In vitro propagation of Bergenia stracheyi: an alternative approach for higher production of valuable bioactive compounds

Bergenia stracheyi, commonly known as ‘Pashanbheda’ or Zakhm-e-Hayat, is a perennial herb that has been recognized for its diverse medicinal properties. The over-exploitation of B. stracheyi has threatened this species. This research aimed to develop a robust tissue culture protocol that can be utilized for rapid micropropagation of B. stracheyi. This protocol is crucial for ensuring the sustainable production of this valuable plant species and preventing the depletion of its natural populations. This study successfully demonstrated an efficient in vitro regeneration protocol by using leaf explants. For shoot induction and multiplication, MS media were supplemented with BAP (6- benzyl amino purine), IAA (indole-3-acetic acid) and NAA (1-naphthalene acetic acid). The most positive response for callus induction was observed on MS media supplemented with BAP (1.5 mg/L) + NAA (1 mg/L). The greatest number of shoots was observed on solid MS media supplemented with BAP (2 mg/L) + IAA (2 mg/L). The maximum shoot response was obtained on MS media supplemented with BAP (2.5 mg/L). The maximum number of roots was achieved on solid MS media without growth regulators. Rooted plantlets were successfully acclimatized in pots and then transferred to an open field. An analytical method using high-performance liquid chromatography and thin layer chromatography was developed for the identification and quantification of two marker compounds (bergenin and gallic acid) in tissue culture extracts as well as in wild samples. The highest contents of bergenin (4.64 mg/mL) and gallic acid (4.29 mg/mL) were found in the methonolic shoot extracts supplemented with BAP (2.0 mg/mL), which were comparable to the amounts present in the rhizomes of wild B. stracheyi (5.54 mg/mL and 3.62 mg/mL, respectively). The integral combination of tissue culture and analytical techniques could serve as a baseline for understanding the important biosynthetic pathways of completely novel, complex and bioactive metabolites of B. stracheyi.