In planta effector competition assays detect Hyaloperonospora arabidopsidis effectors that contribute to virulence and localize to different plant subcellular compartments

Badel, Jorge Luis, Piquerez, Sophie J M, Greenshields, David, Rallapalli, Ghanasyam, Fabro, Georgina, Ishaque, Naveed and Jones, Jonathan D G (2013) In planta effector competition assays detect Hyaloperonospora arabidopsidis effectors that contribute to virulence and localize to different plant subcellular compartments. Molecular Plant-Microbe Interactions, 26 (7). pp. 745-757. ISSN 0894-0282

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Abstract

The genome of the pathogenic oomycete Hyaloperonospora arabidopsidis is predicted to encode at least 134 high-confidence effectors (HaRxL) carrying the RxLR motif implicated in their translocation into plant cells. However, only four avirulence genes (ATR1, ATR13, ATR5, and ATR39) have been isolated. This indicates that identification of HaRxL effectors based on avirulence is low throughput. We aimed at rapidly identifying H. arabidopsidis effectors that contribute to virulence by developing methods to detect and quantify multiple candidates in bacterial mixed infections using either Illumina sequencing or capillary electrophoresis. In these assays, referred to here as in planta effector competition assays, we estimate the contribution to virulence of individual effectors by calculating the abundance of each HaRxL in the bacterial population recovered from leaves 3 days after inoculation relative to abundance in the initial mixed inoculum. We identified HaRxL that enhance Pseudomonas syringae pv. tomato DC3000 growth in some but not all Arabidopsis accessions. Further analysis showed that HaRxLL464, HaRxL75, HaRxL22, HaRxLL441, and HaRxL89 suppress pathogen-associated molecular pattern-triggered immunity (PTI) and localize to different subcellular compartments in Nicotiana benthamiana, providing evidence for a multilayered suppression of PTI by pathogenic oomycetes and molecular probes for the dissection of PTI.

Item Type: Article
Uncontrolled Keywords: amino acid motifs,antibiosis,arabidopsis,electrophoresis, capillary,high-throughput nucleotide sequencing,host-pathogen interactions,oomycetes,plant diseases,plant leaves,protein transport,proteins,pseudomonas syringae,recombinant fusion proteins,sequence analysis, dna,tobacco,virulence
Faculty \ School: Faculty of Medicine and Health Sciences > Norwich Medical School
Faculty of Science > School of Biological Sciences
University of East Anglia > Faculty of Science > Research Groups > Plant Sciences
Depositing User: Pure Connector
Date Deposited: 10 Nov 2016 11:00
Last Modified: 25 Jul 2018 12:54
URI: https://ueaeprints.uea.ac.uk/id/eprint/61303
DOI: 10.1094/MPMI-06-12-0154-R

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