Somatic embryogenesis and plant regeneration from cell suspension cultures of Gentiana kurroo Royle.

Gentiana kurroo Royle, native to North- Western Himalayas, is one of the critically endangered perennial herbs, which is being overexploited due to its multiple medicinal uses. The goal of this work was to develop a protocol for somatic embryogenesis and efficient plant regeneration through cell suspension culture. Selected ranges of plant growth regulators were experimented for somatic embryogenesis. Light micrographs of the established suspension cell cultures of somatic embryos at different developmental stages were also observed under light microscope. Somatic embryogenesis was induced from the leaf explants and the maximum induction frequency (97.2%) was obtained on Murashige and Skoog (MS) medium supplemented with 0.5 mg/l each of NAA, BAP and TDZ. Cell suspension cultures were established in MS liquid medium containing IAA (0.5 mg/l) + BAP (1.0 mg/l) with 89-93% viable cells. Transfer of cotyledonary somatic embryos to the MS agar medium containing 2, 4-D (0.4 mg/l) and KN (1.5 mg/l) resulted in somatic embryogenesis at high frequencies (80%) with an average of 28 ± 0.2 embryos/ callus piece. After transfer onto the half strength MS medium without growth regulators approximately 80-90% somatic embryos developed into complete plantlets. The protocol described here could be used for somatic hybridization, genetic transformation, isolation of protoplasts and large-scale propagation of G. kurroo.