Role of Biosurfactant and Ion Channel-Forming Activities of Syringomycin Tranransmembrane Ion Flux: A Model for the Mechanism of Action in the Plant-Pathogen Interaction. Michael L. Hutchison . Department of Plant Pathology, Washington State University, Pullman, Washington 99164-6430 U.S.A. Mark A.Tester(2), and Dennis C. Gross(1). (1) Department of Plant Pathology, Washington State University, Pullman, Washington 99164-6430 U.S.A.; (2) department of Plant Sciences, Downing Site, University of Cambridge, Cambridge, CB2 3EA, United Kingdom. MPMI 8:610-620. Accepted 2 March 1995. Copyright 1995 The American Phytopathological Society.

Syringomycin is a necrosis-inducing lipopeptide toxin synthesized and secreted by the phytopathogen, Pseudomonas syringae pv. syringae. Although small quantities of syringomycin are known to activate a cascade of physiological events in plasma membranes, the mechanism of action of the phytotoxin has never been fully characterized. The objective of this study was to test the hypothesis that the primary mode of action of syringomycin is to form transmembrane pores that are permeable to cations. Accordingly, direct measurement of ion fluxes were performed using artificial bilayers. The hemolytic properties and surface activity of HPLC-purified syringomycin were quanlified by use of an erythrocyte lysis assay and by the drop weight method. Assays were performed using syringomycin form SRE alone or a mixture containing all forms of the phytotoxin. At a threshold concentration of 500 ng/ml, syringomycin induced hemolysis by forming ion channels in membranes. Osmotic protection studies indicated a channel radius of between 0.6 and 1 nm. The ion channel-forming activity was insensitive and permeable to both monovalent and divalent cations, suggesting that syringomycin causes lysis of erythrocytes by colloid osmotic lysis. In addition, syringomycin, like other lipopeptide antibiotics, is a potent biosurfactant capable of lowering the interfacial tension of water to 31 mN/m. The critical micellar concentration of syringomycin was calculated to be 1.25 mg/ml and the (CMC was 33 mN/m. A model is presented depicting the mechanism of action of syringomycin in the plant-pathogen interaction. The model integrates known effects of the toxin on ion flux in plasma membranes with formation of ion channels and the consequential cascade of effects associated with cellular signalling.