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Feeding elicitors and precursors enhance colchicine accumulation in morphogenic cultures of Gloriosa superba L.

Jawahar, G., Punita, D. L., Rajasheker, G., Manoharachary, C., Venkatachalam, P., Kavi Kishor, P. B.
Plant cell, tissue, and organ culture 2018 v.135 no.2 pp. 235-245
2,4-D, Alternaria solani, Bacillus subtilis, Fusarium oxysporum, Gloriosa superba, Saccharomyces cerevisiae, activated carbon, aluminum chloride, aspirin, biosynthesis, colchicine, elicitors, leaves, mercuric chloride, naphthaleneacetic acid, nitroprusside, phenylalanine, rubidium, salicylic acid, sodium chloride, stress response, sucrose, tryptophan, tubers, tyrosine
Morphogenic cultures of Gloriosa superba were initiated on Murashige and Skoog’s medium fortified with 2 mg L⁻¹ 2,4-dichlorophenoxyacetic acid (2,4-D), 0.5 mg L⁻¹ naphthaleneacetic acid (NAA), 4% sucrose and 0.1% activated charcoal. To enhance the content of the alkaloid colchicine, morphogenic cultures were treated with different concentrations of abiotic elicitors like signalling compounds, metals, biotic elicitors, precursors and a combination of elicitors. Signalling molecules like acetyl salicylic acid (ASA) and sodium nitroprusside improved the production of colchicine. Abiotic elicitors have markedly (p ≤ 0.05 or ≤ 0.01) enhanced the colchicine content either at lower or higher concentrations. Among the metals, the highest amount of 11.67 mg of colchicine g⁻¹ dry wt was noticed at 60 mM rubidium chloride, followed by 60 mM NaCl (11.18 mg g⁻¹). Contrarily, in the presence of biotic elicitors such as Fusarium oxysporum, Alternaria solani, and Saccharomyces cerevisiae, colchicine content ranged only between 2 and 5.32 mg g⁻¹, but Bacillus subtilis repressed it. Among the aromatic amino acids, phenylalanine at 500 mg L⁻¹ influenced the highest accumulation of 19.48 mg g⁻¹ dry tissue, followed by tryptophan (12.47 mg g⁻¹), and tyrosine (9.87 mg g⁻¹), a direct precursor of colchicine biosynthesis, while intact tubers and leaves contained 4.65 and 4.16 mg of colchicine g⁻¹ dry tissue respectively. A combination of 10 µM AlCl₃ and 50 µM salicylic acid (SA) registered 17.34 mg g⁻¹ followed by 16.24 mg g⁻¹ tissue in presence of 1 µM HgCl₂ and 50 µM SA. The results suggest that the elicitor-stimulated colchicine accumulation was a stress response and can be exploited further for commercial production.