Methods to study PAMP-triggered immunity in Brassica species

Lloyd, Simon R, Schoonbeek, Henk-Jan, Trick, Martin, Zipfel, Cyril and Ridout, Christopher J (2014) Methods to study PAMP-triggered immunity in Brassica species. Molecular Plant-Microbe Interactions, 27 (3). pp. 286-295. ISSN 0894-0282

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Abstract

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.

Item Type: Article
Uncontrolled Keywords: amino acid sequence,botrytis,brassica napus,cell wall,gene expression profiling,gene expression regulation, plant,high-throughput nucleotide sequencing,mitogen-activated protein kinases,molecular sequence data,peptides,phosphorylation,plant diseases,plant immunity,plant leaves,plant proteins,pseudomonas syringae,rna, plant,reactive oxygen species,respiratory burst,seedling,sequence analysis, rna,species specificity
Faculty \ School: Faculty of Science > School of Biological Sciences
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Depositing User: Pure Connector
Date Deposited: 04 Jul 2014 12:43
Last Modified: 21 Apr 2020 23:06
URI: https://ueaeprints.uea.ac.uk/id/eprint/48863
DOI: 10.1094/MPMI-05-13-0154-FI

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