Garnica, Diana PUpadhyaya, Narayana MDodds, Peter NRathjen, John P2015-11-252015-11-251932-6203http://hdl.handle.net/1885/16744Stripe rust caused by the fungus Puccinia striiformis f.sp. tritici (Pst) is a major constraint to wheat production worldwide. The molecular events that underlie Pst pathogenicity are largely unknown. Like all rusts, Pst creates a specialized cellular structure within host cells called the haustorium to obtain nutrients from wheat, and to secrete pathogenicity factors called effector proteins. We purified Pst haustoria and used next-generation sequencing platforms to assemble the haustorial transcriptome as well as the transcriptome of germinated spores. 12,282 transcripts were assembled from 454-pyrosequencing data and used as reference for digital gene expression analysis to compare the germinated uredinospores and haustoria transcriptomes based on Illumina RNAseq data. More than 400 genes encoding secreted proteins which constitute candidate effectors were identified from the haustorial transcriptome, with two thirds of these up-regulated in this tissue compared to germinated spores. RT-PCR analysis confirmed the expression patterns of 94 effector candidates. The analysis also revealed that spores rely mainly on stored energy reserves for growth and development, while haustoria take up host nutrients for massive energy production for biosynthetic pathways and the ultimate production of spores. Together, these studies substantially increase our knowledge of potential Pst effectors and provide new insights into the pathogenic strategies of this important organism.J.P.R. is an ARC Future Fellow (FT0992129). This project has been supported by Bioplatforms Australia through funding from the Commonwealth Government NCRIS and Education Investment Fund Super Science programs.17 pages© 2013 Garnica et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.stripe rustPuccinia striiformis f.sp. tritici (Pst)wheat productionhaustoriumpathogenicity factorseffector proteinsnext-generation sequencing platformshaustorial transcriptometranscriptome of germinated spores2013-06-2610.1371/journal.pone.00671502015-12-11