Role for RNA-Binding Proteins Implicated in Pathogenic Development of Ustilago maydis
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ABSTRACT Ustilago maydis causes smut disease on corn. Successful infection depends on a number of morphological transitions, such as pheromone-dependent formation of conjugation tubes and the switch to filamentous dikaryotic growth, as well as different types of mycelial structures during growth within the host plant. In order to address the involvement of RNA-binding proteins during this developmental program, we identified 27 open reading frames from the genome sequence encoding potential RNA-binding proteins. They exhibit similarities to RNA-binding proteins with Pumilio homology domains (PUM), the K homology domain (KHD), the double-stranded RNA binding motif (DSRM), and the RNA recognition motif (RRM). For 18 of these genes, we generated replacement mutants in compatible haploid strains. Through analysis of growth behavior, morphology, cyclic AMP response, mating, and pathogenicity, we identified three candidates with aberrant phenotypes. Loss of Khd1, a K homology protein containing three KHDs, resulted in a cold-sensitive growth phenotype. Deletion of khd4 encoding a protein with five KHDs led to abnormal cell morphology, reduced mating, and virulence. rrm4 Δ strains were affected in filamentous growth and pathogenicity. Rrm4 is an RRM protein with a so far unique domain organization consisting of three N-terminal RRMs as well as a domain found in the C terminus of poly(A)-binding proteins. These results indicate a role for RNA-binding proteins in regulation of morphology as well as in pathogenic development in U. maydis .Keywords:
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Polyamines are implicated in the regulation of many processes in the plant cell, including functioning of ion channels, DNA replication, gene transcription, mRNA translation, cell proliferation and programmed cell death. Plant polyamines occur either in free form, covalently bound to proteins, or conjugated to hydroxycinnamic acids forming phenol amides. Ustilago maydis is a dimorphic and biotrophic pathogenic fungus responsible for common smut or ‘‘huitlacoche’’ in maize; and it is considered an excellent model for the study of plant-pathogen interactions. Recently, we reported alterations in polyamine metabolism of maize tumors induced on leaf blades by Ustilago maydis infection. Our data revealed a striking increase in maize polyamine biosynthesis, mainly free and conjugated putrescine in the tumors and in the green plant tissue surrounding the tumor. In this addendum, we describe that changes in PA metabolism take place even in earlier stages of maize plant infection with Ustilago maydis.
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Abstract In the plant Ustilago maydis causes: 1. A lowering of free radical concentrations in tissues of nodal roots and in internodial tissues close to the fungus. 2. Conversely, in tissues of the primary root, of aerial roots, of leaves and nodes 1–5, 13, 15 and 16 it causes a rise of free radicals concentration. Ustilago maydis has a similar effect upon free radicals concentration in maize leaves as Erysiphe graminis in barley leaves.
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Abstract The Isolation of Soil‐Inhabiting Enemies of Ustilago maydis Using sporidial lawns of Ustilago maydis in water agar layers as indicator plates 2 organisms could be isolated from soil samples which produce plaques. These organisms were identified as Acanthamoeba polyphaga and Myxococcus spp. Both organisms control populations of Ustilago maydis in sterile soil too. The isolation method may be gainfully used in the search for further organisms with more specific potential for the biological control of Ustilago maydis .
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Ustilago maydis is a phytopathogenic fungus responsible for corn smut disease. Although it is a very well-established model organism for the study of plant-microbe interactions, its potential to produce specialized metabolites, which might contribute to this interaction, has not been studied in detail. By analyzing the U. maydis genome, we identified a biosynthetic gene cluster whose activation led to the production of a black melanin pigment. Single deletion mutants of the cluster genes revealed that five encoded enzymes are required for the accumulation of the black pigment, including three polyketide synthases (pks3, pks4, and pks5), a cytochrome P450 monooxygenase (cyp4), and a protein with similarity to versicolorin B synthase (vbs1). Metabolic profiles of deletion mutants in this gene cluster suggested that Pks3 and Pks4 act in concert as heterodimers to generate orsellinic acid (OA), which is reduced to the corresponding aldehyde by Pks5. The OA-aldehyde can then react with triacetic acid lactone (TAL), also derived from Pks3/Pks4 heterodimers to form larger molecules, including novel coumarin derivatives. Our findings suggest that U. maydis synthesizes a novel type of melanin based on coumarin and pyran-2-one intermediates, while most fungal melanins are derived from 1,8-dihydroxynaphthalene (DHN) or l-3,4-dihydroxyphenylalanine (l-DOPA). Along with these observations, this work also provides insight into the mechanisms of polyketide synthases in this filamentous fungus.IMPORTANCE The fungus Ustilago maydis represents one of the major threats to maize plants since it is responsible for corn smut disease, which generates considerable economical losses around the world. Therefore, contributing to a better understanding of the biochemistry of defense mechanisms used by U. maydis to protect itself against harsh environments, such as the synthesis of melanin, could provide improved biological tools for tackling the problem and protect the crops. In addition, the fact that this fungus synthesizes melanin in an unconventional way, requiring more than one polyketide synthase for producing melanin precursors, gives a different perspective on the complexity of these multidomain enzymes and their evolution in the fungal kingdom.
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Bacteria and fungi were tested for sensitivity to Ustilago maydis killer strains carrying virus-like particles. Various species taxonomically related to U. maydis were sensitive.
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