IN-VITRO SEED GERMINATION AND EFFECT OF TDZ AND AgNO3 ON HIGH FRE-QUENCY SHOOT REGENERATION FROM RUELLIA TUBEROSA L., USING COTYLE-DANARY NODE EXPLANTS

Ruellia tuberosa, commonly called Minnie root, has plenty of applications in herbal medicine due to the presence of several bioactive compounds. A simple, efficient, and reproducible in-vitro plant regeneration protocol was developed for Ruellia tuberosa. The cotyledonary nodal explants were excised from seedlings were inoculated on Murashige and Skoog (MS) medium supplemented with various concentrations of BAP and NAA, respectively. The highest numbers of 9.86 ± 0.15 shoots/explant were obtained in MS medium augmented with BAP (2.0 mg L^-1) and NAA (1.0 mg L^-1). The micro shoots induced per explant significantly increased with the supplementation of BAP to auxin-containing medium. The results showed that maximum percentage of multiple micro shoots 89%, with a number (13.80 ± 0.26) shoots / explant was found on MS medium supplemented with BAP (2.0 mg L^-1) and NAA (1.5 mg L^-1). Further, the highest frequency of micro shoots was produced on MS medium supplemented with TDZ (0.3 mg L^-1) and AgNO₃ (0.4 mg L^-1) for large-scale production. The highest frequency of root induction (96%) was recorded on a half-strength MS medium fortified with 1.5 mg L^-1 IBA. The well-rooted plantlets were transferred into the plastic cups containing peat and soil (2:1) and successively established in the greenhouse. In conclusion, our study demonstrates efficient in-vitro regeneration, and in the future, this protocol will be a valuable tool to enhance secondary metabolite production in Ruellia tuberosa.