ORIGINAL PAPER Increased cardenolides production by elicitation of Digitalis lanata shoots cultured in temporary immersion systems Naivy Pe ´rez-Alonso • Alina Capote • Andre ´ Gerth • Elio Jime ´nez Received: 30 November 2011 / Accepted: 25 February 2012 / Published online: 13 March 2012 Ó Springer Science+Business Media B.V. 2012 Abstract Digitalis lanata is an important source of car- denolides such as digoxin and lanatoside C, which have been widely applied in the treatment of cardiac insuffi- ciencies. Elicitation is one of the most effective methods to enhance the biosynthesis of several secondary metabolites in medicinal plants. We studied the effect of elicitation with Chitoplant Ò , Silioplant Ò and methyl jasmonate on biomass and cardenolides accumulation in shoots of D. lanata cultivated in temporary immersion systems. Mor- phological response of the shoots was influenced by elici- tors. A reduction in length and number of shoots was evident with all MJ concentrations. Regarding biomass production, Chitoplant Ò (0.1 g l -1 ) was found to impact significantly on fresh and dry weight of the shoots. HPLC analysis revealed a higher content of lanatoside C com- pared to digoxin in all treatments. The highest accumula- tion of lanatoside C was achieved with Chitoplant Ò (0.1 g l -1 ), which resulted in 316 lg g-DW -1 and with Silioplant Ò (0.01 g l -1 ; 310 lg g-DW -1 ), which accoun- ted for a 2.2-fold increase in lanatoside C content com- pared to non-elicited shoot cultures. Additionally, elicitation of D. lanata shoots in temporary immersion systems resulted in an oxidative stress characterized by hydrogen peroxide and malondialdehyde accumulation. These observations point to a connection between hydro- gen peroxide generation, lipid peroxidation and cardeno- lide accumulation. The optimization of elicitor treatment and culture conditions for cardenolide production as well as the advantages of TIS for this purpose are discussed. Keywords Cardiotonic glycosides Á Chitoplant Á Silioplant Á Methyl jasmonate Á Temporary immersion systems Á Oxidative stress Abbreviations 6-BAP 6-Benzylaminopurine ChP Chitoplant Ò DW Dry weight FW Fresh weight IAA Indole-3-acetic acid MDA Malondialdehyde MJ Methyl jasmonate MS Murashige and Skoog medium ROS Reactive oxygen species SiP Silioplant Ò TCA Trichloroacetic acid TBA Thiobarbituric acid TIS Temporary immersion systems Introduction In vitro culture of Digitalis species is a useful alternative for the production of therapeutically interesting cardeno- lides. Isolated cardenolides show similar pharmacody- namic properties but only a few, such as digitoxin, digoxin and lanatosides are used in humans for the treatment of cardiac insufficiency (Hornberger et al. 2000). Plants still are the sole source for their acquisition (Kuate et al. 2008). Many biotechnological strategies have been developed to enhance the production of valuable cardenolides from Digitalis. Some of these include media modification N. Pe ´rez-Alonso Á A. Capote Á E. Jime ´nez (&) Instituto de Biotecnologı ´a de las Plantas, Universidad Central ‘‘Marta Abreu’’ de Las Villas, Carretera a Camajuanı ´ km 5.5, 54830 Santa Clara, Cuba e-mail: ejimenez@ibp.co.cu A. Gerth BioPlanta GmbH, Deutscher Platz 5, 04103 Leipzig, Germany 123 Plant Cell Tiss Organ Cult (2012) 110:153–162 DOI 10.1007/s11240-012-0139-4