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Molecular & Cellular Proteomics 8:2368-2381, 2009.
© 2009 by The American Society for Biochemistry and Molecular Biology, Inc.

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Research

In Planta Proteomics and Proteogenomics of the Biotrophic Barley Fungal Pathogen Blumeria graminis f. sp. hordei^*,

Laurence V. Bindschedler^,, Timothy A. Burgis^¶, Davinia J. S. Mills, Jenny T. C. Ho^||, Rainer Cramer^,** and Pietro D. Spanu^,

From The BioCentre and
School of Biological Sciences, The University of Reading, P. O. Box 221, Reading RG6 6AS, United Kingdom,
¶Centre for Bioinformatics, Imperial College, Fleming Building, South Kensington Campus, London SW7 2AZ, United Kingdom,
||Thermo Fisher Scientific, Stafford House, Boundary Way, Hemel Hempstead, HP2 7GE, United Kingdom,
**Department of Chemistry, The University of Reading, P. O. Box 224, Reading RG6 6AD, United Kingdom, and
Department of Life Sciences, Imperial College London, South Kensington Campus, London SW7 2AZ, United Kingdom

To further our understanding of powdery mildew biology during infection, we undertook a systematic shotgun proteomics analysis of the obligate biotroph Blumeria graminis f. sp. hordei at different stages of development in the host. Moreover we used a proteogenomics approach to feed information into the annotation of the newly sequenced genome. We analyzed and compared the proteomes from three stages of development representing different functions during the plant-dependent vegetative life cycle of this fungus. We identified 441 proteins in ungerminated spores, 775 proteins in epiphytic sporulating hyphae, and 47 proteins from haustoria inside barley leaf epidermal cells and used the data to aid annotation of the B. graminis f. sp. hordei genome. We also compared the differences in the protein complement of these key stages. Although confirming some of the previously reported findings and models derived from the analysis of transcriptome dynamics, our results also suggest that the intracellular haustoria are subject to stress possibly as a result of the plant defense strategy, including the production of reactive oxygen species. In addition, a number of small haustorial proteins with a predicted N-terminal signal peptide for secretion were identified in infected tissues: these represent candidate effector proteins that may play a role in controlling host metabolism and immunity.

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