In vitro flowering and micropropagation are useful for plant breeding programs and commercial production of important ornamental plants. In vitro conditions including media components, kind, concentration and ratio of plant growth regulators and culture conditions significantly affect in vitro flowering and micropropagation. There is no any report dealing with the in vitro flowering of Lisianthus (Eustoma grandiflorum). Here, a protocol was developed for flowering and high frequency in vitro micropropagation of E. grandiflorum, an ornamental plant. Micropropagation is an effective tools for propagation of ornamental plants in large scale. The aim of the present study was to evaluate the effect of different concentrations of NAA and BA on micropropagation and flowering of Lisianthus, in vitro. Used culture medium was MS enriched with 0, 0.1, 0.2 and 2 mg L^-1 of NAA and BA. In establishment process of explants, the most shoot length (2.07 cm per plant) was obtained on medium supplemented with 0.1 mg L^-1 BA (without NAA). Maximum shoot number (5.80 per plant) was produced in medium containing 0.1 mg L^-1 BA along with 0.2 mg L^-1 NAA. Bud explants in culture media containing 0.2 mg L^-1 NAA (without BA) and 0.1 mg L^-1 NAA along with 2 mg L^-1 BA produced maximum node number (3.20 per plant). The largest number of root (14.53 per plant) and root length (3.87 cm per plant) were produced on 0.2 mg L^-1 NAA without BA, also 0.2 mg L^-1 BA plus 0.2 mg L^-1 NAA and 0.2 mg L^-1 BA without NAA. Explants produced flower on medium containing 0.1 mg L^-1 BA along with 0.1 mg L^-1 NAA without transition of callus formation. Flower was produced from callus in medium containing 0.1 mg L^-1 BA along with 2 mg L^-1 NAA. Regenerated plants showed 98% survival in greenhouse during acclimatization. Acclimatized plants were morphologically similar to the mother plants.

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