First published online January 22, 2004; 10.1104/pp.103.029454
Plant Physiology 134:813-823 (2004)
© 2004 American Society of Plant Biologists
CELL BIOLOGY AND SIGNAL TRANSDUCTION
Osmotically Induced Cell Swelling versus Cell Shrinking Elicits Specific Changes in Phospholipid Signals in Tobacco Pollen Tubes1
Laura Zonia* and
Teun Munnik
Institute of Experimental Botany, Na Pernikarce 15, 160 00 Prague 6, Czech Republic (L.Z.); and Swammerdam Institute for Life Sciences, Department of Plant Physiology, University of Amsterdam, Kruislaan 318, NL1098 SM Amsterdam, The Netherlands (T.M.)
Pollen tube cell volume changes rapidly in response to perturbation of the extracellular osmotic potential. This report shows that specific phospholipid signals are differentially stimulated or attenuated during osmotic perturbations. Hypo-osmotic stress induces rapid increases in phosphatidic acid (PA). This response occurs starting at the addition of 25% (v/v) water to the pollen tube cultures and peaks at 100% (v/v) water. Increased levels of PA were detected within 30 s and reached maximum by 15 to 30 min after treatment. The pollen tube apical region undergoes a 46% increase in cell volume after addition of 100% water (v/v), and there is an average 7-fold increase in PA. This PA increase appears to be generated by phospholipase D because concurrent transphosphatidylation of n-butanol results in an average 8-fold increase in phosphatidylbutanol. Hypo-osmotic stress also induces an average 2-fold decrease in phosphatidylinositol phosphate; however, there are no detectable changes in the levels of phosphatidylinositol bisphosphates. In contrast, salt-induced hyperosmotic stress from 50 to 400 mM NaCl inhibits phospholipase D activity, reduces the levels of PA, and induces increases in the levels of phosphatidylinositol bisphosphate isomers. The pollen tube apical region undergoes a 41% decrease in cell volume at 400 mM NaCl, and there is an average 2-fold increase in phosphatidylinositol 3,5-bisphosphate and 1.4-fold increase in phosphatidylinositol 4,5-bisphosphate. The phosphatidylinositol 3,5-bisphosphate increase is detected within 30 s and reaches maximum by 15 to 30 min after treatment. In summary, these results demonstrate that hypo-osmotic versus hyperosmotic perturbation and the resultant cell swelling or shrinking differentially activate specific phospholipid signaling pathways in tobacco (Nicotiana tabacum) pollen tubes.
Article, publication date, and citation information can be found at http://www.plantphysiol.org/cgi/doi/10.1104/pp.103.029454.
1 This work was supported by The Netherlands Organization for Scientific Research (grant nos. NWO:99002, 810.66.011, 81036.005, and 813.06.003 to T.M.), by The Royal Netherlands Academy of Arts and Sciences (to T.M.), by the European Commission (grant nos. HPRNCT200000093 and HPRNCT200200251 to T.M.), and by the Czech Republic (Research Center grant to L.Z., "Signaling in Plants" Ministerstvo kolství, mláde e a t lov chovy LN00A081).
* Corresponding author; e-mail zonia{at}ueb.cas.cz; fax 420233339412.
Received July 9, 2003;
returned for revision August 21, 2003;
accepted November 15, 2003.
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