Suppression of Chitin-Triggered Immunity by a New Fungal Chitin-Binding Effector Resulting from Alternative Splicing of a Chitin Deacetylase Gene

Abstract

Phytopathogenic fungi have evolved mechanisms to manipulate plant defences, such as chitin-triggered immunity, a plant defensive response based on the recognition of chitin oligomers by plant-specific receptors. To cope with chitin resistance, fungal pathogens have developed different strategies to prevent chitin recognition, such as binding, breaking, or modifying immunogenic oligomers. In powdery mildew fungi, the activity of chitin deacetylase (CDA) is crucial for this purpose, since silencing of the CDA gene leads to a rapid activation of chitin signalling and the subsequent suppression of fungal growth. In this work, we have identified an unusually short CDA transcript in Podosphaera xanthii, the cucurbit powdery mildew pathogen. This transcript, designated PxCDA3, appears to encode a truncated version of CDA resulting from an alternative splicing of the PxCDA gene, which lacked most of the chitin deacetylase activity domain but retained the carbohydrate-binding module. Experiments with the recombinant protein showed its ability to bind to chitin oligomers and prevent the activation of chitin signalling. Furthermore, the use of fluorescent fusion proteins allowed its localization in plant papillae at pathogen penetration sites. Our results suggest the occurrence of a new fungal chitin-binding effector, designated CHBE, involved in the manipulation of chitin-triggered immunity in powdery mildew fungi.