Grape marc extract causes early perception events, defence reactions and hypersensitive response in cultured tobacco cells

• Published in: Plant physiology and biochemistry 2014-04, Vol.77, p.84-89
• Main Authors: Benouaret, R., Goujon, E., Goupil, P.
• Format: Article
• Language: English
• Subjects: Aprotinin - pharmacology; Biological and medical sciences; Cell Death; Cells, Cultured; Cycloheximide - pharmacology; Disease Resistance - genetics; Environmental Sciences; Fundamental and applied biological sciences. Psychology; Genes, Plant; Grape marc; Hydrogen-Ion Concentration; Hypersensitive reaction; Leucine - analogs & derivatives; Leucine - pharmacology; Life Sciences; Nicotiana - drug effects; Nicotiana - genetics; Nicotiana - metabolism; Pepstatins - pharmacology; Peptide Hydrolases - metabolism; Plant defence; Plant Diseases; Plant Extracts - pharmacology; Plant physiology and development; Plant Proteins - metabolism; Protease inhibitors; Tobacco BY-2; Up-Regulation; Vegetal Biology; Vitis
• ISSN: 0981-9428; 1873-2690
• DOI: 10.1016/j.plaphy.2014.01.021

Grape marc extract (GME) showed elicitor activity on suspension-cultured cells of tobacco. The BY-2 cells reacted to GME (0.25% and 0.125%) with a long-sustained pH rise in their growth medium. Using EGTA or LaCl3, we showed that extracellular alkalinization depended on Ca2+ mobilization. The tobacco BY-2 cells challenged with GME promoted cell death and the upregulation of defence-related genes such as PR3, PAL and CCoAOMT. Cell death rate was quantified using an experimental calibrated Evans Blue assay. The GME-induced cell death was dose-dependent and occurred in 24 h. Longer exposure increased the extent of tobacco cell death. To investigate a potential hypersensitive reaction, we tested the effect of various inhibitors of protein synthesis (cycloheximide) and proteases (aprotinin, pepstatin and E-64) on GME-induced cell death. All these chemicals reduced GME-induced cell death rate in 30 min. Overall, our findings indicate that GME elicits early perception events, defence reactions and cell death requiring protein synthesis and proteases. [Display omitted] •A polyphenol-rich extract from grapes induced chemoperception in tobacco suspension cells.•GME induced modification of plasma membrane properties.•GME elicited defence genes in tobacco cells.•GME-induced cell death was mediated by proteases and de novo protein synthesis.•This study evidences the ability of GME to induce hypersensitive response.