A Wheat Protoplast Assay for Positive Effector Screening and Investigation of Host-Pathogen Interactions.

Abstract

Fungal pathogens present a severe risk to food systems; however, complex crop-microbe interactions are challenging to study using tools developed for model species. In particular, efficient screening and rapid assessment of microbial effectors is hindered by a lack of cloned resistance (R) genes and difficulty in validating large numbers of predicted effector candidates. This chapter describes a protocol for preparing wheat protoplasts to enable positive identification of host defense induction without overexpression of a cloned R gene, increasing the available pool of host resistance genes for screening. The assay uses polyethylene glycol (PEG)-calcium-mediated transient transfection to introduce candidate effector gene constructs into wheat protoplasts, with a defense-activated reporter for inducing a positive readout with internal normalization, indicating host recognition. This protocol provides a valuable tool for the study of host-pathogen interactions in wheat, contributing to improved resources for the development of disease-resistant crops and genome-informed pathogen surveillance.

Plant phenotyping relevance

コムギプロトプラストで宿主防御誘導を定量的に読み出すスクリーニング assay のプロトコル開発が中心であり、植物の防御状態を表現型として取得する方法に該当する。

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