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

ABSTRACT: 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.

ORGANISM(S): Ustilago maydis

PROVIDER: GSE53947 | GEO | 2014/07/23

SECONDARY ACCESSION(S): PRJNA234013

REPOSITORIES: GEO

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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

Ustilago maydis fox1 mediated gene expression during pathogenic development

2010-12-31 | GSE19591 | GEO

Maize gene expression during infection with Ustilago maydis strain SG200∆fox1

Project description:Ustilago maydis is a plant-pathogenic fungus that establishes a biotrophic relationship with its host Zea mays. The biotrophic interaction is initiated upon host penetration, and involves expansion of the host plasma membrane around hyphae, which is thought to facilitate the exchange of nutrients and virulence factors. Transcriptional regulators involved in the establishment of an infectious dikaryon and penetration into the host have been identified, however, regulators involved in the post-penetration stages remained to be elucidated. In the study we report the identification of an Ustilago maydis forkhead transcription factor, Fox1, which is exclusively expressed during biotrophic development. Deletion of fox1 results in reduced virulence and impaired tumour development in planta. Δfox1 hyphae induce plant defences including the overproduction and accumulation of H2O2 in and around infected cells. This oxidative burst acts as an intercellular signal, which elicits a specific host defence response phenotypically represented by the encasement of proliferating hyphae in extensions of the plant cell wall. Maize microarrays experiments were performed to identify genes involved in the observed plant defence responses on leaf tissue infected with U. maydis strain SG200∆fox1 4 dpi.

2010-12-31 | GSE19559 | GEO

Transcriptome of 3 developmental stages of Colletotrichum graminicola during infection of Zea mays leaf sheaths

Project description:Transcriptome of 3 developmental stages of Colletotrichum graminicola during infection of Zea mays leaf sheaths 3 biological replicates per stage. The three stages are: pre-penetration appressoria (PA), early biotrophic phase (BP), and the switch from biotrophy to necrotrophy (NP). Each biological replicate of the first stage, the pre-penetration appressoria, was sequenced to a 2-fold greater depth due to its lower representation in the samples.

2012-06-04 | E-GEOD-34632 | biostudies-arrayexpress

Ustilago maydis fox1 mediated gene expression during pathogenic development

Project description:Ustilago maydis is a plant-pathogenic fungus that establishes a biotrophic relationship with its host Zea mays. The biotrophic interaction is initiated upon host penetration, and involves expansion of the host plasma membrane around hyphae, which is thought to facilitate the exchange of nutrients and virulence factors. Transcriptional regulators involved in the establishment of an infectious dikaryon and penetration into the host have been identified, however, regulators involved in the post-penetration stages remained to be elucidated. In the study we report the identification of an Ustilago maydis forkhead transcription factor, Fox1, which is exclusively expressed during biotrophic development. Deletion of fox1 results in reduced virulence and impaired tumour development in planta. Microarray analyses of Δfox1-infected plant tissue identified Fox1 as a transcriptional activator, involved in the expression of secreted effectors required for virulence. Maize plants were infected with a mixture of either FB1 and FB2 (wild-type), or FB1∆fox1 and FB2∆fox1 crossings, to measure the impact of fox1 on pathogenic development. Early Golden Bantam maize plants were grown in a phytochamber in a 15h/9h light-dark cycle; light period started/ended with 1h ramping of light intensity. Maize plants were kept at 28°C (light) and 20° (dark). Plantlets were individually sown in pots with potting soil (Fruhstorfer Pikiererde) and infected 7 days after sowing, 1 h before end of the light period. Infected leaf tumor material from at least 10 plants was collected 5 days post infection, 1 h before the end of the light period and directly frozen in liquid nitrogen for RNA-extraction. RNA samples were extracted from infected leaf tissue 5 days after infection.

2010-12-31 | E-GEOD-19591 | biostudies-arrayexpress

Phosphoproteome Analysis of Magnaporthe oryzae During Appressorium Development

Project description:The rice pathogen, Magnaporthe oryzae, undergoes a complex developmental process leading to formation of an appressorium prior to plant infection. In an effort to better understand phosphoregulation during appressorium development, a mass spectrometry-based study was undertaken. Phosphosites were identified from phosphoproteins from mycelia, conidia, germlings, and appressoria of the wild type and a ckpA mutant. The cyclic-AMP dependent protein kinase A (CPKA) is required for initial perception of the leaf surface, development of functional appressoria, and timely mobilization of storage reserves in conidia, including glycogen and lipid bodies.

2015-05-13 | PXD001389 | Pride

Analysis of transcript abundance during appressorium formation in the rice blast fungus Magnaporthe oryzae using HT-SuperSAGE.

Project description:The rice-blast fungus Magnaporthe oryzae is disseminated using a three-celled spore or conidium. Upon landing on the surface of a rice leaf, the conidium germinates and forms a specialised structure for entry into the host plant - the appressorium. In this study we have followed gene expression thoughout germination of the conidium and formation of the appressorium to identify genes that may be important for appressorium function. We have also compared gene expression in the wild-type germinating conidium at four hours to a mutant strain deleted for the MAP kinase pmk1. The pmk1 mutant is unable to form appressoria.

2012-02-13 | GSE30069 | GEO

Magnaporthe appressoria: wt vs. bip1 deletion mutant

Project description:The identification of genes regulated by the transcription factor Bip1 during appressorial development. The rice blast fungus Magnaporthe grisea develops melanized appressoria with high turgor pressure that inject the fungus into epidermal host cells. A screen for M. grisea non-pathogenic mutants recovered mutant M763 with reduced penetration efficiency and infectious growth. Complementation analysis identified a gene inactivated in M763 that encodes a novel basic leucine zipper (bZIP) transcription factor named Bip1 (B-ZIP Involved in Pathogenesis1). Deletion of Bip1 by targeted replacement generated non-pathogenic mutants that are unable to infect rice or barley. These mutants develop melanized appressoria that are unable to penetrate intact or wounded host leaves. Bip1 mRNA was detected in fungal spores, appressoria and infected leaves but not in mycelia, while a BIP1:GFP fusion was detected only in nuclei of mature appressoria. Genome wide transcriptome analysis identified 44 genes down regulated in Δbip1:hph appressoria. Most of these genes (77%) are specifically expressed in appressoria and encode enzymes involved in secondary metabolism (11), secreted proteins and enzymes (18) and G-Protein Coupled Receptors (GPCRs) (5). Promoters of BIP1 down-regulated genes share a similar GCN4/bZIP DNA binding motif (TGACTC). Gel shift assays showed that the motifs from 6 of the 7 promoters tested bind to recombinant BIP1 in vitro, including MGG_08381.5 from the ACE1 locus. Mutation of BIP1 binding sites in the MGG_08381.5 promoter significantly reduced its appressorium specific expression. Molecular and biochemical studies confirmed that BIP1 controls the expression of a distinct set of M. grisea genes in the appressorium, revealing a novel regulatory network involved in early stages of fungal infection that activates a diverse gene expression program resulting in a “shock and awe” impact on the plant host.

2018-01-05 | GSE18823 | GEO

Time series transcriptional profiling of Ustilago maydis infecting Zea mays

Project description:The basidiomycete fungus Ustilago maydis causes smut disease in maize and has become an important model for elucidating the strategies used for host colonization by biotrophic fungi. In this study, we performed an in-depth transcriptional profiling of the plant-associated development of a cross between U. maydis FB1 and FB2 wildtype strains. The analysis of eight different stages, including the development on the leaf surface, early colonization, tumor induction and spore maturation, offers an unprecedented view of the changes in the fungal transcriptome associated with the passage through the entirely biotrophic life cycle. In our analysis, we focus on fungal metabolism, nutritional strategies, secreted effectors and regulatory networks. Secreted proteins were enriched in three distinct expression modules corresponding to the plant surface, establishment of biotrophy and tumor formation, respectively. These modules are likely the key determinants for U. maydis virulence. With respect to nutrient utilization, we observed that expression of several nutrient transporters was tied to these virulence modules rather than being controlled by nutrient availability. We show that oligopeptide transporters likely involved in nitrogen supply during infection are important virulence determinants. By measuring the intramodular connectivity of transcription factors, we identified potential drivers for the virulence modules. While known components of the b-cascade served as inducers for the plant surface and biotrophy module, we identified a set of yet uncharacterized transcription factors as likely responsible for expression of the tumor module. We demonstrate a crucial role in effector gene expression and tumor formation for one of these transcription factors.

2018-01-11 | GSE103876 | GEO

Analysis of transcript abundance during appressorium formation in the rice blast fungus Magnaporthe oryzae using HT-SuperSAGE.

2012-02-13 | E-GEOD-30069 | biostudies-arrayexpress

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