Abstract

The rice blast fungus Magnaporthe oryzae causes plant disease via specialised infection structures called appressoria. These dome-shaped cells are able to generate enormous internal pressure, which enables penetration of rice tissue by invasive hyphae. Previous studies have shown that mobilisation of lipid bodies and subsequent lipid metabolism are essential pre-requisites for successful appressorium-mediated plant infection, which requires autophagic recycling of the contents of germinated spores and germ tubes to the developing appressorium. Here, we set out to identify putative regulators of lipid metabolism in the rice blast fungus. We report the identification of FAR1 and FAR2, which encode highly conserved members of the Zn2-Cys6 family of transcriptional regulators. We generated Δfar1, Δfar2 and Δfar1Δfar2 double mutants in M. oryzae and show that these deletion mutants are deficient in growth on long chain fatty acids. In addition, Δfar2 mutants are also unable to grow on acetate and short chain fatty acids. FAR1 and FAR2 are necessary for differential expression of genes involved in fatty acid β-oxidation, acetyl-CoA translocation, peroxisomal biogenesis, and the glyoxylate cycle in response to the presence of lipids. Furthermore, FAR2 is necessary for expression of genes associated with acetyl-CoA synthesis. Interestingly, Δfar1, Δfar2 and Δfar1Δfar2 mutants show no observable delay or reduction in lipid body mobilisation during plant infection, suggesting that these transcriptional regulators control lipid substrate utilization by the fungus but not the mobilisation of intracellular lipid reserves during infection-related morphogenesis.

Highlights

  • Rice blast disease is caused by the fungus Magnaporthe oryzae and is one of the most destructive diseases of cultivated rice

  • We demonstrate that FAR1 and FAR2 are necessary for the differential expression of genes involved in fatty acid b-oxidation, acetyl-coenzyme A (CoA) translocation, peroxisome biogenesis, and the glyoxylate cycle in response to the presence of lipids

  • Lipolysis in the appressorium leads to b-oxidation of fatty acids, generating acetyl CoA, which is metabolized via the glyoxylate shunt bypass enabling gluconeogenesis [11,12,13,14], but is a metabolic intermediate necessary for synthesis of melanin, isoprenoids, amino acids, and a range of secondary metabolites

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Summary

Introduction

Rice blast disease is caused by the fungus Magnaporthe oryzae and is one of the most destructive diseases of cultivated rice. Serious harvest losses can occur in all rice-growing regions of the world, with up to 18% yield losses per annum [1]. The spread of rice blast disease occurs by dispersal of asexual spores, called conidia, which adhere strongly to the leaf surface. A conidium germinates on the leaf cuticle and develops a polarised germ tube, which swells at its tip to form a single-celled appressorium. The appressorium accumulates high concentrations of osmotically compatible solutes, including glycerol, which generates huge internal pressure that the fungus utilises to force a narrow penetration hypha through the plant cuticle and into the leaf epidermis [2]

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