Genetic and Metabolic Studies of Cannabinoids in Standardized Medicinal Cannabis sativa

In this research we investigated the biosynthesis and accumulation of cannabinoids during the growth phases of Cannabis sativa leaves and flowers. Flowers from standardized indoor breeding were analyzed for transcription and expression of identified genes [1–5] from the cannabinoid pathway and the accumulation of the cannabinoid metabolites [6]. The correlation between the various measurements should give more information on the regulation of the cannabinoid production process within the plant. Plant samples were taken randomly during standardized cultivation. Every week, for eight weeks in a row, three plants were sampled, and materials were treated for analysis by QRT-PCR, HPLC, and 2D-electrophoresis. With QRT-PCR the transcription of CBDA-(BAF65035), THCA-(BAE48253) and olivetol synthase (BAG14339) genes were quantified against cloned genes. 2D-electrophoresis was used to detect any specific protein expression during the cultivation period. From this ongoing study, we have indicated that the amount of THCA in the leaves stays in certain ranges throughout the sampling period and is not dependant on the vegetative or flowering status of the plant. In contrast, the content of THCA in the flowers is depending on the growth period, which is in line with previously reported data on the correlation of trichoma and cannabinoids. The information obtained from this study is used as a profound basis for further genetic and metabolic analysis. References: [1] Kim JS, et al. (2006) Biotechnol Lett. 28(13): 999–1006. [2] Sirikantaramas S, et al. (2005) Plant Cell Physiol. 46(9): 1578–1582. [3] Sirikantaramas S, et al. (2004) J Biol Chem. 279(38): 39767–39774. [4] Morimoto S. et al. (1998) Phytochemistry, 49(6): 1525–1529. [5] Taura F. et al. (1996) J Biol Chem. 271(29): 17411–17416. [6] Fellermeier M, et al. (2001) Eur J Biochem. 268(6): 1596–1604.