Open AccessRapid Methodologies for AssessingPseudomonas syringaepv.actinidiaeColonization and Effector-Mediated Hypersensitive Response in Kiwifruit
Author(s) -
Jay Jayaraman,
Abhishek Chatterjee,
Shan Hunter,
Ronan Chen,
Erin A. Stroud,
Hassan Saei,
Stephen Hoyte,
Simon C. Deroles,
Jibran Tahir,
Matthew D. Templeton,
Cyril Brendolise
Publication year - 2021
Publication title -
molecular plant-microbe interactions
Language(s) - English
Resource type - Journals
eISSN - 1943-7706
pISSN - 0894-0282
DOI - 10.1094/mpmi-02-21-0043-r
Subject(s) - pseudomonas syringae , biology , effector , pseudomonas fluorescens , pathogen , gene , microbiology and biotechnology , computational biology , genetics , bacteria
The infection of Pseudomonas syringae pv. actinidiae in kiwifruit is currently assessed by numerous methodologies, each with their own limitations. Most studies are based on either a laborious method of growth quantification of the pathogen or qualitative assessments by visual scoring following stem or cutting inoculation. Additionally, when assessing for resistance against specific pathogen effectors, confounding interactions between multiple genes in the pathogen can make mapping resistance phenotypes nearly impossible. Here, we present robust alternative methods to quantify pathogen load based on rapid bacterial DNA quantification by PCR, the use of Pseudomonas fluorescens, and a transient reporter eclipse assay for assessing resistance conferred by isolated bacterial avirulence genes. These assays compare well with bacterial plate counts to assess bacterial colonization as a result of plant resistance activation. The DNA-based quantification, when coupled with the P. fluorescens and reporter eclipse assays to independently identify bacterial avirulence genes, is rapid, highly reproducible, and scalable for high-throughput screens of multiple cultivars or genotypes. Application of these methodologies will allow rapid and high-throughput identification of resistant cultivars and the bacterial avirulence genes they recognize, facilitating resistance gene discovery for plant breeding programs. [Formula: see text] Copyright © 2021 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .