Changes in the plasma membrane distribution of rice phospholipase D during resistant interactions with Xanthomonas oryzae pv oryzae

Abstract

Phospholipase D (PLD; EC 3.1.4.4), which hydrolyzes phospholipids to generate phosphatidic acid, was examined in rice leaves undergoing susceptible or resistant interactions with Xanthomonas oryzae pv oryzae. RNA analysis of leaves undergoing resistant interactions revealed different expression patterns for PLD over 5 days relative to control plants or those undergoing susceptible interactions. By using an activity assay and immunoblot analysis, we identified three forms of PLD (1, 2, and 3). PLD 1 was observed only at 1 day after tissue infiltration. PLDs 2 and 3 were detected up to 3 days in all interactions. Immunoelectron microscopy studies revealed PLD to be associated predominantly with the plasma membrane. In cells undergoing a susceptible response, PLD was uniformly distributed along the plasma membrane at 3, 6, 12, and 24 hr after inoculation. However, within 12 hr after bacterial challenge in resistant interactions, PLD was clustered preferentially in membranes adjacent to bacterial cells.