IN VITRO CALLUS INDUCTION AND PLANT REGENERATION IN BRYONIA LACINIOSA FROM LEAF

Abstract

 This study investigates effective sterilization methods for the in vitro culture of Bryonia laciniosa (Linn.) in 2022-23, revealing optimal disinfection with 0.2 per cent sodium hypochlorite followed by fungicide (Sofia) treatment. Although 0.3 per cent mercuric chloride yielded high explant survival, it elevated culture mortality, favoring sodium hypochlorite for successful disinfection. An efficient protocol for callusing and regeneration via direct and indirect means was established using leaf segments. The different forms of callus such as compact, nodular and green callus were produced only multiple shoots. Better quality of pure callus was obtained in the medium fortified with MS nutrients, 3.0 mg/L 2,4-D. The induced callus produced multiple shoots on the same mother medium after 3-4 weeks of culture. The induced shoots were also elongated in the same mother medium without the addition of any other plant growth regulator. 7 shoots  were regenerated in leaf explants in MS medium with KIN (4 mg/L) and IAA (1 mg/L).Elongated shoots took roots supplemented with 4.0 mg/L KIN and 1.0 mg/L IBA. The regenerated plants were successfully hardened into pots after properacclimatization. These findings underscore the potential of explants for callus induction, morphological analysis, and indirect plantlet regeneration, emphasizing the importance of in vitro techniques in preserving endangered medicinal plants.