An efficient plant regeneration protocol of an industrially important plant, Hedychium coronarium J. Koenig and establishment of genetic & biochemical fidelity of the regenerants

Hedychium coronariwn is a valuable medicinal plant which is also known for its uses in cosmetics and perfumes industries. Due to high demand and unsustainable harvesting, this plant is becoming threatened in different regions of India. Tissue culture mediated plant regeneration is an alternative platform for large scale production of such plants to meet the commercial demand of the plant and plant materials in a sustainable manner. Here an efficient and improved in vitro plant regeneration protocol was established for H. coronariwn along with the genetic and biochemical fidelity analysis of the regenerants. MS medium augmented with different types, concentrations, and combinations of plant growth regulators were tested in this study. The highest number of shoot regeneration was observed from rhizome segments on Murashige and Skoog's (1962) (MS) basal medium supplemented with 0.8 mg/L thidiazuron (TDZ) (shoot induction medium) followed by their sub-culture on MS containing 1.0 mg/L gibberellic acid (GA(3)) (shoot elongation medium). Up-scaling of shoots was carried out using axenic stem segments, isolated from primary in vitro shoots, on a fresh medium of the same composition. Roots developed simultaneously during shoot multiplication and thus could eliminate the requirement of an additional step of rooting. Using this protocol ca. 540 plantlets were produced starting from single explant within 14 weeks. About 95% of these plantlets were successfully acclimatized and eventually established in the field. The micropropagated plants were morphologically similar to the mother plant. Genetic fidelity analysis of the field established micropropagated plants with that of the mother plant were carried out by using Inter Simple Sequence Repeat (ISSR) and monomorphic banding profile confirmed the genetic uniformity of the regenerants. Biochemical fidelity of the tissue culture raised plants was also validated by quantitative phytochemical analysis and High Performance Thin Layer Chromatography (HPTLC). This efficient plant regeneration protocol could be useful for commercial propagation and conservation of H. coronarium.