Cannabis sativa is an industrial crop producing more economic value than the top five crops in California combined. The FDA has recently approved two cannabinoids for use to treat and alleviate symptoms for a wide range of diseases, including nausea in cancer patients undergoing chemotherapy, while studies are examining their use for reduction of anxiety, inflammation, and pain relief. The female plant produces cannabinoids, and two factors known to reduce cannabinoid production are pollination and stress. Sex determination in cannabis is therefore an important economic factor. DNA methylation is an important epigenetic mechanism that regulates many cellular processes, including sex determination and seed production.
Enzymatic Methyl-seq (EM-seq TM) is a novel enzymatic method used to detect 5mC and 5hmC. Here, we used EM-seq to better understand sex determination and the hermaphroditic process in cannabis. To gain insight into methylation based regulation of the hermaphroditic process, EM-seq libraries were prepared using genomic DNA from female flowers, female seeded flowers and male flowers. EM-seq successfully identified 5mCs and 5hmCs across the cannabis genome. Additionally, RNA-Seq data was generated to establish functionality between DNA methylation and the cannabis transcriptome. EM-seq enabled the generation of the first cannabis methylome and opens up the possibility to discover critical and exciting regulatory functions for the hermaphroditic process and cannabinoid pathway regulation.