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Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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The WOPR protein Ros1 is a master regulator of sporogenesis and effector gene expression in the maize pathogen Ustilago maydis [RNA-Seq]

ABSTRACT: Goals: characterization of the transcription factor Ros1 in Ustilago maydis Methods: generation of deletion mutants, microscopic observations, ectopic expression of ros1, identification of Ros1 regulated genes by RNAseq and ChIP sequencing Results: Ros1 is not involved in plant colonization but is essential to trigger sporogenesis during late stages of infection. Premature expression of ros1 revealed that Ros1 counteracts the b-dependent filamentation program and induces morphological alterations resembling the early steps of sporogenesis. Transcriptional profiling and ChIP seq analyses revealed that Ros1 affects the expression of about 30 % of all U. maydis genes with 40% being direct targets. Cell wall remodeling and plasma membrane modifications are among the processes affected by Ros1 dependent regulation. Interestingly a large number of b-dependent genes including transcription factors and effector genes involved in biotrophy establishment were downregulated by Ros1 while a subset of novel â??late effectorsâ?? were upregulated. Taken together our results indicate that Ros1 is a master regulator of sporogenesis in U. maydis and that the switch to sporogenesis is accompanied by the differential regulation of 75% of the effector genes. Two samples corresponding to plant material infected with either U. maydis wild type strains FB1 x FB2 or the ros1 deletion strains FB1Dros1 x FB2Dros1 were analyzed in triplicate.

ORGANISM(S): Ustilago maydis

SUBMITTER: Marie Tollot

PROVIDER: E-GEOD-76229 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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The WOPR protein Ros1 is a master regulator of sporogenesis and effector gene expression in the maize pathogen Ustilago maydis [ChIP-Seq]

Project description:Goals: characterization of the transcription factor Ros1 in Ustilago maydis Methods: generation of deletion mutants, microscopic observations, ectopic expression of ros1, identification of Ros1 regulated genes by RNAseq and ChIP sequencing Results: Ros1 is not involved in plant colonization but is essential to trigger sporogenesis during late stages of infection. Premature expression of ros1 revealed that Ros1 counteracts the b-dependent filamentation program and induces morphological alterations resembling the early steps of sporogenesis. Transcriptional profiling and ChIP seq analyses revealed that Ros1 affects the expression of about 30 % of all U. maydis genes with 40% being direct targets. Cell wall remodeling and plasma membrane modifications are among the processes affected by Ros1 dependent regulation. Interestingly a large number of b-dependent genes including transcription factors and effector genes involved in biotrophy establishment were downregulated by Ros1 while a subset of novel â??late effectorsâ?? were upregulated. Taken together our results indicate that Ros1 is a master regulator of sporogenesis in U. maydis and that the switch to sporogenesis is accompanied by the differential regulation of 75% of the effector genes. 4 samples were analyzed in triplicate. The first two samples correspond to input and output DNA obtained from ChIP carried out on plant material infected with U. maydis strains expressing Ros1 with an HA tag. The other two other samples correspond to input and output DNA from ChIP carried out on plant material infected with U. maydis strains expressing Ros1 without a tag (control samples).

2016-06-24 | E-GEOD-76230 | biostudies-arrayexpress

The WOPR protein Ros1 is a master regulator of sporogenesis and effector gene expression in the maize pathogen Ustilago maydis [ChIP-Seq]

Project description:Goals: characterization of the transcription factor Ros1 in Ustilago maydis Methods: generation of deletion mutants, microscopic observations, ectopic expression of ros1, identification of Ros1 regulated genes by RNAseq and ChIP sequencing Results: Ros1 is not involved in plant colonization but is essential to trigger sporogenesis during late stages of infection. Premature expression of ros1 revealed that Ros1 counteracts the b-dependent filamentation program and induces morphological alterations resembling the early steps of sporogenesis. Transcriptional profiling and ChIP seq analyses revealed that Ros1 affects the expression of about 30 % of all U. maydis genes with 40% being direct targets. Cell wall remodeling and plasma membrane modifications are among the processes affected by Ros1 dependent regulation. Interestingly a large number of b-dependent genes including transcription factors and effector genes involved in biotrophy establishment were downregulated by Ros1 while a subset of novel “late effectors” were upregulated. Taken together our results indicate that Ros1 is a master regulator of sporogenesis in U. maydis and that the switch to sporogenesis is accompanied by the differential regulation of 75% of the effector genes.

2016-06-24 | GSE76230 | GEO

The WOPR protein Ros1 is a master regulator of sporogenesis and effector gene expression in the maize pathogen Ustilago maydis [RNA-Seq]

Project description:Goals: characterization of the transcription factor Ros1 in Ustilago maydis Methods: generation of deletion mutants, microscopic observations, ectopic expression of ros1, identification of Ros1 regulated genes by RNAseq and ChIP sequencing Results: Ros1 is not involved in plant colonization but is essential to trigger sporogenesis during late stages of infection. Premature expression of ros1 revealed that Ros1 counteracts the b-dependent filamentation program and induces morphological alterations resembling the early steps of sporogenesis. Transcriptional profiling and ChIP seq analyses revealed that Ros1 affects the expression of about 30 % of all U. maydis genes with 40% being direct targets. Cell wall remodeling and plasma membrane modifications are among the processes affected by Ros1 dependent regulation. Interestingly a large number of b-dependent genes including transcription factors and effector genes involved in biotrophy establishment were downregulated by Ros1 while a subset of novel “late effectors” were upregulated. Taken together our results indicate that Ros1 is a master regulator of sporogenesis in U. maydis and that the switch to sporogenesis is accompanied by the differential regulation of 75% of the effector genes.

2016-06-24 | GSE76229 | GEO

Transcriptome analysis of a Ustilago maydis ust1 deletion mutant

Project description:Ustilago maydis, the causal agent of corn smut disease, is a dimorphic fungus alternating between a saprobic haploid budding form, and an obligate pathogenic filamentous dikaryon. Maize responds to U. maydis colonization by producing highly modified tumorous structures and it is only within these plant galls that the fungus sporulates giving rise to melanized sexual spores, the teliospores. Previously we identified a regulatory protein from the APSES family of transcription factors, which we named Ust1, whose absence in yeast cells led to filamentous growth and the production of highly pigmented spore-like structures in culture. In this study, we analyzed the transcriptome of a ?ust1 deletion mutant. To identify genes potentially involved in the sporulation program, we carried out microarray analysis comparing a haploid U. maydis WT strain (1/2) and ust1 (14/25) in vitro. Wild-type and ?ust1 strains were grown in potato dextrose broth (PDB) for 48 h at 30oC. To allow effective comparison with other array data generated by U. maydis researchers, cells were then transferred to liquid array medium (6.25% Holiday salt solution, 30mM L-glutamine, 1% glucose, pH7.0, filter sterilized) at 30 C. Total RNA samples from WT 1/2 and ?ust1 mutant strains grown in array medium for 24 h (mutant’s filamentous phase), and 48 h (mutant’s spore-like structure formation phase) were extracted and purified (Sigma Spectrum Plant Total RNA Kit, Cat.No.STRN50). RNA was sent to NimbleGen, where it was reverse transcribed to cDNA and Cy3 labeled. The cDNA samples were hybridized, microarray chips scanned, and raw data normalized with appropriate controls.

2014-02-04 | E-GEOD-54659 | biostudies-arrayexpress

A secreted effector protein of Ustilago maydis guides maize leaf cells to form tumors

Project description:Zea mays transcriptome profiling of infected seedlings by the Ustilago maydis wildtype and the seedling specific effector mutant demonstrated the variation of gene expression in the mutant and the classes of genes that are absent in the mutant as compared to the wildtype U. maydis SG200 strain. Two dye competitive hybridizations were performed on Agilent Oligo arrays. Comparison were done 1) with mock and infected samples at 6dpi. 2) between the two 6dpi infected samples with wildtype and the secreted effector mutant

2015-03-09 | E-GEOD-63077 | biostudies-arrayexpress

Gene expression during biotrophic growth

Project description:Comparison between Ustilago maydis cells grown in axenic culture and U. maydis cells from plant tumors. Keywords: developmental stages U. maydis FB1 cells were grown in axenic culture; corn plants were infected with a mixture of strains FB1 and FB2, and tumors were harvested 13 days post infection. SAmples were taken from two biological replicates each.

2010-06-20 | E-GEOD-5804 | biostudies-arrayexpress

Role of Ustilago Ncr1 in nitrogen utilization

Project description:The basidiomycete Ustilago maydis is the causal agent of corn smut disease and induces tumor formation during biotrophic growth in its host plant maize. The Usilago maydis genome harbors a homolog to the GATA transcription factors Nit2 and AreA that act as global regulators of nitrogen catabolite repression in filamentous model fungi Neurospora crassa and Aspergillus nidulans. We aimed at resolving the role of the Ustilago maydis homolog Ncr1 for the utilization of complex nitrogen sources and pathogenicity. Sporidia of the indicated Ustilago maydis strains were grown overnight in ammonium minimal medium (Holliday, 1976) and samples for total RNA extraction were taken 2h after transfer to minimal medium lacking any nitrogen source (-N) during the exponential growth phase to assess those genes that are regulated in response to nitrogen starvation. The solopathogenic strain SG200 (control) and deletion mutants of (Nitrogen catabolite repression1) Ncr1 and (Target of Ncr1) Ton1, both being in the SG200 background, were studied in two independent experiments (one experiment for Ton1). Per strain and experiment, three biological replicate samples were analyzed (except for only biological replicates for Ncr1 in the second experiment).

2012-01-01 | E-GEOD-28916 | biostudies-arrayexpress

Ustilago maydis gene expression after growth on hydrophobic surface plus/minus hydroxy fatty acids

Project description:Infection-related development of phytopathogenic fungi is initiated by sensing and responding to plant surface cues. This response results in the formation of specialized infection structures, so-called appressoria. To unravel the program inducing appressoria in the biotrophic smut fungus Ustilago maydis, we exposed cells to a hydrophobic surface and the cutin monomer 16-hydroxy hexadecanoic acid. Genome-wide transcriptional profiling under these appressorium-inducing in vitro conditions revealed dramatic transcriptional changes in almost 20% of the genes. Comparisons with the U. maydis sho1 msb2 double mutant, lacking two putative sensors for plant surface cues, revealed that these plasma membrane receptors regulate a small subset of the surface cue-induced genes. These genes comprised mainly secreted proteins including plant cell wall degrading enzymes that facilitate plant penetration and secreted effectors that are essential virulence factors, with functions after penetration. Our data also demonstrate specific effects on two transcription factors that redirect the transcriptional regulatory network towards appressorium formation and plant penetration. Thus, plant surface cues prime U. maydis for biotrophic development. Solopathogenic Ustilago maydis strain AM1 and its derivate AM1Δsho1Δmsb2 were grown to mid-log phase in YEPSL medium and resuspended in 2% YEPSL plus/minus 16-hydroxy hexadecanoic acid (HDA). The cell suspensions were sprayed on hydrophobic surface (Parafilm) and incubated for 12 h. As control, cells were sprayed on hydrophilic glass surface and incubated for 2 h. After RNA extraction Affymetrix microarrays were performed.

2014-07-23 | E-GEOD-53947 | biostudies-arrayexpress

The WOPR protein Ros1 is a master regulator of sporogenesis and effector gene expression in the maize pathogen Ustilago maydis

Project description:The WOPR protein Ros1 is a master regulator of sporogenesis and effector gene expression in the maize pathogen Ustilago maydis

| PRJNA306676 | ENA

Maize gene expression after infection of Ustilago maydis SG200 and SG200M-NM-^Ttin2

Project description:Anthocyanin induction in plant is considered a general defense response against biotic and abiotic stresses. The infection by Ustilago maydis, the corn smut pathogen, is accompanied with anthocyanin induction in leaf tissue. We revealed that anthocyanin is intentionally induced by the virulence promoting secreted effector protein Tin2. Tin2 protein functions inside plant cells where it interacts with cytoplasmic maize protein kinase ZmTTK1. Tin2 masks an ubiquitin-proteasome degradation motif in ZmTTK1 leading to a more stable active kinase. Active ZmTTK1 controls transcriptional activation of genes in the anthocyanin biosynthesis pathway rerouting phenylalanine away from lignin biosynthesis. Therefore, we performed microarray analysis to understand how maize gene transcription in phenylpropanoid pathway is differentially changed after infection with Ustilago maydis SG200 (wild type) and SG200Dtin2 (anthocyanin-inducing effector mutant). We prepared three biological replicates for mock-inoculated maize (control), SG200-infected maize and SG200M-NM-^Ttin2-infected maize. For 1 sample, we harvested the leaves (1-3cm below injection hole) from 20 plants and pooled them. At 4 days post inoculation, total RNA was extracted.

2014-01-28 | E-GEOD-48536 | biostudies-arrayexpress

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