A hydroponic rice seedling culture model system for investigating proteome of salt stress in rice leaf.
By using an in vivo hydroponic rice seedling culture system, we investigated the physiological and biochemical responses of a model rice japonica cultivar Nipponbare to salt stress using proteomics and classical biochemical methods. Yoshida's nutrient solution (YS) was used to grow rice seedlings. YS-grown 18-day-old seedlings manifested highly stable and reproducible symptoms, prominently the wilting and browning of the 3rd leaf, reduced photosynthetic activity, inhibition in overall seedling growth, and failure to develop new (5th) leaf, when subjected to salt stress by transferring them to YS containing 130 mM NaCl for 4 days. As leaf response to salt stress is least investigated in rice by proteomics, we used the 3rd leaf as source material. A comparison of 2-DE protein profiles between the untreated control and salt-stressed 3rd leaves revealed 55 differentially expressed CBB-stained spots, where 47 spots were increased over the control. Of these changed spots, the identity of 33 protein spots (27 increased and 5 decreased) was determined by nESI-LC-MS/MS. Most of these identified proteins belonged to major metabolic processes like photosynthetic carbon dioxide assimilation and photorespiration, suggesting a good correlation between salt stress-responsive proteins and leaf morphology. Moreover, 2-DE immunoblot and enzymatic activity analyses of 3rd leaves revealed remarkable changes in the key marker enzymes associated with oxidative damage to salt stress: ascorbate peroxidase and lipid peroxidation were induced, and catalase was suppressed. These results demonstrate that hydroponic culture system is best suited for proteomics of salt stress in rice seedling.
Kim
Dea-Wook
DW
Graduate School of Life and Environmental Science, University of Tsukuba, Tsukuba, Ibaraki, Japan. deawook@hotmail.com
Rakwal
Randeep
R
Agrawal
Ganesh Kumar
GK
Jung
Young-Ho
YH
Shibato
Junko
J
Jwa
Nam-Soo
NS
Iwahashi
Yumiko
Y
Iwahashi
Hitoshi
H
Kim
Du Hyun
DH
Shim
Ie-Sung
IeS
Usui
Kenji
K
D000595:Amino Acid Sequence; D060387:Ascorbate Peroxidases; D002374:Catalase / Q000378:metabolism; D002853:Chromatography, Liquid; D046508:Culture Techniques; D015180:Electrophoresis, Gel, Two-Dimensional; D018527:Hydroponics; D015227:Lipid Peroxidation; D008969:Molecular Sequence Data; D012275:Oryza / Q000254:growth & development / Q000378:metabolism*; D009997:Osmotic Pressure; D010084:Oxidation-Reduction; D010544:Peroxidases / Q000378:metabolism; D018515:Plant Leaves / Q000378:metabolism*; D010940:Plant Proteins / Q000378:metabolism*; D020543:Proteome / Q000378:metabolism*; D036226:Seedlings / Q000254:growth & development / Q000378:metabolism; D012965:Sodium Chloride / Q000494:pharmacology; D021241:Spectrometry, Mass, Electrospray Ionization
D010940:Plant Proteins; D020543:Proteome; D012965:Sodium Chloride; D010544:Peroxidases; D060387:Ascorbate Peroxidases; D002374:Catalase