Double haploid development in drug-type Cannabis sativa L. through microspore indirect de-novo plant regeneration

Double haploid technology (DH) is an essential tool in plant breeding, enabling the rapid production of homozygous lines. However, Cannabis sativa L. has been categorized as recalcitrant to DH induction. In this study, we evaluated the potential to generate DH C. sativa plants via anther culture and indirect de-novo organogenesis. We examined a THCA-dominant cultivar with a callus induction success of 29.48%. Mixoploidy in the callus indicated spontaneous genome doubling. This is the first report documenting the successful induction of DH C. sativa plants through de-novo indirect organogenesis. These findings have profound implications for the C. sativa breeding sector by potentially improving efficiency of genome editing and hybrid development in this economically significant species.