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Ergosterol-Induced Sesquiterpenoid Synthesis in Tobacco Cells
Department of Biochemistry, University of Johannesburg, Auckland Park, Johannesburg, 2006, South Africa
Drug Discovery and Development, CSIR Biosciences, Pretoria, 0001, South Africa
* Author to whom correspondence should be addressed.
Received: 23 November 2011; in revised form: 26 January 2012 / Accepted: 6 February 2012 / Published: 9 February 2012
(This article belongs to the Special Issue Terpenoids
Abstract: Plants have the ability to continuously respond to microbial signals in their environment. One of these stimuli is a steroid from fungal membranes, ergosterol, which does not occur in plants, but acts as a pathogen-associated molecular pattern molecule to trigger defence mechanisms. Here we investigated the effect of ergosterol on the secondary metabolites in tobacco (Nicotiana tabacum) cells by profiling the induced sesquiterpenoids. Suspensions of tobacco cells were treated with different concentrations (0–1,000 nM) of ergosterol and incubated for different time periods (0–24 h). Metabolites were extracted with a selective dispersive liquid-liquid micro-extraction method. Thin layer chromatography was used as a screening method for identification of sesquiterpenoids in tobacco extracts. Liquid chromatography coupled to mass spectrometry was used for quantitative and qualitative analyses. The results showed that ergosterol triggered differential changes in the metabolome of tobacco cells, leading to variation in the biosynthesis of secondary metabolites. Metabolomic analysis through principal component analysis-scores plots revealed clusters of sample replicates for ergosterol treatments of 0, 50, 150, 300 and 1,000 nM and time-dependent variation at 0, 6, 12, 18 and 24 h. Five bicyclic sesquiterpenoid phytoalexins, capsidiol, lubimin, rishitin, solavetivone and phytuberin, were identified as being ergosterol-induced, contributing to the altered metabolome.
Keywords: ergosterol; metabolomics; Nicotiana tabacum; phytoalexins; secondary metabolites; sesquiterpenoids; UPLC-HDMS
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Tugizimana, F.; Steenkamp, P.A.; Piater, L.A.; Dubery, I.A. Ergosterol-Induced Sesquiterpenoid Synthesis in Tobacco Cells. Molecules 2012, 17, 1698-1715.
Tugizimana F, Steenkamp PA, Piater LA, Dubery IA. Ergosterol-Induced Sesquiterpenoid Synthesis in Tobacco Cells. Molecules. 2012; 17(2):1698-1715.
Tugizimana, Fidele; Steenkamp, Paul A.; Piater, Lizelle A.; Dubery, Ian A. 2012. "Ergosterol-Induced Sesquiterpenoid Synthesis in Tobacco Cells." Molecules 17, no. 2: 1698-1715.