Optimizing nutrient media conditions for continuous production of shoot biomass enriched in major medicinal constituents, amarogentin and mangiferin of endangered medicinal herb, Swertia chirayita

Biosynthesis and accumulation of Amarogentin and Mangiferin from shoot culture of endangered herb Swertia chirayita helped in rescuing its natural population along with continuous production of quality rich herbal material. Although, presence of Amarogentin and Mangiferin had already been reported, but such studies did not elaborate the significant developmental stages at two varying temperature (15 ± 1 °C and 25 ± 1 °C) in shoot cultures of S. chirayita. Different developmental stages involved throughout from callus induction to complete regeneration of plant by using shoot cultures of S. chirayita, reveal different amounts of significant medicinal compounds having high pharmacological importance like bearing anti-diabetic and anti-cancerous properties. So in the present study, different developmental stages i.e. plant segment as leaf disc explants, initiation of callus formation, callus mass development, shoots primordial, manifold shoot formation and shoot elongation with complete growth were explored for accumulation of Amarogentin and Mangiferin. The Amarogentin content was 4.72 µg/mg at 15 ± 1 °C and 4.41 µg/mg at 25 ± 1 °C whereas Mangiferin content was 15.54 µg/mg at 15 ± 1 °C and 9.70 µg/mg at 25 ± 1 °C in leaf discs provided with the medium MS + 2,4D = 1 mg/L, 6BAP = 0.5 mg/L, TDZ = 0.5 mg/L, respectively. The accumulation of Amarogentin and Mangiferin started from callus cultures differentiating into shoots and reached to the detectable amount equivalent to actual leaf explants in fully grown shoots with content of 5.79 µg/mg at 15 ± 1 °C and 5.35 µg/mg at 25 ± 1 °C whereas 15.56 µg/mg at 15 ± 1 °C and 13.15 µg/mg at 25 ± 1 °C provided with the medium MS + IBA = 3 mg/L, KN = 1 mg/L, respectively. Maximum accumulation of bioactive compounds was observed in ≈3 months old in-vitro grown shoots at 15 ± 1˚ C wherein, the content of Amarogentin was ≈8.51 folds higher and Mangiferin was ≈4.09 folds higher than the ≈3 months old green house grown shoots. So, the in-vitro raised shoots of S. chirayita enriched with marker medicinal compounds would be utilized as ready to use raw material for pharmaceutical industries for herbal drug formulations and can be utilized to transfer under natural habitats for conserving its diminishing population.