Effects of short-term environmental stresses on the onset of cannabinoid production in young immature flowers of industrial hemp (Cannabis sativa L.)

Backgrounds Cannabis sativa L. produces at least 120 cannabinoids. Although genetic variation is the main factor in cannabinoid production, the effects of short-term environmental stresses in the early flowering stage remains largely unknown. Methods To investigate the effects of short-term environmental stresses on the onset of cannabinoid production in young immature flowers, a hemp variety, Green-Thunder (5-8% CBD/mg of dry weight), was treated with mechanical damage, insect herbivory, extreme heat, or drought stress for 5-7 days during the first 2 weeks of flowering. Three hemp tissues, including flowers, leaves, and stems, were collected from hemp grown under these stress conditions at multiple time points during the first 2 weeks after transition to the short photoperiod and analyzed using high pressure liquid chromatography to quantify phytocannabinoids including cannabigerolic acid (CBGA), cannabigerol (CBG), cannabidiolic acid (CBDA), cannabidiol (CBD), Delta-tetrahydrocannabinolic acid (THCA), Delta-tetrahydrocannabinol (THC), and cannabinol (CBN). Results The 5 days of mechanical wounding did not affect the production of any of the cannabinoids during the initial stage of flowering. However, after 5 days of herbivore treatment, there was a significant difference in concentration between day 1 and day 6 of CBGA (control: 308 mu g/g; treatment - 24 mu g/g), CBG (control: 69 mu g/g; treatment: 52 mu g/g), and CBD (control: 755 mu g/g; treatment: 194 mu g/g) between the control and treatment plants. The 7 days of heat treatment at 45-50 C-o significantly reduced the production of CBGA during this observed window (control: 206 mu g/g; treatment: 182 mu g/g) and CBG (control: 21 mu g/g; treatment: - 112 mu g/g). Notably, the largest change was observed after 7 days of drought stress, when plants showed a 40% greater accumulation of CBG (control: 336 mu g/g; treatment: 622 mu g/g), and a significant decrease (70-80%) in CBD (control: 1182 mu g/g; treatment: 297 mu g/g) and THC amounts (control: 3927 mu g/g; treatment: 580 mu g/g). Conclusions Although this observation is limited in the early flowering stage, the common field stresses are adequate to induce changes in the cannabinoid profiles, particularly drought stress being the most impactful stress for hemp flower initiation with the altering the cannabinoid production by decreasing CBD and THC accumulation while increasing CBG by 40%.