Mehpara MAQSOOD, Mir KHUSRAU, Zahoor A. KALOO, Tariq A. WANI, Abdul MUJIB

Colchicine Quantification in Salt Stress Treated Culture of Colchicum luteum Baker by High Pressure Liquid Chromatography

Objective: The aim of the present work was to develop a protocol for season independent propagation of Colchicum luteum and enrichment of colchicine yield under salt stress.Materials and Methods: C. luteum was collected from Kashmir, India and was cultured in vitro. The callus was exposed to different NaCl (salt) treatments and the yield of colchicine was quantified by high pressure liquid chromatography (HPLC). Results: Different explants viz. seeds, leaves, anthers and corm were used and the callus was only induced from cormsegments. The callus induction and proliferation were best achieved on Murashige and Skoog medium supplemented with 2.0 mg L-1 2, 4-D + 4.0 mg L-1 BAP. Direct and indirect plant regeneration from corm was observed in 2.0 mg L-1 BAP + 2.0 mg L-1 NAA added medium. The addition of adjuvants like activated charcoal and citric acid was noted to be less efficient forimproving callus growth. Induced callus was elicitated with different NaCl concentrations (T0 = without NaCl, T1 = 25, T2 = 50, T3 = 75, and T4 =100 mM). The yield of colchicine was quantified by HPLC at periodic intervals (2, 4, and 6 weeks). All the used levels improved colchicine yield, but the content was maximum at T3 (871.4 ng mg-1 DWB). This is the first report of callus induction and plant regeneration from corm and the quantification of colchicine under salt stress in C. luteum. Conclusion: Sodium chloride is thus a potential inducer and elicitor in improving colchicine yield in C. luteum.

Keywords:

Colchicum luteum exudates, regeneration, colchicine, in vitro cultures, stress,

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