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Plant and Cell Physiology Advance Access originally published online on April 15, 2009
Plant and Cell Physiology 2009 50(5):986-997; doi:10.1093/pcp/pcp051
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© The Author 2009. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved.
The online version of this article has been published under an open access model. Users are entitled to use, reproduce, disseminate, or display the open access version of this article for non-commercial purposes provided that: the original authorship is properly and fully attributed; the Journal and the Japanese Society of Plant Physiologists are attributed as the original place of publication with the correct citation details given; if an article is subsequently reproduced or disseminated not in its entirety but only in part or as a derivative work this must be clearly indicated. For commercial re-use, please contact journals.permissions@oxfordjournals.org

Phospholipid Signaling Responses in Salt-Stressed Rice Leaves

Essam Darwish1,2, Christa Testerink1, Mohamed Khalil2, Osama El-Shihy2 and Teun Munnik1,*

1Section of Plant Physiology, Swammerdam Institute for Life Sciences, University of Amsterdam, Science Park 904, 1098 XH Amsterdam, The Netherlands
2Plant Physiology Section, Botany Department, Faculty of Agriculture, Cairo University, Egypt

*Corresponding author: E-mail, t.munnik{at}uva.nl; fax, +31(0)20-5257934.


  Abstract

Salinity is one of the major environmental factors limiting growth and productivity of rice plants. In this study, the effect of salt stress on phospholipid signaling responses in rice leaves was investigated. Leaf cuts were radiolabeled with 32P-orthophosphate and the lipids extracted and analyzed by thin-layer chromatography, autoradiography and phosphoimaging. Phospholipids were identified by co-migration of known standards. Results showed that 32Pi was rapidly incorporated into the minor lipids, phos-phatidylinositol bisphosphate (PIP2) and phosphatidic acid (PA) and, interestingly, also into the structural lipids phosphatidylethanolamine (PE) and phosphatidylglycerol (PG), which normally label relatively slowly, like phosphatidylcholine (PC) and phosphatidylinositol (PI). Only very small amounts of PIP2 were found. However, in response to salt stress (NaCl), PIP2 levels rapidly (<30 min) increased up to 4-fold, in a time- and dose-dependent manner. PA and its phosphorylated product, diacylglyc-erolpyrophosphate (DGPP), also increased upon NaCl stress, while cardiolipin (CL) levels decreased. All other phospholipid levels remained unchanged. PA signaling can be generated via the combined action of phospholipase C (PLC) and diacylglycerol kinase (DGK) or directly via phospholipase D (PLD). The latter can be measured in vivo, using a transphosphatidylation assay. Interestingly, these measurements revealed that salt stress inhibited PLD activity, indicating that the salt stress-induced PA response was not due to PLD activity. Comparison of the 32P-lipid responses in salt-tolerant and salt-sensitive cultivars revealed no significant differences. Together these results show that salt stress rapidly activates several lipid responses in rice leaves but that these responses do not explain the difference in salt tolerance between sensitive and tolerant cultivars.

Keywords: Lipid signaling - Phosphatidic acid - Phosphoinositides - Phospholipase - Rice - Salt stress

Abbreviations: CL, cardiolipin; DAG, diacylglycerol; DGK, diacylglyerol kinase; DGPP, diacylglycerolpyrophosphate; IP2, inositol bisphosphate; IP3, inositol triphosphate; IP6, inositol hexaphosphate; O/N, overnight; PA, phosphatidic acid; PBut, phosphatidylbutanol; PC, phosphatidylcholine; PE, phos-phatidylethanolamine; PG, phosphatidylglycerol; PI, phos-phatidylinositol; PI4P, phosphatidylinositol 4-phosphate; PIP2, phosphatidylinositol bisphosphate; PLC, phospholipase C; PLD, phospholipase D; PS, phosphatidylserine; TLC, thin-layer chromatography.

(Received March 2, 2009; Accepted March 31, 2009)
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