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Phospholipase D in Phytophthora infestans and Its Role in Zoospore Encystment

September 2002 , Volume 15 , Number  9
Pages  939 - 946

Maita Latijnhouwers , 1 Teun Munnik , 2 and Francine Govers 1

1Laboratory of Phytopathology, Wageningen University, Binnenhaven 5, NL-6709 PD Wageningen and Graduate School Experimental Plant Sciences, The Netherlands; 2Swammerdam Institute for Life Sciences, University of Amsterdam, Kruislaan 318, NL-1098 SM, Amsterdam, The Netherlands

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Accepted 6 May 2002.

We show that differentiation of zoospores of the late blight pathogen Phytophthora infestans into cysts, a process called encystment, was triggered by both phosphatidic acid (PA) and the G-protein activator mastoparan. Mastoparan induced the accumulation of PA, indicating that encystment by mastoparan most likely acts through PA. Likewise, mechanical agitation of zoospores, which often is used to induce synchronized encystment, resulted in increased levels of PA. The levels of diacylglycerolpyrophosphate (DGPP), the phosphorylation product of PA, increased simultaneously. Also in cysts, sporangiospores, and mycelium, mastoparan induced increases in the levels of PA and DGPP. Using an in vivo assay for phospholipase D (PLD) activity, it was shown that the mastoparan-induced increase in PA was due to a stimulation of the activity of this enzyme. Phospholipase C in combination with diacylglycerol (DAG) kinase activity also can generate PA, but activation of these enzymes by mastoparan was not detected under conditions selected to highlight 32P-PA production via DAG kinase. Primary and secondary butanol, which, like mastoparan, have been reported to activate G-proteins, also stimulated PLD activity, whereas the inactive tertiary isomer did not. Similarly, encystment was induced by n- and sec-butanol but not by tert-butanol. Together, these results show that Phytophthora infestans contains a mastoparan- and bu-tanol-inducible PLD pathway and strongly indicate that PLD is involved in zoospore encystment. The role of G-proteins in this process is discussed.

Additional keywords: alcohols , phospholipids .

© 2002 The American Phytopathological Society