The standard practice to initiate flowering in medicinal cannabis involves reducing the photoperiod from a long-day period to an equal duration cycle of 12 h light (12L)/12 h dark (12D). This method reflects the short-day flowering dependence of many cannabis varieties but may not be optimal for all. We sought to identify the effect of nine different flowering photoperiod treatments on the biomass yield and cannabinoid concentration of three medicinal cannabis varieties. The first, “Cannatonic”, was a high cannabidiol (CBD)-accumulating line, whereas the other two, “Northern Lights” and “Hindu Kush”, were high Δ9-tetrahydrocannabinol (THC) accumulators. The nine treatments tested, following 18 days under 18 h light/6 h dark following cloning and propagation included a standard 12L:12D period, a shortened period of 10L:14D, and a lengthened period of 14L:10D. The other six treatments started in one of the aforementioned and then 28 days later (mid-way through flowering) were switched to one of the other treatments, thus causing either an increase of 2 or 4 h, or a decrease of 2 or 4 h. Measured parameters included the timing of reproductive development; the dry weight flower yield; and the % dry weight of the main target cannabinoids, CBD and THC, from which the total g cannabinoid per plant was calculated. Flower biomass yields were highest for all lines when treatments started with 14L:10D; however, in the two THC lines, a static 14L:10D photoperiod caused a significant decline in THC concentration. Conversely, in Cannatonic, all treatments starting with 14L:10D led to a significant increase in the CBD concentration, which led to a 50–100% increase in total CBD yield. The results show that the assumption that a 12L:12D photoperiod is optimal for all lines is incorrect as, in some lines, yields can be greatly increased by a lengthened light period during flowering.
With an increasing appreciation for the unique pharmacological properties associated with distinct, individual cannabinoids of Cannabis sativa, there is demand for accurate and reliable quantification for a growing number of them. Although recent methods are based on highly selective chromatography-mass spectrometry technology, most are limited to a few cannabinoids, while relying on unnecessarily sophisticated and expensive ultra-high performance liquid chromatography and tandem mass spectrometry. Here we report an optimised, simple extraction method followed by a reliable and simple high performance liquid chromatography method for separation. The detection is performed using a time-of-flight mass spectrometer that is available in most natural products research laboratories. Due to the simplicity of instrumentation, and the robustness resulting from a high resolution in the chromatography of isobaric cannabinoids, the method is well suited for routine phytocannabinoid analysis for a range of applications. The method was validated in terms of detection and quantification limits, repeatability, and recoveries for a total of 17 cannabinoids: detection limits were in the range 11–520 pg when using a 1 µL sample injection volume, and the recovery percentages ranged from 85% to 108%. The validated method was subsequently applied to determine cannabinoid composition in the inflorescences of several medicinal Cannabis sativa varieties.
The standard two-step methods for grafting horticultural crops involve cultivating the rootstock for a period of time and then connecting the scion. Medicinal Cannabis differs from most annual horticultural crops because it is usually clonally propagated from cuttings. We developed a grafting methodology specifically for medicinal Cannabis, involving a single step, in which a freshly cut scion is grafted to a freshly cut donor stem that will become the rootstock. This study also aimed to uncover a potential role for roots in influencing cannabinoid content. Two varieties with desirable attributes but cultivation limitations were selected to act as scions. The first, “CBD1” was a high CBDA accumulating variety with low biomass yield, and the second, “THC2”, was a high yielding, high THCA accumulating line with inconsistent root development during cloning. Two candidate rootstocks, “THC9r” and “THC8r”, were identified; both were high THCA, low CBDA varieties. Biomass yields in the THC2 scions grafted to THC9 rootstocks (THC9r_2s) were 20% higher than in the non-grafted THC2 plants. In CBD1 grafted plants, the concentrations of CBDA and some minor cannabinoids were significantly different to non-grafted CBD1, but biomass yields were lower. There was a trend towards a higher concentration of THCA in THC9r_2s plants, and when combined with the increased biomass, yield of THCA was increased from 8 g Plant−1 to 13 g Plant−1. Our results present a new grafting method for medicinal Cannabis that improved yield in THC2 and required no additional cultivation time.
Chickpea is the second-most-cultivated legume globally, with India and Australia being the two largest producers. In both of these locations, the crop is sown on residual summer soil moisture and left to grow on progressively depleting water content, finally maturing under terminal drought conditions. The metabolic profile of plants is commonly, correlatively associated with performance or stress responses, e.g., the accumulation of osmoprotective metabolites during cold stress. In animals and humans, metabolites are also prognostically used to predict the likelihood of an event (usually a disease) before it occurs, e.g., blood cholesterol and heart disease. We sought to discover metabolic biomarkers in chickpea that could be used to predict grain yield traits under terminal drought, from the leaf tissue of young, watered, healthy plants. The metabolic profile (GC-MS and enzyme assays) of field-grown chickpea leaves was analysed over two growing seasons, and then predictive modelling was applied to associate the most strongly correlated metabolites with the final seed number plant−1. Pinitol (negatively), sucrose (negatively) and GABA (positively) were significantly correlated with seed number in both years of study. The feature selection algorithm of the model selected a larger range of metabolites including carbohydrates, sugar alcohols and GABA. The correlation between the predicted seed number and actual seed number was R2 adj = 0.62, demonstrating that the metabolic profile could be used to predict a complex trait with a high degree of accuracy. A previously unknown association between D-pinitol and hundred-kernel weight was also discovered and may provide a single metabolic marker with which to predict large seeded chickpea varieties from new crosses. The use of metabolic biomarkers could be used by breeders to identify superior-performing genotypes before maturity is reached.
Far-red (FR) light elicits two distinct processes in plants. First, a shade avoidance response which is triggered when the ratio of red to FR (R: FR) declines. Second, it interacts synergistically with higher frequency wavelengths of light (e.g. red or white) which improves the efficiency of photosynthesis. We investigated whether we could harness these phenomena in medicinal Cannabis to improve yields so that the duration of the photoperiod could be reduced to 10 h (“10L”) whilst returning similar or improved yields compared to a 12 h photoperiod (“12L”). The THC concentrations were elevated in both high THC varieties by the different FR treatments. In Hindu Kush the concentration of THC was elevated by the addition of 4 h of total FR (“10L_2_2D”), and in Northern Lights total cannabinoid yields were increased by nearly 70% over the 12 L control (0.43 versus 0.25 g Plant− 1) by the addition of 2 h of FR in darkness after 10 h of light (“10L_2D”). Our results show a strong yield and quality advantage in high THC lines treated with end-of-day FR treatments. Furthermore, a lighting schedule of 10L_2D instead of 12 L would result in a saving of 5.5% in power usage and resultant emissions.