Tomato (Solanum lycopersicum cv. Money Maker) seedlings at the two-leaf stage were grown in one-half strength Hoagland solution supplemented with 50 mm NaCl for 4 days, with 100 mm NaCl for 4 days, with 150 mm NaCl for 4 days, and with a final concentration 200 mm NaCl for 2 days. Solutions were refreshed every 2 days for treated and untreated seedlings. Non-treated plants were grown in nonamended one-half strength Hoagland solution. Three biological replicates (BR) were included for treated and control experiments. At the end of treatments, the uppermost three newly expanded leaves from all 12 plants in each BR were collected and bulked to extract total protein. Proteomic analysis resulted in the identification of several salt-induced and salt-suppressed proteins. Salt-induced proteins were: vacuolar H+-ATPase A1 subunit isoform (1.6-fold), germin-like protein (1.5-fold), ferredoxin-NADP (+) reductase (1.2-fold), quinone oxidoreductase-like protein (4.4-fold), heat-shock protein (4.9-fold), and pyrophosphorylase (1.7-fold). Salt-suppressed proteins were: ATPase alpha subunit (−1.5-fold) and rubisco activase (−1.4-fold). Proteins identified in this study affect cellular activities for antioxidant, stress protection, carbon fixation, and carbohydrate partitioning in young tomato leaves under salt stress.
Suping Zhou, Roger Sauve, Tara Fish and Theodore W. Thannhauser
Suping Zhou, Marsha Palmer, Jing Zhou, Sarabjit Bhatti, Kevin J. Howe, Tara Fish and Theodore W. Thannhauser
A comparative proteomics study using isobaric tags for relative and absolute quantitation (iTRAQ) was performed on a mesophytic tomato (Solanum lycopersicum) cultivar and a dehydration-resistant wild species (Solanum chilense) to identify proteins that play key roles in tolerance to water deficit stress. In tomato ‘Walter’ LA3465, 130 proteins were identified, of which 104 (80%) were repressed and 26 (20%) were induced. In S. chilense LA1958, a total of 170 proteins were identified with 106 (62%) repressed and 64 (38%) induced. According to their putative molecular functions, the differentially expressed proteins belong to the following subgroups: stress proteins, gene expression, nascent protein processing, protein folding, protein degradation, carbohydrate metabolism, amino acid and nucleotide metabolism, lipid metabolism, signal transduction, and cell cycle regulation. Based on changes in protein abundance induced by the dehydration treatment, cellular metabolic activities and protein biosynthesis were suppressed by the stress. In S. chilense, dehydration treatment led to elevated accumulation of proteins involved in post-transcriptional gene regulation and fidelity in protein translation including prefoldin, which promotes protein folding without the use of adenosine-5′-triphosphate (ATP), several hydrophilic proteins, and calmodulin in the calcium signal transduction pathway. Those protein changes were not found in the susceptible tomato, ‘Walter’. Within each functional protein group, proteins showing opposite changes (dehydration induced vs. repressed) in the two species were identified and roles of those proteins in conferring tolerance to water deficit stress are discussed. Information provided in this report will be useful for selection of proteins or genes in analyzing or improving dehydration tolerance in tomato cultivars.
Suping Zhou, Roger J. Sauvé, Zong Liu, Sasikiran Reddy, Sarabjit Bhatti, Simon D. Hucko, Tara Fish and Theodore W. Thannhauser
This article reports salt-induced changes in leaf and root proteomes after wild tomato (Solanum chilense) plants were treated with 200 mm NaCl. In leaf tissues, a total of 176 protein spots showed significant changes (P < 0.05), of which 104 spots were induced and 72 spots suppressed. Salt-induced proteins are associated with the following pathways: photosynthesis, carbohydrate metabolism, glyoxylate shunt, glycine cleavage system, branched-chain amino acid biosynthesis, protein folding, defense and cellular protection, signal transduction, ion transport, and antioxidant activities. Suppressed proteins belong to the following categories: oxidative phosphorylation pathway, photorespiration and protein translational machinery, oxidative stress, and ATPases. In root tissues, 106 protein spots changed significantly (P < 0.05) after the salt treatment, 63 spots were induced, and 43 suppressed by salt treatment. Salt-induced proteins are associated with the following functional pathways: regeneration of S-adenosyl methionine, protein folding, selective ion transport, antioxidants and defense mechanism, signal transduction and gene expression regulation, and branched-chain amino acid synthesis. Salt-suppressed proteins are receptor kinase proteins, peroxidases and germin-like proteins, malate dehydrogenase, and glycine dehydrogenase. In this study, different members of proteins were identified from leaf and root tissues after plants were subjected to salt treatment. These proteins represent tissue-specific changes in salt-induced proteomes. When protein expression was compared in the context of metabolic pathways, the branched-chain amino acid biosynthesis, glucose catabolism toward reducing cellular glucose level, and the antioxidant, detoxification, and selective ion uptake and transport were induced in both root and leaf tissues. These changes appear to be associated with salt tolerance in the whole plant.
Peter Nveawiah-Yoho, Jing Zhou, Marsha Palmer, Roger Sauve, Suping Zhou, Kevin J. Howe, Tara Fish and Theodore W. Thannhauser
Tomato (Solanum lycopersicum) has a wide variety of genotypes differing in their responses to salinity. This study was performed to identify salt-induced changes in proteomes that are distinguishable among tomatoes with contrasting salt tolerance. Tomato accessions [LA4133 (a salt-tolerant cherry tomato accession) and ‘Walter’ LA3465 (a salt-susceptible accession)] were subjected to salt treatment (200 mm NaCl) in hydroponic culture. Salt-induced changes in the root proteomes of each tomato accession were identified using the isobaric tags for relative and absolute quantitation (iTRAQ) method. In LA4133, 178 proteins showed significant differences between salt-treated and non-treated control root tissues (P ≤ 0.05); 169 proteins were induced (1.3- to 5.1-fold) and nine repressed (–1.7- to –1.3-fold). In LA3465, 115 proteins were induced (1.3- to 6.4-fold) and 23 repressed (–2.5- to –1.3-fold). Salt-responsive proteins from the two tomato accessions were involved in the following biological processes: root system development and structural integrity; carbohydrate metabolism; adenosine-5′-triphosphate regeneration and consumption; amino acid metabolism; fatty acid metabolism; signal transduction; cellular detoxification; protein turnover and intracellular trafficking; and molecular activities for regulating gene transcription, protein translation, and post-translational modification. Proteins affecting diverse cellular activities were identified, which include chaperonins and cochaperonins, heat-shock proteins, antioxidant enzymes, and stress proteins. Proteins exhibiting different salt-induced changes between the tolerant and susceptible tomato accessions were identified, and these proteins were divided into two groups: 1) proteins with quantitative differences because they were induced or repressed by salt stress in both accessions but at different fold levels; and 2) proteins showing qualitative differences, where proteins were induced in one vs. repressed or not changed in the other accession. Candidate proteins for tolerance to salt and secondary cellular stresses (such as hypo-osmotic stress and dehydration) were proposed based on findings from the current and previous studies on tomato and by the use of the Arabidopsis thaliana protein database. Information provided in this report will be very useful for evaluating and breeding for plant tolerance to salt and/or water deficit stresses.
Suping Zhou, Roger J. Sauvé, Zong Liu, Sasikiran Reddy, Sarabjit Bhatti, Simon D. Hucko, Yang Yong, Tara Fish and Theodore W. Thannhauser
Three tomato (Solanum lycopersicum) cultivars [Walter LA3465 (heat-tolerant), Edkawi LA 2711 (unknown heat tolerance, salt-tolerant), and LA1310 (cherry tomato)] were compared for changes in leaf proteomes after heat treatment. Seedlings with four fully expanded leaves were subjected to heat treatment of 39/25 °C at a 16:8 h light–dark cycle for 7 days. Leaves were collected at 1200 hr, 4 h after the light cycle started. For ‘Walter’ LA3465, heat-suppressed proteins were geranylgeranyl reductase, ferredoxin-NADP (+) reductase, Rubisco activase, transketolase, phosphoglycerate kinase precursor, fructose–bisphosphate aldolase, glyoxisomal malate dehydrogenase, catalase, S-adenosyl-L-homocysteine hydrolase, and methionine synthase. Two enzymes were induced, cytosolic NADP-malic enzyme and superoxide dismutase. For ‘Edkawi’ LA2711, nine enzymes were suppressed: ferredoxin-NADP (+) reductase, Rubisco activase, S-adenosylmethionine synthetase, methioine synthase, glyoxisomal malate dehydrogenase, enolase, flavonol synthase, M1 family peptidase, and dihydrolipoamide dehydrogenase. Heat-induced proteins were cyclophilin, fructose-1,6-bisphosphate aldolase, transketolase, phosphoglycolate phosphatase, ATPase, photosystem II oxygen-evolving complex 23, and NAD-dependent epimerase/dehydratase. For cherry tomato LA1310, heat-suppressed proteins were aminotransferase, S-adenosyl-L-homocysteine hydrolase, L-ascorbate peroxidase, lactoylglutathione lyase, and Rubisco activase. Heat-induced enzymes were glyoxisomal malate dehydrogenase, phosphoribulokinasee, and ATP synthase. This research resulted in the identification of proteins that were induced/repressed in all tomato cultivars evaluated (e.g., Rubisco activase, methionine synthase, adenosyl-L-homocysteine hydrolase, and others) and those differentially expressed (e.g., transketolase).