M. Bieb, a perennial herbaceous wild species, is commonly used in the horticultural, ornamental, and pharmaceutical markets. Studies on the tissue culture systems for this species would be beneficial for mass multiplication as well as for future breeding programs. An in vitro propagation technique was established here using tuberous root explants as unique and responsive starting materials for culture initiation. The proliferated calli were sub-cultured on shoot proliferation media and regenerated microshoots were assessed. The shoot proliferation rate, leaf number, leaf length, and chlorophyll and carotenoid contents were recorded. The highest callus induction per explant (76.67%), callus dry weight (10.25 mg), callus firmness ratio (3.97), and callus color intensity ratio (2.83) were observed in explants inoculated on Murashige and Skoog (MS) medium supplemented with 10.0 mgL−1
6-benzylaminopurine (BAP). The highest shoot proliferation rates were obtained when calli were sub-cultured on MS or Schenk and Hildebrandt (SH) basal media supplemented with 2.0 mgL−1
BAP. The half-strength MS medium fortified with 4.0% sucrose + 2.0 mgL−1
indole butyric acid (IBA) + 200 mgL−1
activated charcoal was a superior combination for root emergence and rooting parameters. Regenerated plantlets were then successfully adapted to ex vitro conditions. The reported protocol can be exploited at a commercial scale following minor modification, or could be beneficial in the production of secondary metabolites in bioreactors where callus is required as fresh plant material.
This is an open access article distributed under the Creative Commons Attribution License
which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited