The present research focused on enhancing the production of wedelolactone through cell suspension culture (CSC) in Eclipta alba (L.) Hassk. With an aim of attaining a sustainable CSC, various plant growth regulators, elicitors and agitation speed were examined. Nodal segments of in vitro propagated plantlets induced the maximum percentage (93.47 ± 0.61%) of callus inoculated on Murashige and Skoog (MS) medium fortified with picloram (2 mg L−1). The growth kinetics of CSC exhibited a sigmoid pattern with a lag phase (0–6 days), a log phase (6–18 days), a stationary phase (18–24 days) and then death phase thereafter. The highest biomass accumulation in CSC with 7.09 ± 0.06 g 50 mL−1 fresh weight, 1.52 ± 0.02 g 50 mL−1 dry cell weight, 1.34 ± 0.01 × 106 cell mL−1 total cell count and 57.00 ± 0.58% packed cell volume was obtained in the liquid MS medium supplemented with 1.5 mg L−1 picloram plus 0.5 mg L−1 kinetin at 120 rpm. High performance thin layer chromatography confirmed that yeast extract (biotic elicitor) at 150 mg L−1 accumulated more CSC biomass with 1.22-fold increase in wedelolactone (288.97 ± 1.94 µg g−1 dry weight) content in comparison to the non-elicited CSC (237.78 ± 0.04 µg g−1 dry weight) after 120 h of incubation. Contrastingly, methyl jasmonate (abiotic elicitor) did not alter the biomass but increased the wedelolactone content (259.32 ± 1.06 µg g−1 dry weight) to an extent of 1.09-fold at 100 µM. Complete plantlet regeneration from CSC was possible on MS medium containing N6-benzyladenine (0.75 mg L−1) and abscisic acid (0.5 mg L−1). Thus, the establishment of protocol for CSC constitutes the bases for future biotechnological improvement studies in this crop.
Plant Cell, Tissue and Organ Culture – Springer Journals
Published: May 31, 2018
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