trichome formation in Artemisia annua, Solanum lycopersicum and Humulus lupulus. However, analogous information is not available for cannabis. Here, we characterize a 548 bp fragment of the THCAS promoter and regulatory region that drives trichome-specific expression. Using this promoter fragment in a yeast-one-hybrid screen, we identified 3 novel TFs (CsAP2L1, CsWRKY1 and CsMYB1) and provided evidence that these 3 TFs regulate the THCAS promoter in planta. The O-Box element within the proximal region of the THCAS promoter is necessary for CsAP2L1-induced transcriptional activation of THCAS promoter. Similar to THCAS, the genes for all three TFs have trichome-specific expression, and subcellular localization of the TFs indicates that all three proteins are in the nucleus. CsAP2L1 and THCAS exhibit a similar temporal, spatial and strain-specific gene expression profiles, while those expression patterns of CsWRKY1 and CsMYB1 are opposite from THCAS. Our results identify CsAP2L1 playing a positive role in the regulation of THCAS expression, while CsWRKY1 and CsMYB1 may serve as negative regulators of THCAS expression.

Debates over Cannabis sativa L. and C. indica Lam. center on their taxonomic circumscription and rank. This perennial puzzle has been compounded by the viral spread of a vernacular nomenclature, \"Sativa\" and \"Indica,\" which does not correlate with C. sativa and C. indica. Ambiguities also envelop the epithets of wildtype Cannabis: the spontanea versus ruderalis debate (i.e., vernacular \"Ruderalis\"), as well as another pair of Cannabis epithets, afghanica and kafirstanica. To trace the rise of vernacular nomenclature, we begin with the protologues (original descriptions, synonymies, type specimens) of C. sativa and C. indica. Biogeographical evidence (obtained from the literature and herbarium specimens) suggests 18th—19th century botanists were biased in their assignment of these taxa to field specimens. This skewed the perception of Cannabis biodiversity and distribution. The development of vernacular \"Sativa,\" \"Indica,\" and \"Ruderalis\" was abetted by twentieth century botanists, who ignored original protologues and harbored their own cultural biases. Predominant taxonomic models by Vavilov, Small, Schultes, de Meijer, and Hillig are compared and critiqued. Small's model adheres closest to protologue data (with C.indica treated as a subspecies). \"Sativa\" and \"Indica\" are subpopulations of C. sativa subsp. indica; \"Ruderalis\" represents a protean assortment of plants, including C. sativa subsp. sativa and recent hybrids.

The cultivation of medical cannabis ( Cannabis sativa L.) is expanding in controlled environments, driven by evolving governmental regulations for healthcare supply. Increasing inflorescence weight and plant specialized metabolite (PSM) concentrations is critical, alongside maintaining product consistency. Medical cannabis is grown under different spectra and photosynthetic photon flux densities (PPFD), the interaction between spectrum and PPFD on inflorescence weight and PSM attracts attention by both industrialists and scientists. Plants were grown in climate-controlled rooms without solar light, where four spectra were applied: two low-white spectra (7B-20G-73R/Narrow and 6B-19G-75R/2Peaks), and two high-white (15B-42G-43R/Narrow and 17B-40G-43R/Broad) spectra. The low-white spectra differed in red wavelength peaks (100% 660 nm, versus 50:50% of 640:660 nm), the high-white spectra differed in spectrum broadness. All four spectra were applied at 600 and 1200 μmol m -2 s -1 . Irrespective of PPFD, white light with a dual red peak of 640 and 660 nm (6B-19G-75R/2Peaks) increased inflorescence weight, compared to white light with a single red peak of 660 nm (7B-20G-73R/Narrow) (tested at P = 0.1); this was associated with higher total plant dry matter production and a more open plant architecture, which likely enhanced light capture. At high PPFD, increasing white fraction and spectrum broadness (17B-40G-43R/Broad) produced similar inflorescence weights compared to white light with a dual red peak of 640 and 660 nm (6B-19G-75R/2Peaks). This was caused by an increase of both plant dry matter production and dry matter partitioning to the inflorescences. No spectrum or PPFD effects on cannabinoid concentrations were observed, although at high PPFD white light with a dual red peak of 640 and 660 nm (6B-19G-75R/2Peaks) increased terpenoid concentrations compared to the other spectra. At low PPFD, the combination of white light with 640 and 660 nm increased photosynthetic efficiency compared with white light with a single red peak of 660nm, indicating potential benefits in light use efficiency and promoting plant dry matter production. These results indicate that the interaction between spectrum and PPFD influences plant dry matter production. Dividing the light energy in the red waveband over both 640 and 660 nm equally shows potential in enhancing photosynthesis and plant dry matter production.

Light spectrum plays a crucial role in regulating the growth of hemp ( Cannabis sativa L.) plants and the biosynthesis of secondary metabolites. Several studies have demonstrated that additional red-light exposure increases biomass accumulation, while supplementary UV-A light stimulates cannabinoid synthesis. Nevertheless, the potential of stage-specific supplementation of red and UV-A light remains underexplored in its capacity to optimize cannabinoid yield in indoor hemp cultivation. In the present study, the effect of red light in combination with UV-A light on hemp biomass and cannabinoid accumulation was investigated using a high-CBD strain. There were four treatments: (1) white light throughout the growth period (control; V W R W ); (2) red light supplementation during the vegetative stage (V WR R W ); (3) UV-A supplementation (V W R WUV ) during the flowering stage; and (4) combined red and UV-A supplementation (V WR R WUV ) during the vegetative and flowering stages. Results showed that V WR R W promoted the number of effective branches (increased by 18.0%) compared to the control (V W R W ), resulting in an increase in inflorescence yield by 17.9%. V W R WUV increased CBG and CBD content by 52.7% and 12.1%, respectively, relative to the control. The effect of V WR R WUV on biomass and cannabinoid accumulation was the strongest among the treatments, with CBG and CBD yields reaching 0.53 g and 4.62 g per plant, representing significant increase of 91.8% ( p  < 0.01) and 44.1% ( p  < 0.01), respectively, compared to the control. However, there were no significant differences in CBD yield among the V WR R W , V W R WUV and V WR R WUV treatments, indicating that the combined supplementation of red and UV-A light did not have an additive effect on CBD accumulation. These findings highlight the potential of stage-specific spectral strategy to optimize both plant growth and phytochemical quantity.

In the field of Cannabis sativa L. derived products, the attention has been usually focused on plant inflorescences as a source of cannabinoids, with stems and seeds also utilized, while leaves are typically discharged. Although previous studies have examined the composition of C. sativa leaves to evaluate their antioxidant potential, their full value remains largely unknown. In this perspective, this study aims to reveal the hidden potential of C. sativa leaves to reposition them from a waste material to an alternative source of bioactive compounds. To this purpose, the leaves of four non-psychotropic C. sativa (hemp) varieties, including a cannabinoid-free one, were comprehensively characterized for their composition for the first time. The most relevant outcome of the present work was the first identification and characterization of squalene in hemp leaves, along with the development of a suitable extraction and analytical method for this bioactive triterpenoid. Among cannabinoids, cannabinoic acids were the most abundant compounds, complemented by the detection of several minor compounds having either the cannabidiol (CBD) or cannabigerol (CBG) scaffold. As for the other chemical constituents, cannflavins were the predominant non-cannabinoid phenolic compounds. The analysis also addressed to policosanols and terpenes, revealing some variety-specific volatile compounds, beside the most common ones. Overall, hemp leaves showed a rich chemical composition to be exploited in the pharmaceutical field, perfectly aligning with a circular economy perspective.

The optimization of nitrogen nutrition is critical for standardizing the production of bioactive compounds in medicinal Cannabis sativa L. Although deep-water culture (DWC) systems offer precise control over nutrient delivery, the specific effects of ammonium-to-nitrate (NH 4 + :NO 3 − ) ratios and nutrient solution strength on the phytochemical composition of cannabis flowers remain poorly understood. Notably, this study is the first to jointly examine the combined influence of the NH 4 + :NO 3 − ratio and solution strength in a DWC system for cannabis. This study investigated the synergistic impact of different NH 4 + :NO 3 − ratios and solution strength on morphological traits, phenolic content, antioxidant activity, and cannabinoid profile in organically grown Cannabis sativa L. flowers in a DWC system. Plants were subjected to varying NH 4 + :NO 3 − ratios (20:80, 40:60, 60:40) at full and half solution strengths. Morphological traits were measured at harvest. Methanolic extracts of flowers were analyzed for total phenolic (TPC) and flavonoid (TFC) content, antioxidant activity (DPPH assay), and major cannabinoids (GC-MS). Biomass production was maximized at a 40:60 ratio (full strength) and a 60:40 ratio (half strength). However, the highest TPC, TFC, and antioxidant activity were recorded at a 60:40 ratio under half solution strength, indicating a stress-induced upregulation of secondary metabolites. Cannabinoid synthesis was most favorable at a 40:60 ratio with full solution strength, yielding the highest levels of delta-9-THC (28.46%) and CBD (10.09%). A 60:40 ratio at full strength completely suppressed cannabinoid production, demonstrating acute ammonium toxicity. Cannabis phytochemistry is strongly influenced by both nitrogen form and nutrient concentration. A balanced NH 4 + :NO 3 − ratio (40:60) under full nutrition optimizes cannabinoid production, while a high NH 4 + ratio (60:40) under nutrient stress enhances phenolic antioxidants. These findings provide a precise framework for tailoring nutrient regimens to selectively target specific bioactive compound classes in medicinal cannabis production.

The use of the cannabis plant as a source of therapeutic compounds is gaining great importance since restrictions on its growth and use are gradually reduced throughout the world. Intensification of medical (drug type) cannabis production stimulated breeding activities aimed at developing new, improved cultivars with precisely defined, and stable cannabinoid profiles. The effects of several exogenous substances, known to be involved in sex expressions, such as silver thiosulfate (STS), gibberellic acid (GA), and colloidal silver, were analyzed in this study. Various concentrations were tested within 23 different treatments on two high cannabidiol (CBD) breeding populations. Our results showed that spraying whole plants with STS once is more efficient than the application of STS on shoot tips while spraying plants with 0.01% GA and intensive cutting is ineffective in stimulating the production of male flowers. Additionally, spraying whole plants with colloidal silver was also shown to be effective in the induction of male flowers on female plants, since it produced up to 379 male flowers per plant. The viability and fertility of the induced male flowers were confirmed by fluorescein diacetate (FDA) staining of pollen grains, in vitro and in vivo germination tests of pollen, counting the number of seeds developed after hybridization, and evaluating germination rates of developed seeds. Finally, one established protocol was implemented for crossing selected female plants. The cannabinoid profile of the progeny was compared with the profile of the parental population and an improvement in the biochemical profile of the breeding population was confirmed. The progeny had a higher and more uniform total CBD (tCBD) to total tetrahydrocannabinol (tTHC) ratio (up to 29.6; average 21.33 ± 0.39) compared with the original population (up to 18.8; average 7.83 ± 1.03). This is the first comprehensive report on the induction of fertile male flowers on female plants from dioecious medical cannabis ( Cannabis sativa L.).

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.