Integrated Transcriptomic and Metabolomic Analysis Reveal the Dynamic Process of Bama Hemp Seed Development and the Accumulation Mechanism of α-Linolenic Acid and Linoleic Acid.

Bama County is a world-famous longevity county in the Guangxi Province, China. Bama hemp is a traditional seed used in hemp cultivation in the Bama County. The seeds contain abundant unsaturated fatty acids, particularly linoleic acid (LA) and linolenic acid in the golden ratio. These two substances have been proven to be related to human health and the prevention of various diseases. However, the seed development and seed oil accumulation mechanisms remain unclear. This study employed a combined analysis of physiological, transcriptomic, and metabolomic parameters to elucidate the fatty acid formation patterns in Bama hemp seeds throughout development. We found that seed oil accumulated at a late stage in embryo development, with seed oil accumulation following an "S″-shaped growth curve, and positively correlated with seed size, sugar content, protein content, and starch content. Transcriptome analysis identified genes related to the metabolism of LA, α-linolenic acid (ALA), and jasmonic acid (JA). We found that the FAD2 gene was upregulated 165.26 folds and the FAD3 gene was downregulated 6.15 folds at day 21. Metabolomic changes in LA, ALA, and JA compounds suggested a competitive relationship among these substances. Our findings indicate that the peak period of substance accumulation and nutrient accumulation in Bama hemp seeds occurs during the midstage of seed development (day 21) rather than in the late stage (day 40). The results of this research will provide a theoretical basis for local cultivation and deep processing of Bama hemp.

Deep learning-based quantification and transcriptomic profiling reveal a methyl jasmonate-mediated glandular trichome formation pathway in Cannabis sativa.

Cannabis glandular trichomes (GTs) are economically and biotechnologically important structures that have a remarkable morphology and capacity to produce, store, and secrete diverse classes of secondary metabolites. However, our understanding of the developmental changes and the underlying molecular processes involved in cannabis GT development is limited. In this study, we developed Cannabis Glandular Trichome Detection Model (CGTDM), a deep learning-based model capable of differentiating and quantifying three types of cannabis GTs with a high degree of efficiency and accuracy. By profiling at eight different time points, we captured dynamic changes in gene expression, phenotypes, and metabolic processes associated with GT development. By integrating weighted gene co-expression network analysis with CGTDM measurements, we established correlations between phenotypic variations in GT traits and the global transcriptome profiles across the developmental gradient. Notably, we identified a module containing methyl jasmonate (MeJA)-responsive genes that significantly correlated with stalked GT density and cannabinoid content during development, suggesting the existence of a MeJA-mediated GT formation pathway. Our findings were further supported by the successful promotion of GT development in cannabis through exogenous MeJA treatment. Importantly, we have identified CsMYC4 as a key transcription factor that positively regulates GT formation via MeJA signaling in cannabis. These findings provide novel tools for GT detection and counting, as well as valuable information for understanding the molecular regulatory mechanism of GT formation, which has the potential to facilitate the molecular breeding, targeted engineering, informed harvest timing, and manipulation of cannabinoid production.

Impacto de la legalización del consumo recreativo del cannabis / Impact of the legalisation of recreational cannabis use

En los últimos años se han producido importantes cambios legislativos en numerosos países respecto al consumo de cannabis con fines medicinales y/o recreativos, que han facilitado su accesibilidad. Actualmente, Uruguay, Canadá y algunos estados de EE.UU. han legalizado el consumo recreativo, aplicando distintos modelos legislativos. El objetivo de la presente revisión es analizar los efectos que ha tenido la legalización del cannabis recreativo sobre su consumo y sus consecuencias. En general, las evidencias indican que la legalización se ha asociado a un descenso en el precio, mayor concentración de THC (potencia), mayor diversidad de presentaciones para su consumo, una menor percepción de riesgo y un incremento en el consumo en adultos y de forma moderada en adolescentes (aunque sea ilegal el consumo para ellos), así como un aumento de las consecuencias adversas derivadas del consumo en la salud pública. Se ha producido un descenso en los arrestos relacionados con el consumo, pero el mercado ilegal sigue utilizándose de forma habitual. No se ha detectado un incremento de la demanda de tratamiento por este consumo. Por el momento, estos cambios legislativos no han conseguido alcanzar sus objetivos principales que eran suprimir el mercado ilegal y proteger a los grupos más vulnerables, mientras que, por el contrario, parecen implicar un incremento de algunos aspectos negativos asociados al consumo de cannabis. Sin embargo, teniendo en cuenta que la mayoría de estos cambios legislativos han entrado en vigor hace relativamente poco tiempo, se requiere un periodo de seguimiento mayor para poder extraer conclusiones definitivas. (AU)

In recent years, there have been important legislative changes in many countries regarding the use of cannabis for medicinal and/or recreational purposes, which have facilitated access to it. Uruguay, Canada and some of the US states are the only jurisdictions that have legalised recreational consumption, applying different legislative models. The aim of this review is to analyse the effects that the legalisation of recreational cannabis has had on its use and its consequences. In general, the evidence accumulated to date indicates that the legalisation of cannabis has been associated with a decrease in the price of the substance, higher concentration of THC (potency), greater diversity of presentations for consumption, lower risk perception and an increase in consumption in adults and moderately in adolescents (even though it is illegal for them to consume), as well as an increase in the adverse consequences derived from cannabis consumption on public health. There has been a decrease in drug-related arrests, but the illegal market continues to be frequently used. No increase in the demand for treatment due to cannabis consumption has been detected. Therefore, these legislative changes have so far failed to achieve their main objectives, which were to suppress the illegal market and protect the most vulnerable groups, while on the contrary, they seem to imply an increase in some of the negative aspects associated with cannabis consumption. However, taking into account that most of these legislative changes have entered into force relatively recently, a longer follow-up period is required to be able to draw definitive conclusions. (AU)

Genome-wide identification and expression analysis response to GA3 stresses of WRKY gene family in seed hemp (Cannabis sativa L).

WRKY transcription factor is one of the largest transcription factor families in higher plants. However, the investigations of the WRKY gene family have not yet been reported in seed hemp. In the present study, we identified 39 CasWRKYs at the genome-wide level and analyzed phylogenetic relationship, chromosome location, cis-acting elements, gene structure, conserved motif, and expression pattern. Based on the gene structure and phylogenetic analyses, CasWRKY proteins were divided into 3 groups and 7 subgroups. The gene duplication investigation revealed that 6 and 5 pairs of CasWRKY genes underwent tandem and segmental duplication events, respectively. These events may contribute to the diversity and expansion of the CasWRKY gene family. The regulatory elements in the promoter regions of CasWRKYs contained diverse cis-regulatory elements, among which P-box cis-regulatory elements showed high frequency, indicating that CasWRKYs can respond to the regulation of gibberellin. The expression profiles derived from RNA-seq and qRT-PCR showed that 13 CasWRKY genes could respond to GA3 stress and affect fiber development, as well as play significant roles in stem growth and development. This study will serve as molecular basis and practical reference for further exploring the genetic evolution and biological function of CasWRKY genes in seed hemp.

Endocannabinoid system contributions to sex-specific adolescent neurodevelopment.

With an increasing number of countries and states adopting legislation permitting the use of cannabis for medical purposes, there is a growing interest among health and research professionals into the system through which cannabinoids principally act, the endocannabinoid system (ECS). Much of the seminal research into the ECS dates back only 30 years and, although there has been tremendous development within the field during this time, many questions remain. More recently, investigations have emerged examining the contributions of the ECS to normative development and the effect of altering this system during important critical periods. One such period is adolescence, a unique period during which brain and behaviours are maturing and reorganizing in preparation for adulthood, including shifts in endocannabinoid biology. The purpose of this review is to discuss findings to date regarding the maturation of the ECS during adolescence and the consequences of manipulations of the ECS during this period to normative neurodevelopmental processes, as well as highlight sex differences in ECS function, important technical considerations, and future directions. Because most of what we know is derived from preclinical studies on rodents, we provide relevant background of this model and some commentary on the translational relevance of the research in this area.

Assessment of the Cannabinoid Content from Different Varieties of Cannabis sativa L. during the Growth Stages in Three Regions.

Hemp (Cannabis sativa L.) belongs to the Cannabaceae family. It is very rich in chemical constituents, especially the cannabinoids which has not been reported in any other plant, and has broad pharmacological properties. Hemp as a multi-purpose crop is a good source of fibers, seed, fixed and volatile oil. It is known that the cannabinoid content of hemp is related to genetic factors, as well as plant's growth stages and environmental factors such as latitude, altitude, weather, particularly moisture availability and nutrient supply during the growing season. The present study was designed to produce hemp that contains allowable concentration of THC (<3 %) by comparing different varieties of hemp, different stages of plant growth, and different geographical locations where it was planted. To achieve this, seeds of two native populations from Iran (Fars and Yazd Provinces) and one foreign variety from France (Fedora17, as an industrial hemp cultivar) with its progenies (Fedora17-2) were cultivated in 3 research fields (Gilan, Golestan and Alborz provinces) in Iran. The following plant materials were extracted with methanol/chloroform and analyzed by HPLC: foliage in the vegetative stage, inflorescent in the flowering stage, inflorescent of seeds in the seeding stage and the mature seed. The THC concentration of Fedora17 (Fed17) in all three geographical locations was found to be under 0.03 % or even non-detectable. Same result was also observed in its progenies (Fed17-2), indicating stability of the trait in this cultivar. The THC concentration of the Yazd variety that was planted in Alborz and Gilan regions was less than 0.080 % in all growth stages. The female flowers planted in Golestan, showed a THC concentration of 1.029 % which was more than the allowed THC concentration of <3 %. The THC concentration in all growth stages of all of the different varieties planted varied from 0 to 1.392 %. The above results indicates that the type of cannabinoid produced depends on the difference in genetic prosperities of the different seed types as well as the growth stage in which the plant material was extracted. On the other hand, the climate and the region in which the seeds were planted had little influence on the THC concentration.

Distribution of trespass cannabis cultivation and its risk to sensitive forest predators in California and Southern Oregon.

Illegal cannabis cultivation on public lands has emerged as a major threat to wildlife in California and southern Oregon due to the rampant use of pesticides, habitat destruction, and water diversions associated with trespass grow sites. The spatial distribution of cultivation sites, and the factors influencing where they are placed, remain largely unknown due to covert siting practices and limited surveillance funding. We obtained cannabis grow-site locality data from law enforcement agencies and used them to model the potential distribution of cultivation sites in forested regions of California and southern Oregon using maximum entropy (MaxEnt) methods. We mapped the likely distribution of trespass cannabis cultivation sites and identified environmental variables influencing where growers establish their plots to better understand the cumulative impacts of trespass cannabis cultivation on wildlife. We overlaid the resulting grow-site risk maps with habitat distribution maps for three forest species of conservation concern: Pacific fisher (Pekania pennanti), Humboldt marten (Martes caurina humboldtensis), and northern spotted owl (Strix occidentalis caurina). Results indicate that cannabis cultivation is fairly predictably distributed on public lands in low to mid-elevation (~800-1600m) forests and on moderate slopes (~30-60%). Somewhat paradoxically, results also suggest that growers either preferred sites inside of recently disturbed vegetation (especially those burned 8-12 years prior to cultivation) or well outside (>500m) of recent disturbance, perhaps indicating avoidance of open edges. We ground-truthed the model by surveying randomly selected stream courses for cultivation site presence in subsets of the modeling region and found previously undiscovered sites mostly within areas with predicted high likelihood of grow-site occurrence. Moderate to high-likelihood areas of trespass cultivation overlapped with 40 to 48% of modeled habitats of the three sensitive species. For the endangered southern Sierra Nevada fisher population, moderate-high likelihood growing areas overlapped with over 37% of modeled fisher denning habitat and with 100% of annual female fisher home ranges (mean overlap = 48.0% + 27.0 SD; n = 134) in two intensively studied populations on the Sierra National Forest. Locating and reclaiming contaminated cannabis grow sites by removing all environmental contaminants should be a high priority for resource managers.

Recalcitrance of Cannabis sativa to de novo regeneration; a multi-genotype replication study.

Cannabis sativa is relatively recalcitrant to de novo regeneration, but several studies have reported shoot organogenesis or somatic embryogenesis from non-meristematic tissues. Most report infrequent regeneration rates from these tissues, but a landmark publication from 2010 achieved regeneration from leaf explants with a 96% response rate, producing an average of 12.3 shoots per explant in a single drug-type accession. Despite the importance regeneration plays in plant biotechnology and the renewed interest in this crop the aforementioned protocol has not been used in subsequent papers in the decade since it was published, raising concerns over its reproducibility. Here we attempted to replicate this important Cannabis regeneration study and expand the original scope of the study by testing it across 10 drug-type C. sativa genotypes to assess genotypic variation. In our study, callus was induced in all 10 genotypes but callus growth and appearance substantially differed among cultivars, with the most responsive genotype producing 6-fold more callus than the least responsive. The shoot induction medium failed to induce shoot organogenesis in any of the 10 cultivars tested, instead resulting in necrosis of the calli. The findings of this replication study raise concerns about the replicability of existing methods. However, some details of the protocol could not be replicated due to missing details in the original paper and regulatory issues, which could have impacted the outcome. These results highlight the importance of using multiple genotypes in such studies and providing detailed methods to facilitate replication.

Yield, Characterization, and Possible Exploitation of Cannabis Sativa L. Roots Grown under Aeroponics Cultivation.

Cannabis sativa L. has been used for a long time to obtain food, fiber, and as a medicinal and psychoactive plant. Today, the nutraceutical potential of C.sativa is being increasingly reappraised; however, C. sativa roots remain poorly studied, despite citations in the scientific literature. In this direction, we identified and quantified the presence of valuable bioactives (namely, ß-sitosterol, stigmasterol, campesterol, friedelin, and epi-friedelanol) in the root extracts of C. sativa, a finding which might pave the way to the exploitation of the therapeutic potential of all parts of the C. sativa plant. To facilitate root harvesting and processing, aeroponic (AP) and aeroponic-elicited cultures (AEP) were established and compared to soil-cultivated plants (SP). Interestingly, considerably increased plant growth-particularly of the roots-and a significant increase (up to 20-fold in the case of ß-sitosterol) in the total content of the aforementioned roots' bioactive molecules were observed in AP and AEP. In conclusion, aeroponics, an easy, standardized, contaminant-free cultivation technique, facilitates the harvesting/processing of roots along with a greater production of their secondary bioactive metabolites, which could be utilized in the formulation of health-promoting and health-care products.

Response of essential oil hemp (Cannabis sativa L.) growth, biomass, and cannabinoid profiles to varying fertigation rates.

Five essential oil hemp (Cannabis sativa L.) cultivars (Cherry Blossom, Cherry Blossom (Tuan), Berry Blossom, Cherry Wine, and Cherry Blossom × Trump) were treated with six fertigation treatments to quantify the effects of synthetic fertilizer rates and irrigation electrical conductivity on plant growth, biomass accumulation, and cannabinoid profiles. Irrigation water was injected with a commercial 20-20-20 fertilizer at rates of 0, 50, 150, 300, 450, and 600 ppm nitrogen equating to 0.33 (control), 0.54, 0.96, 1.59, 2.22, and 2.85 dS m-1, respectively. Plants were grown under artificial lighting (18 hr) to maintain vegetative growth for eight weeks, followed by an eight-week flowering period. High linear relationship between chlorophyll concentrations and SPAD-502 measurements validated the utilization of SPAD meters to rapidly identify nutrient deficiency in essential oil hemp. Cultivars expressed significant variation in plant height and cannabinoid profiles (% dry mass), in concurrence with limited biomass and cannabinoid (g per plant) yield variation. Cherry Blossom was the best performing cultivar and Cherry Wine was the least productive. Variation in plant growth, biomass, and cannabinoid concentrations were affected to a greater extent by fertilizer rates. Optimal fertilizer rates were observed at 50 ppm N, while increased fertilizer rates significantly reduced plant growth, biomass accumulation, and cannabinoid concentrations. Increased fertilizer rates (> 300 ppm N) resulted in compliant THC levels (< 0.3%), although when coupled with biomass reductions resulted in minimal cannabinoid yields. Additionally, CBD concentration demonstrated higher sensitivity to increased fertilizer rates (> 300 ppm N) compared to THC and CBG (> 450 ppm N). The results of this study can serve as a guide when using fertigation methods on essential oil hemp cultivars; although results may differ with cultivar selection, environmental conditions, and management practices.

Photons from NIR LEDs can delay flowering in short-day soybean and Cannabis: Implications for phytochrome activity.

Photons during the dark period delay flowering in short-day plants (SDP). Red photons applied at night convert phytochromes to the active far-red absorbing form (Pfr), leading to inhibition of flowering. Far-red photons (greater than 700 nm) re-induce flowering when applied after a pulse of red photons during the dark period. However, far-red photons at sufficiently high intensity and duration delay flowering in sensitive species. Mechanistically, this response occurs because phytochrome-red (Pr) absorbance is not zero beyond 700 nm. We applied nighttime photons from near infrared (NIR) LEDs (peak 850 nm) over a 12 h dark period. Flowering was delayed in Glycine max and Cannabis sativa (two photosensitive species) by 3 and 12 days, respectively, as the flux of photons from NIR LEDs was increased up to 83 and 116 µmol m-2 s-1. This suggests that long wavelength photons from NIR LEDs can activate phytochromes (convert Pr to Pfr) and thus alter plant development.

Essential Oil of Cannabis sativa L: Comparison of Yield and Chemical Composition of 11 Hemp Genotypes.

Cannabis sativa L. is an annual species cultivated since antiquity for different purposes. While, in the past, hemp inflorescences were considered crop residues, at present, they are regarded as valuable raw materials with different applications, among which extraction of the essential oil (EO) has gained increasing interest in many fields. The aim of the present study is the evaluation of the yield and the chemical composition of the EO obtained by hydrodistillation from eleven hemp genotypes, cultivated in the same location for two consecutive growing seasons. The composition of the EOs was analyzed by GC-MS, and then subjected to multivariate statistical analysis. Sesquiterpenes represented the main class of compounds in all the EOs, both in their hydrocarbon and oxygenated forms, with relative abundances ranging from 47.1 to 78.5%; the only exception was the Felina 32 sample collected in 2019, in which cannabinoids predominated. Cannabinoids were the second most abundant class of compounds, of which cannabidiol was the main one, with relative abundances between 11.8 and 51.5%. The statistical distribution of the samples, performed on the complete chemical composition of the EOs, evidenced a partition based on the year of cultivation, rather than on the genotype, with the exception of Uso-31. Regarding the extraction yield, a significant variation was evidenced among both the genotypes and the years of cultivation.

Quantitative trait loci controlling agronomic and biochemical traits in Cannabis sativa.

Understanding the genetic basis of complex traits is a fundamental goal of evolutionary genetics. Yet, the genetics controlling complex traits in many important species such as hemp (Cannabis sativa) remain poorly investigated. Because hemp's change in legal status with the 2014 and 2018 U.S. Federal Farm Bills, interest in the genetics controlling its numerous agriculturally important traits has steadily increased. To better understand the genetics of agriculturally important traits in hemp, we developed an F2 population by crossing two phenotypically distinct hemp cultivars (Carmagnola and USO31). Using whole-genome sequencing, we mapped quantitative trait loci (QTL) associated with variation in numerous agronomic and biochemical traits. A total of 69 loci associated with agronomic (34) and biochemical (35) trait variation were identified. We found that most QTL co-localized, suggesting that the phenotypic distinctions between Carmagnola and USO31 are largely controlled by a small number of loci. We identified TINY and olivetol synthase as candidate genes underlying co-localized QTL clusters for agronomic and biochemical traits, respectively. We functionally validated the olivetol synthase candidate by expressing the alleles in yeast. Gas chromatography-mass spectrometry assays of extracts from these yeast colonies suggest that the USO31 olivetol synthase is functionally less active and potentially explains why USO31 produces lower cannabinoids compared to Carmagnola. Overall, our results help modernize the genomic understanding of complex traits in hemp.

A Multimethodological Characterization of Cannabis sativa L. Inflorescences from Seven Dioecious Cultivars Grown in Italy: The Effect of Different Harvesting Stages.

The chemical profile of the female inflorescence extracts from seven Cannabis sativa L. dioecious cultivars (Carmagnola, Fibranova, Eletta Campana, Antal, Tiborszallasi, Kompolti, and Tisza) was monitored at three harvesting stages (4, 14, and 30 September), reaching from the beginning of flowering to end of flowering/beginning of seed formation, using untargeted nuclear magnetic resonance (NMR) and targeted (ultra-high-performance liquid chromatography (UHPLC) and spectrophotometry) analyses. The tetrahydrocannabinol content was always below the legal limits (<0.6%) in all the analyzed samples. The NMR metabolite profile (sugars, organic acids, amino acids, and minor compounds) subjected to principal components analysis (PCA) showed a strong variability according to the harvesting stages: samples harvested in stage I were characterized by a high content of sucrose and myo-inositol, whereas the ones harvested in stage II showed high levels of succinic acid, alanine, valine, isoleucine, phenylalanine, and threonine. Samples harvested in stage III were characterized by high levels of glucose, fructose, choline, trigonelline, malic acid, formic acid, and some amino acids. The ratio between chlorophylls and carotenoids content indicated that all plants grew up exposed to the sun, the Eletta Campana cultivar having the highest pigment amount. Tiborszallasi cultivar showed the highest polyphenol content. The highest antioxidant activity was generally observed in stage II. All these results suggested that the Cannabis sativa L. inflorescences of each analyzed dioecious hemp cultivar presented a peculiar chemical profile affected by the harvesting stage. This information could be useful for producers and industries to harvest inflorescences in the appropriate stage to obtain samples with a peculiar chemical profile suitable for proper applications.

The characterization of key physiological traits of medicinal cannabis (Cannabis sativa L.) as a tool for precision breeding.

BACKGROUND: For millennia, drug-type cannabis strains were extensively used for various medicinal, ritual, and inebriant applications. However, cannabis prohibition during the last century led to cultivation and breeding activities being conducted under clandestine conditions, while scientific development of the crop ceased. Recently, the potential of medicinal cannabis has been reacknowledged and the now expanding industry requires optimal and scientifically characterized varieties. However, scientific knowledge that can propel this advancement is sorely lacking. To address this issue, the current study aims to provide a better understanding of key physiological and phenological traits that can facilitate the breeding of advanced cultivars. RESULTS: A diverse population of 121 genotypes of high-THC or balanced THC-CBD ratio was cultivated under a controlled environment facility and 13 plant parameters were measured. No physiological association across genotypes attributed to the same vernacular classification was observed. Floral bud dry weight was found to be positively associated with plant height and stem diameter but not with days to maturation. Furthermore, the heritability of both plant height and days to maturation was relatively high, but for plant height it decreased during the vegetative growth phase. To advance breeding efficacy, a prediction equation for forecasting floral bud dry weight was generated, driven by parameters that can be detected during the vegetative growth phase solely. CONCLUSIONS: Our findings suggest that selection for taller and fast-growing genotypes is likely to lead to an increase in floral bud productivity. It was also found that the final plant height and stem diameter are determined by 5 independent factors that can be used to maximize productivity through cultivation adjustments. The proposed prediction equation can facilitate the selection of prolific genotypes without the completion of a full cultivation cycle. Future studies that will associate genome-wide variation with plants morphological traits and cannabinoid profile will enable precise and accelerated breeding through genomic selection approaches.

Neuropsychological and neurophysiological predictors and consequences of cannabis and illicit substance use during neurodevelopment: a systematic review of longitudinal studies.

Adolescence and early adulthood are crucial periods of neurodevelopment characterised by functional, structural, and cognitive maturation, which helps prepare young people for adulthood. This systematic review of longitudinal studies aims to delineate neural predictors from neural consequences of cannabis and illicit substance use, as well as investigate the potential for the developing brain (at ages 10-25 years) to recover after damage. Five databases were searched to yield a total of 38 eligible studies, with some assessing multiple outcome techniques, including 22 neuroimaging, two neurophysiological, and 22 neuropsychological findings. High-quality evidence suggested that delayed or irregular neurodevelopment in executive functioning, particularly emotional perception, might predispose young people to higher frequency substance use. There was evidence of functional, structural, and cognitive deficits proceeding substance use, with harm potentially dependent on the frequency of use and recovery potentially dependent on the duration of use. Identifying aberrant neurodevelopment in young people is crucial for preventing substance use-related harm.

Biochemical aspects of seeds from Cannabis sativa L. plants grown in a mountain environment.

Cannabis sativa L. (hemp) is a versatile plant which can adapt to various environmental conditions. Hempseeds provide high quality lipids, mainly represented by polyunsaturated acids, and highly digestible proteins rich of essential aminoacids. Hempseed composition can vary according to plant genotype, but other factors such as agronomic and climatic conditions can affect the presence of nutraceutic compounds. In this research, seeds from two cultivars of C. sativa (Futura 75 and Finola) grown in a mountain environment of the Italian Alps were analyzed. The main purpose of this study was to investigate changes in the protein profile of seeds obtained from such environments, using two methods (sequential and total proteins) for protein extraction and two analytical approaches SDS-PAGE and 2D-gel electrophoresis, followed by protein identification by mass spectrometry. The fatty acids profile and carotenoids content were also analysed. Mountain environments mainly affected fatty acid and protein profiles of Finola seeds. These changes were not predictable by the sole comparison of certified seeds from Futura 75 and Finola cultivars. The fatty acid profile confirmed a high PUFA content in both cultivars from mountain area, while protein analysis revealed a decrease in the protein content of Finola seeds from the experimental fields.

Comparative RNA-Seq analysis reveals genes associated with masculinization in female Cannabis sativa.

MAIN CONCLUSION: Using RNA profiling, we identified several silver thiosulfate-induced genes that potentially control the masculinization of female Cannabis sativa plants. Genetically female Cannabis sativa plants normally bear female flowers, but can develop male flowers in response to environmental and developmental cues. In an attempt to elucidate the molecular elements responsible for sex expression in C. sativa plants, we developed genetically female lines producing both female and chemically-induced male flowers. Furthermore, we carried out RNA-Seq assays aimed at identifying differentially expressed genes responsible for male flower development in female plants. The results revealed over 10,500 differentially expressed genes, of which around 200 potentially control masculinization of female cannabis plants. These genes include transcription factors and other genes involved in male organ (i.e., anther and pollen) development, as well as genes involved in phytohormone signalling and male-biased phenotypes. The expressions of 15 of these genes were further validated by qPCR assay confirming similar expression patterns to that of RNA-Seq data. These genes would be useful for understanding predisposed plants producing flowers of both sex types in the same plant, and help breeders to regulate the masculinization of female plants through targeted breeding and plant biotechnology.

The Cannabis Spread throughout the Continents and its Therapeutic Use in History.

BACKGROUND: Cannabis sativa L. (C. sativa) is a plant whose use as a therapeutic agent shares its origins with the first Far East's human societies. Cannabis has been used not only for recreational purposes but as food to obtain textile fibers, to produce hemp paper, to treat many physical and mental disorders. AIM: This review aims to provide a complete assessment of the deep knowledge of the cannabis psychoactive effects and medicinal properties in the course of history covering i.) The empirical use of the seeds and the inflorescences to treat many physical ailments by the ancient Oriental physicians' ii.) The current use of cannabis as a therapeutic agent after the discovery of its key psychoactive constituent and the human endogenous endocannabinoid system. METHODS: This study was performed through a detailed analysis of the studies on the historical significance and medical applications of Cannabis sativa by using international scientific databases, historical and medical books, ancient Greek and Chinese manuscripts translations, library and statistical data from government reports and texts from the National Library of Greece (Stavros Niarchos Foundation), from the School of Health Sciences of the National and Kapodistrian University of Athens (Greece). We selected papers and texts focusing on a historical point of view about the medical importance of the plant and its applications for a therapeutic purpose in the past. RESULTS AND CONCLUSION: Through a detailed analysis of the available resources about the origins of C. sativa, we found that its use by ancient civilizations as a source of food and textile fibers dates back over 10,000 years, while its therapeutic applications have been improved over the centuries, from the ancient East medicine of the 2nd and 1st millennium B.C. to the more recent introduction in the Western world after the 1st century A.D. In the 20th and 21st centuries, Cannabis and its derivatives have been considered as a menace and banned throughout the world, but nowadays, they are still the most widely consumed illicit drugs all over the world. Its legalization in some jurisdictions has been accompanied by new lines of research to investigate its possible applications for medical and therapeutic purposes.