Induction of somatic embryogenesis in leaf and root explants of Digitalis lanata Ehrh.: Direct and indirect method
Bhushan P. Bhusare, C. K. John, Vijay Bhatt, Tukaram D. Nikam
Abstract
Somatic embryogenesis was reported via direct and indirect method in cardio-tonic plant Digitalis lanata Ehrh. MS basal medium supplemented with cytokinins BAP, Kinetin (Kin) and TDZ, either alone or in combinations with auxins IAA, 2,4-D and IBA,- at different concentrations (00–3.0 mg/l) were studied. The best direct induction of somatic embryogenesis was observed in leaf (100% response with 11.3 SEs/explant) and root (85.7% response with 7.2 SEs/explant) explants on MS medium fortified with combination of Kin and IBA (2.0 mg/l each). Among the combinations tried, 2,4-D with Kin both at 1.0 mg/l was superior for the production of embryogenic calli and indirect induction of somatic embryos (95.2% with 9.8 SEs/explant). Incorporation of PEG (10 mg/l) in MS medium was beneficial for differentiation (13.0 ± 1.5 globular, 16.4 ± 1.4 heart and 19.8 ± 0.7 cotyledonary) and maturation of somatic embryos. SEM study confirmed the origin of somatic embryos directly from explants, and different developmental stages. The somatic embryo derived plantlets were hardened and successfully established in the field. As the direct and indirect somatic embryogenesis protocols are played significant role in genetic transformation, thereof thus the protocol reported here having importance in genetic improvement of a cardio-tonic herb D. lanata using recombinant DNA technology.