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Journal of Biological Sciences

Year: 2003 | Volume: 3 | Issue: 11 | Page No.: 994-1009
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Research Article

Involvement of Protein Phosphorylation and Reactive Oxygen Species in Jasmonate-elicited Accumulation of Defense/stress-related Proteins in Rice Seedlings

Randeep Rakwal, Md. Monowar Karim Khan and Setsuko Komatsu

ABSTRACT

Calcium chelators/antagonist effectively blocked the jasmonic acid (JA) induced accumulation of a basic 28 kDa Bowman-Birk proteinase inhibitor and an acidic 17 kDa pathogenesis-related class 1 protein in rice seedling leaf sheaths. In gel kinase assays revealed rapid changes in the phosphorylation activities of a 46 and 56 kDa Myelin Basic Protein Kinase (MBPK) and a 48 kDa putative Ca2+-dependent protein kinase (CDPK) by JA and leaf sheath cutting and these changes were determined to be cytosolic in nature. In vitro protein phosphorylation of crude and cytosolic protein extracts, followed by two-dimensional polyacrylamide gel electrophoresis showed considerable enhancement in the Ca2+-dependent phosphorylation of certain proteins as early as 15 min after JA treatment, particularly a 56 kDa phosphoprotein. Hydrogen peroxide (a potent reactive oxygen species, ROS) and oxyfluorfen (a lipid peroxidizing agent) caused a significant increase in the levels of these two proteins over the cut control. Whereas, when JA was co-applied with 3-(3`, 4`-dichlorophenyl)-1,1-dimethylurea, a photosystem II electron transport inhibitor and n-propyl gallate, a free radical scavenger, the accumulation of these induced proteins were considerably reduced. These results suggest an involvement of kinase-signaling cascade(s) and the ROS in JA elicited accumulation of these defense/stress-related proteins in rice.
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