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A comparative proteomic analysis of tomato leaves in response to waterlogging stress
Author(s) -
Ahsan Nagib,
Lee DongGi,
Lee SangHoon,
Kang Kyu Young,
Bahk Jeong Dong,
Choi Myung Suk,
Lee InJung,
Renaut Jenny,
Lee ByungHyun
Publication year - 2007
Publication title -
physiologia plantarum
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.351
H-Index - 146
eISSN - 1399-3054
pISSN - 0031-9317
DOI - 10.1111/j.1399-3054.2007.00980.x
Subject(s) - rubisco , coomassie brilliant blue , chemistry , biochemistry , western blot , proteomics , chromatography , photosynthesis , biology , staining , gene , genetics
A comparative proteomic approach has been adopted in combination with physiological and biochemical analysis of tomato leaves responding to waterlogging stress. Waterlogging resulted in increases of relative ion leakage, lipid peroxidation and in vivo H 2 O 2 content, whereas the chlorophyll content was decreased. Histocytochemical investigations with 3,3′‐diaminobenzidine to localize H 2 O 2 and Evans blue to detect dead cells suggested that oxidative stress has a significant role to leaf senescence. Ribulose‐1,5‐bisphosphate carboxylase/oxygenase (Rubisco), the most abundant leaf protein, was successfully reduced from the samples by a fractionation method based on 15% polyethylene glycol (PEG). Elimination of Rubisco was further confirmed by Western blot analysis. To elucidate the temporal changes of the protein patterns in tomato leaves, the total soluble and the PEG‐fractionated proteins were separated by two‐dimensional electrophoresis (2‐DE) and visualized by Coomassie Brilliant Blue staining. A total of 52 protein spots were differentially expressed, wherein 33 spots were identified by matrix‐assisted laser desorption ionization time‐of‐flight (MALDI‐TOF) mass spectrometry or electrospray ionization tandem mass spectrometry (ESI‐MS/MS) analysis. The identified proteins are involved in several processes, i.e. photosynthesis, disease resistance, stress and defense mechanisms, energy and metabolism and protein biosynthesis. Results from 2‐DE analysis, combined with immunoblotting clearly showed that the fragments of Rubisco large subunit were significantly degraded. This could result from a higher production of reactive oxygen species in leaves under waterlogging stress. Furthermore, four differentially accumulated proteins were analyzed at the mRNA level, confirming the differential gene expression levels and revealing that transcription levels are not always concomitant to the translation level. A number of novel proteins were differentially expressed or appeared only in the PEG‐fractionated protein samples, indicating that PEG fractionation system can be used as a versatile protein fractionation technique in proteomic analysis to identify novel or low‐abundant proteins from all kinds of plant species.