In-depth insight into in vivo apoplastic secretome of rice-Magnaporthe oryzae interaction

Abstract

The in vivo apoplastic fluid secretome of rice-blast fungus interaction remains largely uncharacterized. Here, we report a proteomics investigation of in vivo secreted proteins of rice leaves infected with incompatible (KJ401) and compatible (KJ301) races of Magnaporthe oryzae (M. oryzae) using 2-DGE and MudPIT coupled with MALDI-TOF-MS and/or nESI-LC-MS/MS analyses. Prepared fractions of secretory proteins were essentially free from cytoplasmic contamination. Two-DGE and MudPIT identified 732 secretory proteins, where 291 (40%) and 441 (60%) proteins were derived from rice and M. oryzae, respectively. Of these, 39.2% (rice) and 38.9% (M. oryzae) of proteins were predicted by SignalP as retaining signal peptides. Among these, rice secreted more proteins related to stress response, ROS and energy metabolism, whereas, M. oryzae secreted more proteins involved in metabolism and cell wall hydrolyses. Semi-quantitative RT-PCR revealed their differential expression under compatible/incompatible interactions. In vivo expression of M. oryzae glycosyl hydrolase (GH) protein family members using particle bombardment driven transient expression system showed that four GH genes could act as effectors within host apoplast possibly via interaction with host membrane bound receptor. The established in vivo secretome serves as a valuable resource toward secretome analysis of rice-M. oryzae interaction. (C) 2012 Elsevier B.V. All rights reserved.