Investigation of factors in improving Agrobacterium-mediated gene transfer in Ruellia tuberosa L. and evaluation of α-glucosidase inhibitory activity in established hairy roots

Ruellia tuberosa (family Acanthaceae) is widely known in traditional medicine Asian countries for the treatment of diabetes and other diseases. Its roots were demonstrated to possess a hypoglycemic ability diabetic animal models. In this study, an original induced procedure was investigated establish hairy root (HR) from R. tuberosa. With aim increasing transformation rate, some factors (acetosyringone (AS) dosage, type explant, age, infection time, bacterial density, co-cultivation duration) individually examined. As result, improved implemented: ten-day-old vitro cotyledon explants injured then immersed suspension (OD600 nm = 0.4) added 200 µM AS during 10 min. The infected co-cultivated 4 days Murashige & Skoog (MS) medium before transferring containing cefotaxime elimination. After thirty culture, revealed synergistic effect by enhancing rooting rate number secondary per explant up 4.4- 8.0-fold, respectively, comparison with procedure. HR cultured liquid MS achieved highest biomass production at late exponential growth phase (3rd week). ethanol extract also higher 2.0-fold α-glucosidase inhibitory activity than that natural root. conclusion, inducing may offer effective reliable substitute utilization herbal plant.