As the industrial hemp (Cannabis sativa L.) market grows, there is a need for methods to clonally propagate parental breeding stock and new cultivars. Information is lacking on vegetative cutting propagation of hemp. We evaluated how propagation environment (intermittent mist vs. subirrigation under a humidity dome), indole-3-butyric acid (IBA) formulation (talc rooting powder vs. IBA in solution), and IBA concentration (0, 3000, or 8000 ppm) affected stem cuttings from ‘I3’, a cannabinoid-free cultivar of industrial hemp. Under mist or domes, rooting quality and percent declined at 8000 ppm IBA. Root and shoot quality and rooting percentage also were reduced in 3000 ppm IBA in solution treatment compared with talc. Our data show that for the cultivar tested, cuttings rooted at the highest percentage and produced the highest-quality roots and shoots with either no hormone or 3000 ppm talc powder. These treatments did equally well under humidity domes or intermittent mist.

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.