A simple and efficient micropropagation system through direct shoot regeneration has been developed through leaf disc culture of Sansevieria cylindrica. BA (10 µM) was most effective in inducing bud break and growth, as well as in initiating multiple shoot formation at the rate of 10 microshoots per leaf disc explant, after 4 weeks of culture. The highest shoot regeneration frequency (80%) and the highest number of shoots per explant (13.5 ± 0.22) was obtained in MS medium supplemented with 10 µM BA and 0.1 µM NAA. The regenerating tissues after harvesting the shoots were subcultured on MS medium supplemented with reduced concentration of BA (5 µM), where a high frequency multiplication rate was established. The regeneration potential was unaffected up to fourth subculture and afterwards it decreased drastically. With 5 µM IBA, efficient rooting was achieved for 80% of the microshoots. The regenerated shoots displayed a unique characteristic of flattened leaf blade in the beginning as compared to cylindrical blade in parental plants but acquired normal leaf morphology after 18 months of acclimatization. Plantlets were hardened in a controlled plant growth chamber, prior to ex vitro transfer, and the survival rate was 90%.