Basal media optimization for the micropropagation and callogenesis of Cannabis sativa L.

Micropropagation of Cannabis sativa is an emerging area for germplasm storage and large-scale production of clean plants. Existing protocols use a limited number of genotypes and are often not reproducible. Previous studies reported MS + 0.5 M TDZ to be optimal for Cannabis nodal micropropagation, yet our preliminary studies using nodal explants suggested this media may not be optimal. It resulted in excessive callus formation, hyperhydricity, low multiplication rates, and high mortality rates. Following an initial screen of four commonly used basal salt mixtures (MS, B5, BABI, and DKW), we determined that DKW produced the healthiest plants. In a second experiment, the multiplication rate and canopy area of explants grown on MS + 0.5 M TDZ and DKW + 0.5 M TDZ were compared using five drug-type cultivars to determine if the preference for DKW was genotype-dependent. Four cultivars had significantly higher multiplication rates on DKW + 0.5 M TDZ with the combined average being 1.5x higher than explants grown on MS + 0.5 M TDZ. The canopy area was also significantly larger on DKW + 0.5 M TDZ for four cultivars with the combined average being twice as large as the explants grown on MS + 0.5 M TDZ. In the third experiment, callogenesis was compared using a range of 2,4-D concentrations (0-30 M) on both MS and DKW and similarly, callus growth was superior on DKW. This study presents the largest comparison of basal salt compositions on the micropropagation of five commercially grown Cannabis cultivars to date.