Low Caffeine Concentrations Induce Callus and Direct Organogenesis in Tissue Cultures of Ornithogalum dubium

Caffeine is a nitrogenous base that naturally occurs in coffee (Cafea arabica), tea (Thea sinensis), and cocoa (Theobroma cacao). Chemically, caffeine is 1,3,5-trimethylxanthine, a purine analogue. Due to significant human consumption, caffeine effects have been widely studied. Being a natural xanthine derivative, the key degradative enzyme is xanthine oxidase, converting caffeine into 1-methyluric acid. Ecologically, caffeine is believed to act as a repellent molecule against insect feeding behavior. Caffeine's chemical similarity to purines and plant hormones motivated this study, establishing the potential for cellular de-differentiation and re-differentiation. For this, a highly hormone-responsive plant species, Ornithogalum dubium, was used. As caffeine has been shown to induce endoreplication, the potential for new germlines in O. dubium is attractive. Using tissue culture, a range of caffeine concentrations were used (0.0125 mg/L to 2.0 mg/L) without additional hormones. A significant difference (p > 0.05) was observed for intermediate concentrations of 0.0125, 0.025, and 0.05 mg/L when compared to the control (no hormones). The highest rates of callus induction were obtained at a concentration of 0.025 mg/mL. Higher concentrations were phytotoxic (1.0 mg/L or greater). To conclude, caffeine-regenerated plants were not dissimilar to those obtained from canonical hormones.