Quantitative proteomic analysis of Cabernet Sauvignon grape cells exposed to thermal stresses reveal alterations in sugar and phenylpropanoid metabolism — ASN Events
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Quantitative proteomic analysis of Cabernet Sauvignon grape cells exposed to thermal stresses reveal alterations in sugar and phenylpropanoid metabolism (#105)

Iniga S George 1 , Dana Pascovici 2 , Mehdi Mirzaei 1 , Paul A Haynes 1
  1. Macquarie University, Sydney, NSW, Australia
  2. Australian Proteome Analysis Facility (APAF), Macquarie University , Sydney, NSW, Australia

Grapes (Vitis vinifera) are a valuable fruit crop and wine production is a major industry. Global warming and expanded range of cultivation will expose grapes to more temperature stresses in future. We investigated quantitative shotgun proteomic changes induced by temperature stresses on Cabernet Sauvignon cells to provide insights into metabolic pathways related to temperature stress in grapevines. Our study examined proteomic changes induced by the impact of four different temperature stress regimes, including both hot and cold temperatures, on cultured grape cells. Cabernet Sauvignon grape cell suspension cultures grown at 26°C were subjected to 14 hours of exposure to 34°C and 42°C for heat stress, and 18°C and 10°C for cold stress. Cells from the five temperatures were harvested in biological triplicates. Proteins were extracted using methanol-chloroform [1] and digested by filter aided sample preparation (FASP) protocol [2]. Proteins were identified with a Velos-Pro linear ion-trap mass spectrometer by gas phase fractionation (GPF) [3], and spectra were searched against UniProtKB Vitis vinifera database and quantified by spectral counting. A total of 1755 non-redundant proteins were identified from the five temperature points. Ninety-eight proteins were identified exclusively in extreme heat stress (42°C) and ninety-five proteins were identified exclusively at extreme cold stress (10°C). Gene Ontology (GO) annotations of differentially expressed proteins provided insights into the metabolic pathways that are involved in temperature stress responses in grape cells. Sugar metabolism displayed switching between alternative and classical pathways during extreme temperature stresses. Nine proteins involved in the phenylpropanoid pathway were highly up-regulated at extreme cold stress, and numerous other proteins involved in phenylpropanoid metabolism and anthocyanin synthesis were found to be cold-responsive proteins. This is the first label-free shotgun proteomic study on grapes exposed to hot and cold temperature stress.

  1. Wessel, D., Flugge, U. I., A Method for the Quantitative Recovery of Protein in Dilute Solution in the Presence of Detergents and Lipids. Analytical Biochemistry 1984, 138, 141-143.
  2. Chapman, B., Castellana, N., Apffel, A., Ghan, R., et al., Plant proteogenomics: from protein extraction to improved gene predictions. Methods in molecular biology 2013, 1002, 267-294.
  3. Scherl, A., Shaffer, S. A., Taylor, G. K., Kulasekara, H. D., et al., Genome-specific gas-phase fractionation strategy for improved shotgun proteomic profiling of proteotypic peptides. Analytical chemistry 2008, 80, 1182-1191.