For the development of an efficient callus initiation and direct organogenesis protocol in Scoparia dulcis Linn., an important medicinal plant, nodal segments of field grown plants were aseptically cultured on agar solidified MS medium supplemented with different concentrations and combinations of three PGRs, namely IAA, NAA and BA. The explants produced a green or light green compact callus on MS medium fortified with 0.5-2.0 mg/L BA in combination with 0.5-1.0 mg/l IAA or NAA. The maximum amount of callus was produced on MS with 1.5 mg/l BAP + 0.5 mg/l NAA in the case of explants. These callus tissues underwent differentiation when grown on a range of PGRs (BAP, IAA and NAA) supplemented media. The maximum number of callus was developed in medium containing 1.5 mg/l BAP + 0.5 mg/l NAA. Direct organogenesis underwent rapid elongation on elongation media and maximum elongation took place on MS with 1.5 mg/l BAP + 0.5 mg/l IAA. Antimicrobial activity of ethanol extracts of S. dulcis was also determined by using various bacterial strains. Ethanolic extracts of S. dulcis showed moderate activity against various organisms.