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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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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