Abstract

Pyriculol was isolated from the rice blast fungus Magnaporthe oryzae and found to induce lesion formation on rice leaves. These findings suggest that it could be involved in virulence. The gene MoPKS19 was identified to encode a polyketide synthase essential for the production of the polyketide pyriculol in the rice blast fungus M. oryzae. The transcript abundance of MoPKS19 correlates with the biosynthesis rate of pyriculol in a time-dependent manner. Furthermore, gene inactivation of MoPKS19 resulted in a mutant unable to produce pyriculol, pyriculariol and their dihydro derivatives. Inactivation of a putative oxidase-encoding gene MoC19OXR1, which was found to be located in the genome close to MoPKS19, resulted in a mutant exclusively producing dihydropyriculol and dihydropyriculariol. By contrast, overexpression of MoC19OXR1 resulted in a mutant strain only producing pyriculol. The MoPKS19 cluster, furthermore, comprises two transcription factors MoC19TRF1 and MoC19TRF2, which were both found individually to act as negative regulators repressing gene expression of MoPKS19. Additionally, extracts of ΔMopks19 and ΔMoC19oxr1 made from axenic cultures failed to induce lesions on rice leaves compared to extracts of the wild-type strain. Consequently, pyriculol and its isomer pyriculariol appear to be the only lesion-inducing secondary metabolites produced by M. oryzae wild-type (MoWT) under these culture conditions. Interestingly, the mutants unable to produce pyriculol and pyriculariol were as pathogenic as MoWT, demonstrating that pyriculol is not required for infection.

Highlights

  • Fungal secondary metabolites play crucial roles in plant– pathogen interactions and it appears to be of particular interest to elucidate their function in respect of the biological activity and the organization of their biosynthetic pathways

  • In order to investigate the secondary metabolism of M. oryzae 70-15 (MoWT), the fungal strain was grown under different culture conditions that affect secondary metabolite production

  • Crude extracts of the submerged cultures were analysed by HPLC/MS to identify secondary metabolites produced by the rice blast fungus

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Summary

Introduction

Fungal secondary metabolites play crucial roles in plant– pathogen interactions and it appears to be of particular interest to elucidate their function in respect of the biological activity and the organization of their biosynthetic pathways. Magnaporthe oryzae causes rice blast disease and is one of the most significant plant pathogens worldwide [2]. This filamentous ascomycete has been studied intensively and has become an excellent model organism for studying the molecular mechanisms of plant–pathogen interaction. One of the fungal secondary metabolites studied most intensively is the polyketide dihydroxynaphthalene (DHN) melanin. Further secondary metabolites involved in the virulence of the rice blast fungus were believed to be produced by the polyketide synthase [PKS-nonribosomal peptide synthase (NRPS)] MoAce1p. The gene MoACE1 encodes an intracellular hybrid protein which was found to be involved in effects concerning the rice resistance gene Pi33 [7, 8]

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