The capability of synthetic cytokinin mimics for in vitro regeneration of plants

Among plant hormones, cytokinins have been extensively studied for over a century due to their central roles in regulating plant development, physiology, and environmental responses. In this study, we chemically synthesized several Cytokinin-Active ingreDients (CADs) and evaluated their effects on the in vitro regeneration of Sansevieria trifasciata and seedling growth of Nicotiana tabacum , comparing their activities with established cytokinins, kinetin and zeatin. Our results show that CADs promote cell division and shoot proliferation in S. trifasciata , with the highest shoot numbers per explant observed at CAD-A (0.9 mg L -1 ), cycle-iP (0.1 mg L -1 ), and zeatin (0.3 mg L -1 ), all significantly different from kinetin (0.3 mg L -1 )—the previously suggested cytokinin for in vitro shoot proliferation in this plant. CAD-A and CAD-B at 0.9 mg L -1 increased leaf number by approximately 4.5, significantly outperforming kinetin at 0.3 mg L -1 . In tobacco seedlings, CAD-B at 0.3 mg L -1 significantly enhanced root number (7.3) comparable to kinetin (6.7) at the same concentration. Overall seedling height increased more than two-fold under both CAD-B and kinetin treatments compared to control plants. The bioactivity of CADs under in vitro conditions closely parallels that of kinetin, supporting their potential as effective cytokinin mimetics. Collectively, these results indicate that synthesized CADs can serve as potent alternatives to natural cytokinins for plant tissue culture and developmental studies, with promising applications in propagation and regeneration protocols. • Chemically synthesized cytokinin mimetics (CADs) effectively promote shoot and root development in snak plant and Nicotiana under in vitro conditions. • CAD compounds induce comparable or superior plant growth responses to traditional cytokinins like kinetin and zeatin. • The study supports the potential of synthetic cytokinin analogs as versatile alternatives for plant tissue culture and regeneration protocols. • Structural differences in cytokinin mimetics influence bioactivity, highlighting opportunities for tailored plant growth regulation strategies.