In vitro regeneration and transformation of Coleus forskohlii with glucanase-chitinase gene

Coleus forskohlii Briq. is a threatened medicinal plant was transformed with glucanase-chitinase gene along with npt-II gene. The transformants were obtained on MSO supplemented with 2 mg/l or 3 mg/l BAP and 0.2 mg/l 2,4-D for callus induction using leaf as explants. Multiple shoot production from the leaf-derived transformed callus of Coleus has been standardized using the various concentration of NAA with 2 mg/l BAP. The combination of 2 mg/l BAP and 0.5 mg/l NAA was found to be best for multiple shoot production. Rooting of the transformed shoots was found to be best on MS medium supplemented with 0.7 mg/l NAA. The presence of npt-II genes in the leaves of putative transformants was confirmed by PCR analysis. SDS-PAGE analysis and glucanase assay revealed the over expression of glucanase enzyme in the transgenic plants. The transgenic Coleus plants survived the infection caused by Fusarium chlamydosporium due to the accumulation of glucanase-chitinase enzyme, whereas control plants were susceptible to the disease.