PLANT CELL BIOTECHNOLOGY AND MOLECULAR BIOLOGY

The present study was conducted to establish a protocol for direct and indirect regeneration from various explants of Indigofera tinctoria and Indigofera enneaphylla as well as to develop a well proliferating callus/suspension culture containing well organized group of calli that can be used for isolation of secondary metabolites. Shoot tip, nodal and leaf disc explants were grown on full strength modified MS medium with NAA and BAP. Shoot tip explants of Indigofera tinctoria induced multiple shoots through axillary budding in MS medium containing 0.5 mg/l each of NAA and BAP. Enhancing the concentration of BAP to 1.0 mg/l induced a well proliferating callus from shoot tips. The callus regenerated to produce multiple shoots. Similarly, in the case of Indigofera enneaphylla also multiple shoots were obtained from cultured shoot tips through axillary budding in MS medium with 1.0 mg/l each of NAA and BAP. On the contrary, shoot tips incubated on 0.5 mg/l NAA and 1.0 mg/l BAP initiated well proliferating callus, which on further incubation regenerated multiple shoots. Nodal explants of Indigofera enneaphylla induced well proliferating callus in MS medium supplemented with 0.5 mg/l NAA and 1.0 mg/l BAP. Regeneration of shoots was obtained from the callus. Shoot formation by axillary budding in medium containing 0.5 mg/l each of NAA and BAP from the nodal explants was also reported. Leaf disc explants of both species developed a well proliferating callus in MS medium supplemented with 0.5 mg/l NAA and1mg/l BAP. The callus so obtained on subculture to suspension containing 1/2 MS/SH with 1/2 sucrose supplemented with 0.5 mg/l NAA and 0.5 mg/l BAP produced organization of callus into globular clumps. Such organized callus is conducive for secondary metabolite production. Regeneration of multiple shoots from leaf callus of I. tinctoriaand I. enneaphylla is also reported in this study.