Callus induction and high frequency organogenesis in saffron (Crocus sativus L.)

Saffron (Crocus sativus L.) is an economically important plant with a very slow natural multiplication rate and plant tissue culture is expected to enhance its reproductive capacity. In present study, corm explants were used for micro-propagation. Auxins and cytokinin were found important for in vitro propagation of explants. Overall, 93% calli were induced from meristematic region of corm explants propagated on Murashige and Skoog (MS) medium supplemented with 2 mg L⁻¹ 2,4-dichlorophenoxy-acetic acid (2,4-D), 0.5 mg L⁻¹ kinetin (Kn), 2% sucrose and 100 mg L⁻¹ ascorbic acid after 6 weeks inoculation. Somatic embryogenesis was observed on half strength MS medium supplemented with 2 mg L⁻¹ indole-3-acetic acid, 2 mg L⁻¹ Kn and 100 mg L⁻¹ ascorbic acid. Organogenesis from calli was observed with 89.6% shooting on MS medium containing 2 mg L⁻¹ Kn and 2 mg L⁻¹ IAA. Adventitious rooting of 91.6% calli was obtained on MS medium supplemented with 2 mg L⁻¹ indole-3-butyric acid. There are plenty of protocols available for tissue culture of saffron but this protocol provides an easy and time efficient method for micropropagation of saffron in vitro.

Abbreviations: 2,4-D: 2,4-dichlorophenoxyacetic acid; BAP: 6-benzyl aminopurine, NAA: 1-naphthalene acetic acid; HgCb: mercuric chloride; NaOCl: sodium hypochlorite, KMnO 4: potassium permangnate; PPM: plant preservation mixture; TDZ: thidiazuron; MS: Murashige and Skoog medium; Kn: kinetin; IAA: indole-3-acetic acid; IBA: indole-3-butyric acid.