Abstract
Interactions between plants and compatible fungal pathogens are spatially and temporally dynamic, posing a major challenge for sampling and data analysis. A protocol is described for the infection of the model grass species Brachypodium distachyon with Magnaporthe grisea (rice blast), together with modifications to extend the use to rice and barley. We outline a method for the preparation of long-term stocks of virulent fungal pathogens and for the generation of fungal inoculants for challenge of host plants. Host plant growth, pathogen inoculation and plant sampling protocols are presented together with methods for assessing the efficiency of both infection and sampling procedures. Included in the anticipated results is a description of the use of metabolite fingerprinting and multivariate data analysis to assess disease synchrony and validate system reproducibility between experiments. The design concepts will have value in any studies using biological systems that contain dynamic variance associated with large compositional changes in sample matrix over time.
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References
Xu, J.R. et al. The dawn of fungal pathogen genomics. Annu. Rev. Phytopathol. 44, 337–366 (2006).
Talbot, N.J. On the trail of a cereal killer: exploring the biology of Magnaporthe grisea. Annu. Rev. Micro. 57, 177–202 (2003).
Goswani, R.S. & Kistler, H.C. Heading for disaster: Fusarium graminearum on cereal crops. Mol. Plant. Pathol. 5, 515–525 (2004).
Basse, C.W. & Steinberg, G. Ustilago maydis, model system for analysis of the molecular basis of fungal pathogenicity. Mol. Plant Pathol. 5, 83–92 (2004).
Howard, R.J. & Valent, B. Breaking and entering: host penetration by the fungal rice blast pathogen Magnaporthe grisea. Annu. Rev. Microbiol. 50, 491–512 (1996).
Chisholm, S.T. et al. Host-microbe interactions: shaping the evolution of the plant immune response. Cell 124, 803–814 (2006).
Martin, G.B. et al. Understanding the functions of plant disease resistance proteins. Annu. Rev. Plant Biol. 54, 23–61 (2003).
Greenberg, J.T. & Yao, N. The role and regulation of programmed cell death in plant-pathogen interactions. Cell Microbiol. 6, 201–211 (2004).
Scholes, J., Lee, P., Horton, P. & Lewis, D. Invertase: understanding changes in the photosynthetic and carbohydrate metabolism of barley leaves infected with powdery mildew. New Phytol. 126, 213–222 (1994).
Swarbrick, P., Schulze-Lefert, P. & Scholes, J. Metabolic consequences of susceptibility and resistance (race-specific and broad spectrum) in barley leaves challenged with powdery mildew. Plant Cell. Environ. 29, 1061–1076 (2006).
Nomura, K. et al. Suppression of host defence in compatible plant-Pseudomonas syringae interactions. Curr. Opin. Plant Biol. 8, 361–368 (2005).
Solomon, P.S., Tan, K-C. & Oliver, R.P. The nutrient supply of pathogenic fungi; a fertile field for study. Mol. Plant Patholol. 4, 203–210 (2003).
Kamoun, S. A catalogue of the effector secretome of plant pathogenic oomycetes. Ann. Rev. Phytopathol. 44, 41–60 (2006).
Draper, J. et al. Brachypodium distachyon. A new model system for functional genomics in grasses. Plant Physiol. 127, 1539–1555 (2001).
Cousin, A. et al. The MAP kinase-encoding gene MgFus3 of the non-appressorium phytopathogen Mycosphaerella graminicola is required for penetration and in vitro pycnidia formation. Mol. Plant Pathol. 7, 269–278 (2006).
Slawecki, R.A. et al. Novel fungitoxicity assays for inhibition of germination-associated adhesion of Botrytis cinerea and Puccinia recondita spores. Appl. Envirol. Microbiol. 68, 597–601 (2002).
Schurch, S. et al. Molecular population genetic analysis differentiates two virulence mechanisms of the fungal avirulence gene NIP1. Mol. Plant Microbe Interact. 17, 1114–1125 (2004).
Jones, M.J. & Dunkle, L.D. Factors regulating the synthesis of a cyclic peptide pathotoxin produced by Cochliobolus carbonum. Mycol. Res. 102, 1381–1386 (1998).
Dean, R.A. et al. The genome sequence of the rice blast fungus Magnaporthe grisea. Nature 434, 980–986 (2005).
Sesma, A. & Osbourn, A.E. The rice leaf blast pathogen undergoes developmental processes typical of root-infecting fungi. Nature 431, 582–586 (2004).
Ou, S.H. Rice diseases. Commonwealth Mycological Institute Kew Surrey (1987).
Routledge, A. et al. Magnaporthe grisea interactions with the model grass Brachypodium distachyon closely resemble those with rice (Oryza sativa). Mol. Plant Pathol. 5, 253–265 (2004).
Talbot, N.J., Saich, Y.P., Ma, M. & Hamer, J.E. Karyotypic variation within clonal lineages of the rice blast fungus, Magnaporthe grisea. Appl. Environ. Microbiol. 59, 585–593 (1993).
Soanes, D.M. et al. Regulation of the MPG1 hydrophobin gene in the rice blast fungus Magnaporthe grisea. Mol. Plant Microbe Interact. 15, 1253–1267 (2002).
Giles, P.F. et al. A relational database for the discovery of genes encoding amino acid biosynthetic enzymes in pathogenic fungi. Comp. Funct. Genom. 4, 4–15 (2003).
Hamer, J.E. et al. A mechanism for surface attachment in spores of a plant pathogenic fungus. Science 239, 288–290 (1988).
Valent, B. & Chumley, F.G. Molecular genetic-analysis of the rice blast fungus, Magnaporthe grisea. Annu. Rev. Phytopathol. 29, 443–467 (1991).
Veneault-Fourrey, C. et al. Fungal cell death is necessary for infection by the rice blast fungus. Science 312, 580–583 (2006).
Kankanala, P., Czymmek, K. & Valent, B. Roles for rice membrane dynamics and plasmodesmata during biotrophic invasion by the blast fungus. Plant Cell 19, 706–724 (2007).
Talbot, N. Having a blast: exploring the pathogenicity of Magnaporthe grisea. Trends Microbiol. 3, 9–16 (1995).
Perfect, S.E. & Green, J.R. Infection structures of biotrophic and hemibiotrophic fungal plant pathogens. Mol. Plant Pathol. 2, 101–108 (2001).
Hood, M.E. & Shew, H.D. Application of KOH-analine blue fluorescence in the study of plant-fungal interactions. Phytopathol. 86, 704–708 (1996).
Beckmann, M., Parker, D., Enot, D.P., Duval, E. & Draper, J. High-throughput, nontargeted metabolite fingerprinting using nominal mass flow injection electrospray mass spectrometry. Nat. Protoc. 3, 486–504 (2008).
Enot, D.P. et al. Preprocessing, classification modeling and feature selection using flow injection electrospray mass spectrometry metabolite fingerprint data. Nat. Protoc. 3, 446–470 (2008).
Acknowledgements
We thank all collaborators who provided valuable samples to develop this protocol and in particular Rob Darby for maintaining laboratory infrastructure, equipment and materials. Metabolite analysis, statistical work and biological materials used in example data were generated as part of the UK Biotechnology and Biological Sciences Research Council grants BB/D006953/1 (D.P. and J.D.), BB/D006791/1 (N.T. and Z.C.R.).
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Parker, D., Beckmann, M., Enot, D. et al. Rice blast infection of Brachypodium distachyon as a model system to study dynamic host/pathogen interactions. Nat Protoc 3, 435–445 (2008). https://doi.org/10.1038/nprot.2007.499
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DOI: https://doi.org/10.1038/nprot.2007.499
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