Time Dependent Metabolomics and Transcriptional Analysis of Cannabinoid Biosynthesis in Cannabis sativa var. Bedrobinol and Bediol Grown under Standardized Condition and with Genetic Homogeneity

Abstract

Cannabis sativa L. is mostly grown under non standardized conditions resulting in high fluctuations of cannabinoid contents. The exact measure of these metabolites is hampered as most of the research has been conducted on seized, extracted or non-standardized cultivated C. sativa L. material. In this report legal standardized cultivation of two C. sativa L. chemotypes is investigated by HPLC-UV, qPCR and 1 H-NMR, respectively during two cultivation phases of four (4) weeks each. Based on HPLC the cannabinoid spectrum over 4 week cultivation period is monitored under specific light conditions and in correlation with the expression profile of thca and cbda genes in both chemotypes. Cannabinoid accumulation is observed specifically after shortening of the light period. Gene transcript based chemotyping was only possible comparing the thca and cbda transcription level ratio at the middle of the cultivation period, while at the end differences in gene transcription ratio diminished. 1 H-NMR results indicate a clear difference between both chemotypes and cultivation phases by applying pattern recognition software. The differences detected by using simple and non-destructive 1 H-NMR were used to build a model, which can be applied to chemotyping and cultivation phase separation using selective 1 H-NMR signals.