Abstract

Grey mould is an important necrotrophic fungal pathogen that causes huge economic losses in agriculture. Many types of bacteria are used for biological control of grey mould via competition for space or nutrients and/or the production of antifungal metabolites. Oxalate is a key component of virulent necrotic fungal pathogens. In this study, we isolated non-antifungal oxalate-degrading bacteria (ODB) from the surfaces of oxalate-rich spinach and strawberries to investigate their ability to control necrotic fungal pathogens such as grey mould. A total of 36 bacteria grown on oxalate minimal (OM) agar plates were tested for oxalate-degrading activity. Five isolates exhibiting the highest oxalate degradation activity were subjected to molecular identification using 16S rRNA gene sequencing. Two isolates exhibiting non-antifungal activity were subjected to disease suppression assays using Arabidopsis–Botrytis systems. The isolate Pseudomonas abietaniphila ODB36, which exhibited significant plant protective ability, was finally selected for further investigation. Based on whole-genome information, the pseudomonad oxalate degrading (podA) gene, which encodes formyl-CoA transferase, was analysed. The podA− mutant did not inhibit Botrytis infection and oxalate toxicity; the defects were recovered by podA complementation. Purified PodA–His converted oxalate to formate and eliminated oxalate toxicity. These results indicate that P. abietaniphila ODB36 and PodA enzyme are associated with various aspects of grey mould disease inhibitory effects.

Highlights

  • Botrytis cinerea, which causes grey mould, is an important necrotrophic pathogen that causes huge economic losses in agriculture[1]

  • A total of 36 bacteria grown on oxalate minimal (OM)

  • The oxalate-degrading activity of these 36 bacteria was tested in OM broth supplemented with 1/10 Bacto tryptic soy broth (TSB)

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Summary

Introduction

Botrytis cinerea, which causes grey mould, is an important necrotrophic pathogen that causes huge economic losses in agriculture[1]. The modes of antagonistic bacterial activity include competition, antibiosis, lytic enzyme production, interference with pathogen activity and growth, volatility, and host resistance induction. Many necrotrophic pathogens including grey mould are known to infect various host plants via the production of oxalate, which is toxic to cellular organisms including mammals and plants[8,9]. Oxalate acts as a strong chelator of cations; it greatly oxidizes organic compounds in necrotrophic pathogens[10], helps to reduce pH, thereby facilitating plant infection by pectolytic enzymes, and impedes defense signaling pathways in the host[11]. The oxalate-degrading bacterial strain Cupriavidus campinensis was reported to protect Arabidopsis thaliana and crop species against B. cinerea[13]

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