From dawn 'til dusk: daytime progression regulates primary and secondary metabolism in Cannabis glandular trichomes.

Cannabis sativa L. glandular trichomes synthesize large amounts of secondary metabolites, predominantly cannabinoids and terpenoids. The associated demand for carbon and energy makes glandular trichomes strong sink tissues with indications that their secondary metabolism is coupled to the availability of photoassimilates. Many metabolites show diurnal patterns of flux, but it is unknown whether cannabinoids and terpenoids are regulated by time of day. We quantified cannabinoids, terpenoids, and the glandular trichome proteome over a 12 h light period in flowers of 'Hindu Kush', a high-tetrahydrocannabinol cultivar. Major cannabinoids changed significantly over the course of the day, resulting in an increase in total measured cannabinoids. Major terpenoids also changed, with sesquiterpenes generally decreasing with day progression. While monoterpenes generally did not decrease, the second most abundant, α-pinene, increased. The glandular trichome proteome changed the most within the first 6 h of the day, and analysis of differentially abundant proteins indicated up-regulation of primary metabolism. Surprisingly, key cannabinoid biosynthetic enzymes decreased with daytime progression despite increases in cannabinoid content, which indicates that daytime increases of photoassimilates are the main driver of cannabinoid regulation. This first reporting of variability of cannabinoid and terpenoid biosynthesis over the course of the day has implications for Cannabis research and production.