Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Role of Ustilago Ncr1 in nitrogen utilization

ABSTRACT: 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).

ORGANISM(S): Ustilago maydis

SUBMITTER: Lars Voll

PROVIDER: E-GEOD-28916 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Many of the genes coding for secreted protein effectors are arranged in gene clusters in the genome of the biotrophic plant pathogen Ustilago maydis. The largest of these gene clusters, cluster 19A, encodes 24 secreted effectors. Deletion of the entire cluster results in severe attenuation of virulence. The generation and analysis strains carrying sub-deletions identified 9 genes significantly contributing to tumor formation after seedling infection. As the individual contributions of these genes to tumor formation were small, we studied the response of maize plants to the whole cluster mutant as well as to several individual mutants by array analysis. This revealed distinct plant responses, demonstrating that the respective effectors have discrete plant targets. Many of the genes coding for secreted protein effectors are arranged in gene clusters in the genome of the biotrophic plant pathogen Ustilago maydis. The largest of these gene clusters, cluster 19A, encodes 24 secreted effectors. Deletion of the entire cluster results in severe attenuation of virulence. The generation and analysis strains carrying sub-deletions identified 9 genes significantly contributing to tumor formation after seedling infection. As the individual contributions of these genes to tumor formation were small, we studied the response of maize plants to the whole cluster mutant as well as to several individual mutants by array analysis. This revealed distinct plant responses, demonstrating that the respective effectors have discrete plant targets. We used the Affymetrix maize genome array to analyze the transcriptional responses of maize to cluster 19A mutants and individual sub-deletions for the cluster 19A genes tin1, tin3, tin4 and tin5. We found plant responses to the mutants were significantly different although the macroscopic phenotypes of the individual mutants were very similar. U. maydis infected parts of maize seedling leaves were dissected 4 days after inoculation with strain SG200∆19A, SG200∆tin1, SG200∆tin3, SG200∆tin4 and SG200∆tin5, respectively. We previously submitted data of maize leaves that were treated with the progenitor wild type strain SG200 as well as mock-infections under identical experimetal conditions (GEO: GSE10023). These data served as controls for this experiment.

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Role of Ustilago Ncr1 in nitrogen utilization

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