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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.
A protocol was developed for high frequency and low cost of in vitro shoot proliferation and ex vitro rooting of Eustoma grandiflorum (Gentianaceae) on solid medium. Shoot tips as explants were cultured on Murashige and Skoog (MS) medium enriched with different concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D) (0.00, 0.01, 0.10 and 1.00 mg l–1) and 6-benzylaminopurine (BAP) (0.00, 0.50, 2.00 and 5.00 mg l–1). Three culture media systems (solid, liquid and double-phase) were applied. None of the explants cultured on liquid and double-phase media resulted in live plant production. Maximum axillary shoot number (54.45) was recorded in the plantlets treated with 0.10 mg l–1 2,4-D in combination with 5.00 mg l–1 BAP. Treatment of 0.01 mg l–1 2,4-D along with 0.50 mg l–1 BAP produced maximum node number and internode length. Some shoots produced on medium containing plant growth regulators (PGRs) were rooted in soil. The largest number (5.50/plantlet) and longest length of root (7.75 cm/plantlet) were obtained in ex vitro condition on the base of shoots produced in culture medium enriched with 0.10 mg l–1 2,4-D along with 0.50 mg l–1 BAP. The combination of 1.00 mg l–1 2,4-D and 0.50 mg l–1 BAP was found to be the most suitable PGRs for obtaining the highest callus weight. The most fresh weight was calculated from plantlets grown on the medium containing 0.10 mg l–1 2,4-D along with 5.00 mg l–1 BAP. Maximum dry weight was obtained in free-PGRs medium. About 90% of the rooted plantlets were established successfully in cultivation beds. Acclimatized plants were morphologically similar to the mother plants.
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