Specific binding of a hypersensitive lignification elicitor from Puccinia graminis f. sp. tritici to the plasma membrane from wheat (Triticum aestivum L.)

We have recently reported the isolation and characterization of a glycoprotein (M_r 67 000) from germ-tube walls of Puccinia graminis f. sp. tritici which elicits the cellular hypersensitive lignification response in wheat (G. Kogel et al., 1988, Physiol. Mol. Plant Pathol. 33, 173–185). The present study uses this glycoprotein, referred to as Pgt elicitor, to identify putative elicitor targets in wheat cell membranes. In enzyme-linked immunosorbent assays using anti-Pgt elicitor antibodies, specific binding sites for Pgt elicitor were detected in highly purified plasma-membrane vesicles of wheat (Triticum aestivum L.) primary leaf cells. Binding proved to be independent of the presence or absence in wheat of the Sr5 gene for rust resistance, and also occurred on barley (Hordeum vulgare L.) plasma membrane. The binding sites have an Mr of 30 000 and 33 000, respectively, and binding activity was not lost in the presence of sodium dodecyl sulfate. [¹⁴C]imido-Pgt elicitor was used to determine the apparent K_d value for specific binding, found to be 2.0 M, and the maximum content of binding sites, found to be 250 pmol per mg of plasma-membrane protein. The relevance of the elicitor binding for the outcome of the interaction of P. graminis and wheat is discussed.Abbreviations BSA bovine serum albumin - ELISA enzyme linked immunosorbent assay - IDPase inosine 5-diphosphatase - MPLC medium-pressure liquid chromatography - MF microsomal fraction - Pgt elicitor elicitor of Puccinia graminis f. sp. tritici - SDS sodium dodecyl sulfate - Pre U₃, Pre U₁ pure plasma membrane from wheat cultivar Prelude and plasma membrane contaminated by intracellular membrane, respectivelyThis work was supported by the Deutsche Forschungsgemeinschaft. We wish to thank C. Larsson, Lund, Sweden for his kind support in the preparation of plasma membrane.