A proteomic screen and identification of waterlogging-regulated proteins in tomato roots

Abstract

To gain a better understanding of the tomato root response to waterlogging stress at protein level, a proteomic approach has been conducted in combination with physiological analyses. Fiveweek old plants were subjected to waterlogging stress, and roots were collected 24 and 72 h after treatment. Waterlogging stress resulted in a gradual increase of lipid peroxidation and in vivo H 2 O 2 concentration in roots. To elucidate temporal changes in protein expression in tomato roots, proteins were extracted from the root samples, separated by 2-DE, and visualized by staining with Coomassie Brilliant Blue. A total of 35 proteins were diVerentially expressed in tomato roots in response to waterlogging stress, and 29 were identiWed using MALDI-TOF analysis. Of the proteins identiWed, 16 proteins were up-regulated and 13 proteins were down-regulated in waterlogged tomato roots. In addition to previously identiWed waterlogging-regulated proteins, we identiWed several novel proteins, including 3-beta-hydroxylase, phenylalanine ammonia-lyase, glutamyl-tRNA reductase, Xavanone 3-hydroxylase, mitochondrial ATPase alpha subunit, cysteine protease, DWARF1, and NIM1-like protein 2. These newly identiWed proteins are involved in several cellular processes, including hormone and secondary metabolite synthesis, programmed cell death, and stress and defense mechanisms. Five diVerentially accumulated proteins were further analyzed at the mRNA level, which conWrmed the diVerential gene expression, and revealed that the transcript levels were not always consistent with the translation level. The identiWcation of these novel proteins in the waterlogging stress response provides new insight that can lead to a better understanding of the molecular basis of waterlogging-sensitivity in plants. Abbreviations 2-DETwo-dimensional gel electrophoresis ANP Anaerobic polypeptide BR Brassinosteroid MALDI-TOF Matrix assisted laser desorption ionization time-of-Xight PAL Phenylalanine ammonia-lyase PEG Polyethylene glycol ROS Reactive oxygen species SAR Systemic acquired resistance SDS-PAGE Sodium dodecylsulfate polyacrylamide gel electrophoresis TBARS Thiobarbituric acid reactive substance

Proteomic response of wheat embryos to fosthiazate stress in a protected vegetable soil

“…Metabolism-related proteins were found to be significantly up-or down-regulated during germination in wheat, as detected in tomato, rice and cucumber (Ahsan et al, 2007b(Ahsan et al, , 2007cDu et al, 2010). In the present study we detected differential expression of proteins involved in carbohydrate, fatty acid and amino acid metabolism.…”

“…2-DE is used to investigate differentially-expressed proteins in response to abiotic or biotic stresses such as heavy metals (Ahsan et al, 2007a(Ahsan et al, , 2007c, herbicides (Ahsan et al, 2008;Castro et al, 2005;Lin et al, 2008), flooding (Ahsan et al, 2007b;Kong et al, 2010), salt stress (Du et al, 2010), chilling , ovine saliva , Meloidogyne incognita (de Deus Barbosa, 2009) and pathogens (Lee, 2006). It is well-documented that the germination process is highly disturbed by heavy metals (Xu et al, 2009), herbicides (Song et al, 2007) and antibiotics (Jin et al, 2010).…”

Proteomic identification of differentially expressed proteins in the anoxic rice coleoptile

“…Proteomic studies on plants exposed to low oxygen stress have shown that, in addition to the expression of classical anaerobic proteins (ANPs), there is also expression of proteins related to other mechanisms, such as reactive oxygen species (ROS) scavengers and protein synthesis in wheat (Kong et al, 2010), heat stress response in soybean (Hashiguchi et al, 2009), intracellular trafficking in maize (Chang et al, 2000), gibberellin biosynthesis, heme biosynthesis, and cell wall degradation in tomatoes (Ahsan et al, 2007).…”

Proteome analysis of soybean roots under waterlogging stress at an early vegetative stage

“…Dramatic changes in protein synthesis have been reported in plant roots during anaerobiosis. Most of these anaerobic proteins (ANPs) have been identifi ed as enzymes of glycolysis or sugar phosphate metabolism (Sachs et al 1996;Ahsan et al 2007a). ANPs that are part of other metabolic processes have also been reported (Chang et al 2000;Shi et al 2008).…”