Numerous shoots were directly regenerated from the leaf explants of Lysionotus pauciflorus on the MS medium containing 0.5–2 �M NAA with or without 1 �M BA. The calli were induced from the leaves on MS medium supplemented with 2 �M 2, 4-D. The calli proliferated about four times in fresh weight in the liquid medium of the same composition as the callus induction medium after 4 weeks of culture on a rotary shaker at 100 rpm. Shoots were induced from these calli on the regeneration medium amended with 32 �M BA or 0.5 �M zeatin. Regenerated shoots rooted easily on � MS medium without any plant growth regulators. Most of the regenerants from callus were diploid, whereas eight of 66 acclimatized plantlets were tetraploid determined by flow cytometric analysis. Furthermore, flow cytometric analysis of calli also revealed tetraploidy.