Drug-type Cannabis Research Articles

Sink strength, nutrient allocation, cannabinoid yield and associated transcript profiles vary in two drug-type Cannabis chemovars.

Cannabis sativa L. is one of the oldest domesticated crops. Hemp-type cultivars, which predominantly produce non-intoxicating cannabidiol (CBD), have been selected for their fast growth, seed, and fibre production, while drug-type chemovars were bred for high accumulation of tetrahydrocannabinol (THC). We investigated how the generation of CBD-dominant chemovars by introgression of hemp- into drug-type Cannabis impacted plant performance. The THC-dominant chemovar showed superior sink strength, higher flower biomass and demand-driven control of nutrient uptake. By contrast, the CBD-dominant chemovar hyperaccumulated phosphate in sink organs leading to reduced carbon and nitrogen assimilation in leaves, which limited flower biomass and cannabinoid yield. RNA-seq analyses determined organ- and chemovar-specific differences in expression of genes associated with nitrate and phosphate homeostasis as well as growth-regulating transcription factors that were correlated with measured traits. Among these were genes positively selected for during Cannabis domestication encoding an inhibitor of the phosphate starvation response SPX DOMAIN GENE3, nitrate reductase and two nitrate transporters. Altered nutrient sensing, acquisition or distribution are likely a consequence of adaption to growth on marginal, low-nutrient input lands in hemp. Our data provide evidence that such ancestral traits may become detrimental for female flower development and consequently overall CBD yield in protected cropping environments.

Same, Yet Different: Towards Understanding Nutrient Use In Hemp- And Drug-Type Cannabis.

Cannabis sativa L., one of humanity's oldest cultivated crops, has a complex domestication history due to its diverse uses for fibre, seed, oil and drugs, and its wide geographic distribution. This review explores how human selection has shaped the biology of hemp and drug-type Cannabis, focusing on acquisition and utilisation of nitrogen and phosphorus, and how resulting changes in source-sink relations shape their contrasting phenology. Hemp has been optimized for rapid, slender growth and nutrient efficiency, whereas drug-type cultivars have been selected for compact growth with large phytocannabinoid producing female inflorescences. Understanding these nutrient use and ontogenetic differences will enhance our general understanding of resource allocation in plants. Knowledge gained in comparison with other model species, such as tomato, rice or Arabidopsis thaliana can help inform crop improvement and sustainability in the Cannabis industry.

Phytocannabinoid profile and potency of cannabis resin (hashish) of northwest Himalayas of India.

Cannabis is one of the most consumed illicit drugs and the potency of cannabis products is of note due to health-related concerns. Hand-rubbed hashish is the ancient technique of extracting psychoactive resin from cannabis plants and is practiced in the Indian Himalayas. This study establishes the cannabinoid profile and potency of hand-rubbed hashish collected from 20 regions of the northwest Himalayas. Fifty-eight hashish samples were analyzed using a validated high-performance liquid chromatography-diode array detection (HPLC-DAD) method. Ten cannabinoids were quantified including acidic (THCA & CBDA), and neutral compounds (CBDV, THCV, CBD, CBG, CBN, Δ9-THC, Δ8-THC, and CBC). The mean concentration (w/w%) of Δ9-THC is 26%; THCA is 15% and THCTotal is 40% is observed in the studied hashish samples. The majority (70%) of the hashish samples were categorized in chemotype I with the THC:CBD:CBN ratio of 91:3:4, and the remaining 30% were categorized under chemotype II with the ratio of 76:15:8. Diverse qualities of hashish are produced in the studied regions as per the seed, plant selection, and skills of manual rubbing, which results in potency variations. The average difference between the least and highest potent hand-rubbed hashish of a region is 27 w/w% (THCTotal). The other studied non-psychoactive cannabinoids have a mean w/w% of <5%, followed by 6% of CBDA. It is concluded that the cultivated and wild cannabis fields in the northwest Himalayas belong to the drug-type cannabis subspecies. Hand-rubbed hashish holds traditional significance and impacts the current policies of legislation.

A versatile, rapid Agrobacterium-mediated transient expression system for functional genomics studies in cannabis seedling.

We have developed and optimized a rapid, versatile Agrobacterium-mediated transient expression system for cannabis seedlings that can be used in functional genomics studies of both hemp-type and drug-type cannabis. Cannabis (Cannabis sativa L.) holds great promise in the medical and food industries due to its diverse chemical composition, including specialized cannabinoids. However, the study of key genes involved in various biological processes, including secondary metabolite biosynthesis, has been hampered by the lack of efficient in vivo functional analysis methods. Here, we present a novel, short-cycle, high-efficiency transformation method for cannabis seedlings using Agrobacterium tumefaciens. We used the RUBY reporter system to monitor transformation results without the need for chemical treatments or specialized equipment. Four strains of A. tumefaciens (GV3101, EHA105, LBA4404, and AGL1) were evaluated for transformation efficiency, with LBA4404 and AGL1 showing superior performance. The versatility of the system was further demonstrated by successful transformation with GFP and GUS reporter genes. In addition, syringe infiltration was explored as an alternative to vacuum infiltration, offering simplicity and efficiency for high-throughput applications. Our method allows rapid and efficient in vivo transformation of cannabis seedlings, facilitating large-scale protein expression and high-throughput characterization studies.

Trends of the cannabis market in Poland over the last decade = Tendencje na rynku konopi indyjskich w ostatniej dekadzie w Polsce

Cannabis is the most widely used drug. in recent years, interest in cannabis has grown significantly. over the last decade, the number of cannabis users worldwide has increased by almost 21%, reaching approximately 219 million people in 2021. Can- nabis cultivation covers practically all regions of the world. In recent years, 151 countries have reported cannabis cultivation, both outdoors and indoors. The increased growth of the latter is particularly observed in North America and Europe. Global data shows that in 2021, 5,226 tons of marihuana and 2,014 tons of hashish were confiscated. an increase in seizures of can- nabis products is observed in Europe and Poland. Another trend concerns the increasing content of the psychoactive substance Δ9-tetrahydrocannabinol (Δ9-THC) in cannabis plants. According to UNODC data, over the last 25 years there has been a 4-fold increase in the content of Δ9-THc in cannabis. over the last decade, the average content of Δ9-tetrahydrocannabinol in marihua- na seized in europe has increased approximately twice, and in hashish three times. The aim of this work is to assess potential changes in the content of the main psychoactive ingredients in cannabis in Poland in the last decade (2012–2022), based on expert opinions developed by the Institute of Forensic research in Kraków. The conduct- ed analyses confirmed the trends observed around the world. in the last ten years, the number of cases in which the drug-type cannabis was detected increased from 5.3% to 68.4% of total number of illicit drug cases examined for the presence of narcotic drugs, psychotropic substances and new psychoactive substances. an increasing trend in the content of Δ9-THC in the drug-type cannabis was also noted. over the last decade, the average total content of Δ9-THC increased almost three times and in 2022 reached 12%. This is due to the growth of cannabis cultivation in Europe, carried out using the technique known as sinsemilla, genetic modifications, as well as the appearance on the market of the feminized cannabis seeds.

Genetic insights into agronomic and morphological traits of drug-type cannabis revealed by genome-wide association studies

Cannabis sativa L., previously concealed by prohibition, is now a versatile and promising plant, thanks to recent legalization, opening doors for medical research and industry growth. However, years of prohibition have left the Cannabis research community lagging behind in understanding Cannabis genetics and trait inheritance compared to other major crops. To address this gap, we conducted a comprehensive genome-wide association study (GWAS) of nine key agronomic and morphological traits, using a panel of 176 drug-type Cannabis accessions from the Canadian legal market. Utilizing high-density genotyping-by-sequencing (HD-GBS), we successfully generated dense genotyping data in Cannabis, resulting in a catalog of 800 K genetic variants, of which 282 K common variants were retained for GWAS analysis. Through GWAS analysis, we identified 18 markers significantly associated with agronomic and morphological traits. Several identified markers exert a substantial phenotypic impact, guided us to putative candidate genes that reside in high linkage-disequilibrium (LD) with the markers. These findings lay a solid foundation for an innovative cannabis research, leveraging genetic markers to inform breeding programs aimed at meeting diverse needs in the industry.

Loss of daylength sensitivity by splice site mutation in Cannabis pseudo-response regulator.

Photoperiod insensitivity (auto-flowering) in drug-type Cannabis sativa circumvents the need for short day (SD) flowering requirements making outdoor cultivation in high latitudes possible. However, the benefits of photoperiod insensitivity are counterbalanced by low cannabinoid content and poor flower quality in auto-flowering genotypes. Despite recent studies in cannabis flowering, a mechanistic understanding of photoperiod insensitivity is still lacking. We used a combination of genome-wide association study and genetic fine-mapping to identify the genetic cause of auto-flowering in cannabis. We then used gene expression analyses and transient transformation assays to characterize flowering time control. Herein, we identify a splice site mutation within circadian clock gene PSEUDO-RESPONSE REGULATOR 37 (CsPRR37) in auto-flowering cannabis. We show that CsPRR37 represses FT expression and its circadian oscillations transition to a less repressive state during SD as compared to long days (LD). We identify several key circadian clock genes whose expression is altered in auto-flowering cannabis, particularly under non-inductive LD. Research into the pervasiveness of this mutation and others affecting flowering time will help elucidate cannabis domestication history and advance cannabis breeding toward a more sustainable outdoor cultivation system.

Differentiation of hemp from marijuana using a qualitative decision-point assay

Marijuana and hemp are different cultivars of the species, Cannabis sativa. Trichomes within these genetically distinct forms result in different chemical constituents within the plant matrix. While drug-type cannabis yields higher total Δ9-THC concentrations, industrial or consumer-based hemp products are typically rich in cannabidiol or CBD-rich. Regulatory changes following the passage of agricultural legislation in the United States defines hemp as C. sativa containing no more than 0.3 % Δ9-THC on a dry weight basis. This threshold, which effectively differentiates legal hemp from illegal marijuana, presents a challenge to operational forensic laboratories. In this report we describe a decision-point assay to differentiate hemp from marijuana using a 1 % threshold. Methanolic extracts of C. sativa were analyzed using gas chromatography-mass spectrometry (GC–MS) using a deuterated analog (Δ9-THC-D3). Synchronous selected ion monitoring (SIM) and full scan acquisition was used for targeted analysis of Δ9-THC at the decision-point. Assay performance was evaluated in terms of limit of detection, linearity, carryover, selectivity, precision, accuracy and extract stability. Two analytical approaches are presented. Extraction efficiencies of Δ9-THC from plant matrix were 80–92 %, and decarboxylation of Δ9-tetrahydrocannabinolic acid (THCA) was 67 %. Interferences arising from the cyclization of CBD to produce Δ9-THC in the GC inlet were addressed using concentrations in excess of those typically encountered in plant samples. Accuracy was established across the specified range of the assay using known reference materials containing 0.12 to 10.1 % Δ9-THC. No false positive or negative results were identified (n = 140) using both analytical approaches.

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

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

Critical evaluation of analytical methodologies for the discrimination of hemp and drug-type cannabis

Classification of cannabis samples as hemp- or drug-type is an ever-increasing problem worldwide. Colorimetric tests and infrared spectroscopy were selected and critically compared among the available techniques. Cannabis samples with different THC and CBD concentration were analyzed by paper-based analytical devices (µPAD) using the 4-aminophenol colorimetric reaction, attenuated total reflectance mid infrared spectroscopy (ATR-MIR), and diffuse reflectance near infrared spectroscopy (DR-NIR). For the µPAD analysis, cannabis samples were extracted with methanol and later mixed with reagents to develop the colored complex. On the other hand, MIR and NIR spectra were obtained directly on raw samples, and partial least square discriminant analysis (PLS-DA) was used for cannabis classification. All the selected methodologies successfully discriminate between hemp-type and drug-type samples, being suitable to be used in-field. Additionally, assayed methodologies together a reference gas chromatography mass spectrometry (GC–MS) were evaluated and compared from a Green Analytical point of view using the AGREE criteria.

A forensic application of genetic markers for distinction between drug-type and fiber-type Cannabis sativa L.

Genetic markers can represent a valuable tool for forensic purposes in discriminating between fiber-type and drug-type cannabis. The aim of this research was to evaluate developed genetic markers for tetrahydrocannabinolic acid synthase (THCAS) when applied on certified hemp (14 varieties) and forensic casework samples of four chemotypes (40 seizures). Chemotype-associated PCR-based markers did not enable reliable selective amplification despite the difference in cannabinoid composition. In order to characterize forensic samples of unknown origin, THCAS sequencing was performed. The comparison of THCAS sequences, including additional accessions, indicated high genetic similarity of hemp varieties. Confiscated samples of intermediate, THC, CBD and CBG type were clearly separated from fiber-type accessions and assigned to drug-type cluster. Despite the unknown origin, their position on the tree support the notion that they are more related to drug-type accessions than to the fiber-type. However, no clear distinction between chemotypes was found. Furthermore, 26 amino acid substitutions were revealed in THCAS that clearly separate hemp varieties and neither of them cluster with any other tested sample.

Analysis of Morphological Traits in Herbarium of Historical Cannabis Specimens from Maghreb: Morphological Characteristics of Landrace Kif.

Background: Exploring the morphological traits of historical Cannabis populations from the Maghreb can provide invaluable information about the characteristics of the Maghreb Landrace Kif and contribute to the preservation of this vulnerable Cannabis resource. Aim: The purpose of the present study was to analyze the morphological characteristics of Cannabis specimens collected before worldwide introgressive hybridization and summarize the morphological traits of the Maghreb Landrace Kif. Discussion: Despite the limited number of specimens collected in the Maghreb, this study identified distinct types of Cannabis in the herbaria, including the Maghreb Landrace Kif, European hemp, and potentially East Asian hemp. By examining the morphological traits of kif specimens and reviewing the relevant literature, the study identifies the morphological traits that tend to characterize Maghreb Landrace Kif. Morphologically, Kif is different from drug-type Cannabis indica ("Sativa" and "Indica"). It resembles European hemp Cannabis sativa, but has female inflorescence characteristics that allow it to be distinguished from the latter, even when the growing conditions are optimal. These are the density of pistillate inflorescences (perigonal bract-to-leaf index), and the capitate stalked glandular trichomes cover density on the perigonal bracts. Conclusion: The characteristics of pistillate inflorescences identified in this study can be used to distinguish and select plants before phytochemical and genetic analysis, thus facilitating the identification of the Maghreb Landrace Kif.

Cultivar-dependent phenotypic and chemotypic responses of drug-type Cannabis sativa L. to polyploidization.

Cannabis sativa L. is a plant with a wide range of potential medicinal applications. In recent years, polyploidy has gained attention as a potential strategy for rapidly improving C. sativa, which, unlike other modern crops, has not yet benefitted from this established biotechnological application. Currently, no reports on high THCA and CBDA drug-type polyploid cultivars have been published. Moreover, it still needs to be clarified if different cultivars react similarly to polyploidization. For these reasons, we set out to evaluate and compare the phenotype and chemotype of three high Δ9-tetrahydrocannabinolic acid (THCA) and one high cannabidiolic acid (CBDA) drug-type cultivars in their diploid, triploid and tetraploid state through agronomic and metabolomic approaches. Our observations on plant morphology revealed a significant increase in plant height and leaf size with increasing ploidy levels in a cultivar-dependent manner. In contrast, cannabinoids were negatively affected by polyploidization, with the concentration of total cannabinoids, THCA, CBDA and cannabigerolic acid (CBGA) decreasing significantly in higher ploidy levels across all four cultivars. Headspace analysis of volatiles revealed that total volatile content decreased in triploids. On the other hand, tetraploids reacted differently depending on the cultivars. Two THCA dominant cultivars showed an increase in concentrations, while in the other two cultivars, concentrations decreased. Additionally, several rare compounds not present in diploids appeared in higher ploidy levels. Moreover, in one high THCA cultivar, a couple of elite tetraploid genotypes for cannabinoid and volatile production were identified, highlighting the role of cultivar and genotypic variability as an important factor in Cannabis sativa L. polyploids. Overall, our observations on plant morphology align with the giga phenotype observed in polyploids of other plant species. The decrease in cannabinoids and volatiles production in triploids have relevant implications regarding their commercial use. On the other hand, this study found that tetraploidization is a suitable approach to improve Cannabis sativa L. medicinal potential, although the response is cultivar and genotype-dependent. This work lays the ground for further improving, evaluating and harnessing Cannabis sativa L. chemical diversity by the breeding, biotechnological and pharmaceutical sectors.

Response of Medical Cannabis to Magnesium (Mg) Supply at the Vegetative Growth Phase.

Recent studies demonstrated a significant impact of some major macronutrients on function and production of medical cannabis plants, yet information on the effect of most nutrients, including Mg, is scarce. Magnesium is required for major physiological functions and metabolic processes in plants, and in the present study we studied the effects of five Mg treatments (2, 20, 35, 70, and 140 mg L-1 Mg), on plant development and function, and distribution of minerals in drug-type (medical) cannabis plants, at the vegetative growth phase. The plants were cultivated in pots under controlled environment conditions. The results demonstrate that plant development is optimal under Mg supply of 35-70 mg L-1 (ppm), and impaired under lower Mg input of 2-20 mg L-1. Two mg L-1 Mg resulted in visual deficiency symptoms, shorter plants, reduced photosynthesis rate, transpiration rate, photosynthetic pigments and stomatal conduction in young-mature leaves, and a 28% reduction of total plant biomass compared to the optimal supply of 35 mg L-1 Mg. The highest supply level of 140 mg L-1 Mg induced a small decrease in physiological function, which did not affect morphological development and biomass accumulation. The low-deficient Mg supply of 2 mg L-1 Mg stimulated Mg uptake and accumulation of N, P, K, Ca, Mn, and Zn in the plant. Increased Mg supply impaired uptake of Ca and K and their root-to-shoot translocation, demonstrating competitive cation inhibition. Mg-deficiency symptoms developed first in old leaves (at 2 mg L-1 Mg) and progressed towards young-mature leaves, demonstrating ability for Mg in-planta storage and remobilization. Mg toxicity symptoms appeared in old leaves from the bottom of the plants, under 140 mg L-1 Mg. Taken together, the findings suggest 35-70 mg L-1 Mg as the optimal concentration range for cannabis plant development and function at the vegetative growth phase.

Is Twelve Hours Really the Optimum Photoperiod for Promoting Flowering in Indoor-Grown Cultivars of Cannabis sativa?

Cannabis sativa ("cannabis" hereafter) is a valuable recent addition to Canada's economy with the legalization for recreational use in 2018. The vast majority of indoor cannabis cultivators use a 12-h light/12-h dark photoperiod to promote flowering. To test the hypothesis that robust flowering initiation responses can be promoted in indoor-grown cannabis cultivars under longer photoperiods, clones of ten drug-type cannabis cultivars were grown under six photoperiod treatments. All treatments were based on a standard 24-h day and included 12 h, 12.5 h, 13 h, 13.5 h, 14 h, and 15 h of light. The plants were grown in a growth chamber for 3 to 4 weeks, receiving an approximate light intensity of 360 µmol·m-2·s-1 from white LEDs. Flowering initiation, defined as the appearance of ≥3 pairs of stigmas at the apex of the primary shoot, occurred in all cultivars under all photoperiod treatments up to 14 h. Delays in flowering initiation time under 14 h vs. 12 h ranged from no delay to approximately 4 days, depending on the cultivar. Some cultivars also initiated flowering under 15 h, but floral tissues did not further develop beyond the initiation phase. Harvest metrics of some cultivars responded quadratically with increasing photoperiod, with ideal levels of key flowering parameters varying between 12 h and 13 h. These results suggest there is potential to increase yield in some indoor-grown cannabis cultivars by using longer than 12-h photoperiods during the flowering stage of production. This is attributed to the inherently higher daily light integrals. Indoor cannabis growers should investigate the photoperiod responses of their individual cultivars to determine the optimal photoperiod for producing floral biomass.

Characterization of Some "Hashish" Samples in the Egyptian Illicit Trafficking Market Using a Thermal Separation Probe and Gas Chromatography-Mass Spectrometry.

Drugs that are illegal have long been a part of Egyptian society. The most widely misused form of narcotic is marijuana, also known as "bango", and other cannabis-related products like "hashish". The chemical profile of some available "hashish" in the local Egyptian illegal market and its possible country of origin are investigated using a gas chromatography-mass spectrometry technique in conjunction with a thermal separation probe (TSP/GC/MS). The TSP/GC/MS method reveals the presence of 23 different terpenes, of which caryophylla-4(12),8(13)-dien-5α-ol, isoaromadendrene epoxide, caryophyllene, and alloaromadendrene oxide-(1) are detected in high relative proportions. Ten cannabinoid components are also detected. These are cannabiorcochromene (CBC-C1), tetrahydrocannabivarin (THCV), delta-8-tetrahydrocannabinol (delta-8-THC), exo-THC, cannabichromene, cannabidiol (CBD), cannabielsoin (CBE), dronabinol (delta-9-THC), cannabigerol (CBG), and cannabinol (CBN). Phenotypic index (THC % + CBN %)/CBD %) is measured for the test samples to identify both the nature of the samples (fiber- or drug-type cannabis) and the country of origin.

Sub-optimal nutrient regime coupled with Bacillus and Pseudomonas sp. inoculation influences trichome density and cannabinoid profiles in drug-type Cannabis sativa.

Cannabis sativa remains under heavy legal restriction around the globe that prevents extensive investigations into agricultural applications for improving its development. This work investigates the potential of specific plant growth-promoting rhizobacteria (PGPR) to improve Cannabis cannabinoid yield through increased trichome densities on floral organs, and to determine if sub-optimal environmental conditions would affect the outcomes of PGPR presence by altering plant development and cannabinoid profiles. Here, Pseudomonas sp. or Bacillus sp. were applied to the root system either separately or in a consortium to determine the effect of this bacterial treatment on the density of stalked glandular trichomes. Further, a low nutrient regime was applied for the first half of plant development to determine if an environmental stressor interacts with the effects of the microbial treatments on stalked trichome densities. Following 8 weeks of flower development, trichome density on calyces and bracts of inflorescences were determined using microscopy. Our findings unexpectedly indicate that recommended nutrient levels were linked to a decreasing trend in trichome densities with PGPR inoculations, but a low nutrient regime coupled with PGPR treatment increased them. Cannabinoid content is partially consistent with these results, in that a low nutrient regime increased the abundance of key cannabinoids compared to recommended regimes, with Bacillus sp. inoculation linked to the greatest number of significant changes between the two nutrient regimes. Overall, this work provides insight into how PGPR presence affects Cannabis stalked trichome development and cannabinoid profiles, and how environmental stressors can affect, and even enhance, trichome densities and influence major cannabinoid production, thereby pointing towards avenues for reducing the reliance on synthetic fertilizers during plant production without compromising yield.

Determination of 11 Cannabinoids in Cannabis sativa L. by Liquid Chromatography Quadrupole Time-of-Flight Mass Spectrometry (LC-Q-TOF-MS)

Eleven major cannabinoids from each subdivided tissue of drug-type and fiber-type cannabis plants were determined by means of a liquid chromatography quadrupole time-of-flight mass spectrometry (LC-Q-TOF-MS). The cannabinoids analyzed in this study were tetrahydrocannabinol acid (THCA), Δ9-tetrahydrocannabinol (Δ9-THC), cannabidiol acid (CBDA), cannabidiol (CBD), Δ8-tetrahydrocannabinol (Δ8-THC), cannabinol (CBN), cannabichromene (CBC), cannabidivarin (CBDV), cannabigerolic acid (CBGA), cannabigerol (CBG) and tetrahydrocannabivarin (THCV). As a result, THCA was detected in the bracts at 28.4 µg/mg, in the buds at 24.8 µg/mg, and in the leaves at 5.1 to 10.5 µg/mg in the drug-type cannabis plant. In addition, Δ9-THC, CBGA, CBN, CBG, CBC, and THCV were mainly detected in bracts, buds, and leaves. On the other hand, as for the fiber-type cannabis plant, CBDA was detected in the bracts at 27.5 µg/mg, in the buds at 10.6 µg/mg, and in the leaves at 1.5-3.3 µg/mg. In addition, Δ9-THCA, CBD, Δ9-THC, CBC, and CBG were mainly detected in bracts, buds, and leaves.

Effect of Explant Source on Phenotypic Changes of In Vitro Grown Cannabis Plantlets over Multiple Subcultures

Drug-type cannabis is often multiplied using micropropagation methods to produce genetically uniform and disease/insect-free crops. However, micropropagated plantlets often exhibit phenotypic variation, leading to culture decline over time. In cannabis, the source of these changes remains unknown, though several factors (e.g., explant's sources and prolonged in vitro culture) can result in such phenotypical variations. The study presented herein evaluates the effects of explant sources (i.e., nodal segments derived from the basal, near-basal, middle, and apical parts of the greenhouse-grown mother plant) over multiple subcultures (4 subcultures during 235 days) on multiplication parameters and leaf morphological traits of in vitro cannabis plantlets. While initial in vitro responses were similar among explants sourced from different regions of the plant, there were significant differences in performance over the course of multiple subcultures. Specifically, explant source and/or the number of subcultures significantly impacted plantlet height, number of nodes, and canopy surface area. The explants derived from the basal and near-basal parts of the plant resulted in the tallest shoots with the greatest number of nodes, while the explants derived from the middle and apical regions led to shorter shoots with fewer nodes. Moreover, the basal-derived explants produced cannabis plantlets with shorter but wider leaves which demonstrated the potential of such explants for in vitro rejuvenation practices with minimal culture decline. This study provides new evidence into the long-term impacts of explant source in cannabis micropropagation.

Cannabidiol in urine is not a proof of CBD consumption-lesson learned from urine sample analysis in routine caseworks.

Cannabidiol (CBD) has been gaining popularity in recent years. Knowing that CBD products can contain more tetrahydrocannabinol (THC) than expected, interpretation of cannabinoids concentration in urine can be tricky, especially when low amounts of THC and CBD are found. Moreover, interpretation can also be difficult due to interindividual variation in pharmacokinetics. The objective of this work was to take a critical look at the data from our daily practice as a toxicology laboratory. We have collected results obtained in a first batch of 1074 urine samples submitted to cannabinoids analysis, and results of cannabinoids content of a second batch of 719 seized materials. CBD was detected in 163 urine specimens (15%). Its concentration was higher than the limit of quantification of 5ng/mL in 108 samples only (10% of the sampling population). Most of CBD-positive samples were associated with a high THC-COOH concentration (> 500ng/mL in 63.8% of CBD-positive samples) suggesting only a few CBD consumers in our population. Cannabinoids composition of seized plant materials (drug type at first glance) revealed CBD in 110 of them (15% of the sampling population), with a concentration mostly below 1%. All of the resin samples were CBD positive, and contained more THC compared to flowers. We can conclude that urine samples from drug-type cannabis users contained a low amount of CBD, what was not described previously. These findings are useful for the interpretation of cannabinoids results in daily practice.