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Methods to Study PAMP-Triggered Immunity in Brassica Species

March 2014 , Volume 27 , Number  3
Pages  286 - 295

Simon R. Lloyd,1 Henk-jan Schoonbeek,1 Martin Trick,2 Cyril Zipfel,3 and Christopher J. Ridout1

1Crop Genetics Department and 2Computational and Systems Biology Department, John Innes Centre, Norwich Research Park, Norwich, NR4 7UH, U.K.; 3The Sainsbury Laboratory, Norwich Research Park, Norwich, NR4 7UH, U.K.

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Accepted 6 October 2013.

The first layer of active defense in plants is based on the perception of pathogen-associated molecular patterns (PAMPs) leading to PAMP-triggered immunity (PTI). PTI is increasingly being investigated in crop plants, where it may have potential to provide durable disease resistance in the field. Limiting this work, however, is an absence of reliable bioassays to investigate PAMP responses in some species. Here, we present a series of methods to investigate PTI in Brassica napus. The assays allow measuring early responses such as the oxidative burst, mitogen-activated protein kinase phosphorylation, and PAMP-induced marker gene expression. Illumina-based RNA sequencing analysis produced a genome-wide survey of transcriptional changes upon PAMP treatment seen in both the A and C genomes of the allotetraploid B. napus. Later responses characterized include callose deposition and lignification at the cell wall, seedling growth inhibition, and PAMP-induced resistance to Pseudomonas syringae and Botrytis cinerea. Furthermore, using these assays, we demonstrated substantial variation in PAMP responses within a collection of diverse B. napus cultivars. The assays reported here could have widespread application in B. napus breeding and mapping programs to improve selection for broad-spectrum disease resistance.

© 2014 The American Phytopathological Society