In Vivo and In Vitro Chromosome Doubling of ‘I3’ Hemp

Abstract

Cannabis sativa L. is a diploid (2x) herbaceous plant that provides a wide variety of products such as essential oils, fiber, and medicine. Hemp was defined in the 2018 Farm Bill as a Cannabis plant with a delta-9 tetrahydrocannabinol concentration of not more than 0.3% on a dry-weight basis. Polyploidy is frequently used in plant breeding to manipulate vigor, reproductive fertility, and biochemistry. By inducing polyploidy/chromosome doubling, we may increase the compounds of interest, principally CBD (cannabidiol), produced by hemp. The purpose of this experiment was to evaluate the efficacy of different treatments of colchicine and oryzalin applied in vivo and in vitro to induce polyploidy in ‘I3’ hemp. After treating vegetative cuttings with colchicine or oryzalin, we had a 31% survival rate. Of the 85 survivors, we recovered two tetraploids: one from the 12-h 0.05% colchicine treatment group and the other from the 12-h 0.2% colchicine treatment group. For the in vitro portion of the experiment, the 12-h 50-µM oryzalin treatment yielded one tetraploid and the 36 h 50 µM oryzalin treatment yielded one cytochimera (mixoploid). The relative efficiency of some treatments showed potential for a simple method to induce tetraploids in clonal hemp for breeding.