Heterologous Expression Screens in <i>Nicotiana benthamiana</i> Identify a Candidate Effector of the Wheat Yellow Rust Pathogen that Associates with Processing Bodies

<div><p>Rust fungal pathogens of wheat (<i>Triticum</i> spp.) affect crop yields worldwide. The molecular mechanisms underlying the virulence of these pathogens remain elusive, due to the limited availability of suitable molecular genetic research tools. Notably, the inability to perform high-throughput analyses of candidate virulence proteins (also known as effectors) impairs progress. We previously established a pipeline for the fast-forward screens of rust fungal candidate effectors in the model plant <i>Nicotiana benthamiana</i>. This pipeline involves selecting candidate effectors <i>in silico</i> and performing cell biology and protein-protein interaction assays <i>in planta</i> to gain insight into the putative functions of candidate effectors. In this study, we used this pipeline to identify and characterize sixteen candidate effectors from the wheat yellow rust fungal pathogen <i>Puccinia striiformis</i> f sp <i>tritici</i>. Nine candidate effectors targeted a specific plant subcellular compartment or protein complex, providing valuable information on their putative functions in plant cells. One candidate effector, PST02549, accumulated in processing bodies (P-bodies), protein complexes involved in mRNA decapping, degradation, and storage. PST02549 also associates with the P-body-resident ENHANCER OF mRNA DECAPPING PROTEIN 4 (EDC4) from <i>N</i>. <i>benthamiana</i> and wheat. We propose that P-bodies are a novel plant cell compartment targeted by pathogen effectors.</p>