Gene expression during biotrophic growth
Ontology highlight
ABSTRACT:
ORGANISM(S): Ustilago maydis
SUBMITTER: Mario Scherer
PROVIDER: E-GEOD-5804 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
Similar Datasets
Project description:Identification of transcriptional changes of Ustilago maydis wild type strain FB1 grown in axenic culture in complete medium containing glucose without or with the addition of 10 µM FeSO4. Keywords: gene induction Strain FB1 (wt) was grown in CM glucose medium in the presence or absence of 10 µM FeSO4.
2010-06-12 | E-GEOD-6037 | biostudies-arrayexpress
Project description:Urbs1 of Ustilago maydis is a transcription factor that has been shown to repress its target genes (e.g. sid1, sid2) in the presence of iron. To identify additional genes regulated by Urbs1, we compared transcriptional profiles of the Ustilago maydis wild type strain FB1 grown in complete medium containing glucose with the addition of 10 µM FeSO4 to the Ustilago maydis urbs1 deletion mutant BW12 grown under the same conditions. Keywords: gene induction Strains FB1 (control) and BW12 (urbs1 deletion mutant) were grown in CM glucose in the presence of iron.
2010-06-12 | E-GEOD-6038 | biostudies-arrayexpress
Project description:In fungi, sexual compatibility is controlled by mating type loci that prevent self-fertilization. In the plant pathogenic fungus Ustilago maydis, the b mating type locus encodes a pair of unrelated homeodomain proteins, termed bE and bW. After fusion of two compatible, haploid cells, the bE and bW proteins form a heterodimeric complex, but only if they are derived from different, compatible alleles. The active bE/bW complex is required and sufficient to initiate pathogenic development of U. maydis, which is prerequisite for sexual reproduction of the fungus. However, the role of the b heterodimer during later stages of pathogenic development was unclear. To analyze b function during in planta development, we generated a temperature-sensitive bE allele (bEts) encoding a protein with a single amino acid alteration at the border of the homeodomain. This mutation leads to a stop in pathogenic development at the restrictive temperature in planta, while bEts strains show normal development at permissive temperature. At restrictive temperature, hyphae develop enlarged, bulbous cells at their tips that contain multiple nuclei, indicating a severe defect in cell division. DNA array analysis of bEts mutant strains in planta revealed a b-dependent regulation of genes coding for secreted proteins that were shown to influence fungal virulence. Our data demonstrate that in U. maydis the b heterodimer is not only essential to establish the heterodikaryon after mating of two compatible sporidia and to initiate fungal pathogenicity, but also to sustain in planta proliferation and ensure sexual reproduction. Maize plants were infected with a mixture of either FB1 and FB2 (wild-type), or RAb1ts and RAb2ts (temperature-sensitive b heterodimer) and kept at 22°C (permissive conditions). Control samples of FB1/FB2 and RAb1ts and RAb2ts were taken 5 days post inoculation at 22 °C (1 replicate each), to see if strains expressing the bts heterodimer show wildtype-like biotrophic development. To measure the impact of the b heterodimer on pathogenic development, infected plants were shifted for 9 hours to resrictive conditions (31°C) 111 hours post infection. After the temperature shift FB1/FB2 (3 replicates) infections developed normal, whereas RAb1ts/RAb2ts (3 replicates) infections were not able to further proliferate in planta because of a none-functional b heterodimer.
2010-05-18 | E-GEOD-16501 | biostudies-arrayexpress
Project description:In mammalian cells RACK1 serves as a scaffold protein that has a role in integrating inputs from different signaling pathways and affects translation through association with ribosomes. U. maydis contains a seven-WD40 repeat motif protein designated Rak1, which shows 68% identity to RACK1 and is homologous to Asc1p of Saccharomyces cerevisiae. The asc1 mutant could be complemented by introduction of U. maydis rak1. The deletion of rak1 affected cell growth, cell wall integrity and specifically attenuated cell fusion. This latter defect was caused by reduced expression of prf1 encoding the regulator for pheromone and pheromone-receptor genes. Rak1 interacts with a variety of ribosomal proteins and microarray analysis revealed that the deletion of rak1 led to severely reduced expression of rop1, an activator prf1. The constitutive expression of rop1 could rescue the defect of mfa1 expression as well as conjugation tube formation in response to pheromone induction in the rak1 mutant. Moreover, a solopathogenic rak1 mutant failed to respond to plant derived stimuli, resulting in attenuated filamentation and pathogenicity. This could be partially rescued by constitutive expression of the b heterodimer. These data suggest that rak1 is a regulator of rop1 expression with additional roles after cell fusion. Ustilago maydis strains FB1 and FB1?rak1 were grown on a rotary shaker (200 rpm) in liquid Ustilago complete medium with 1% glucose at 28°C to an OD600 of 0.5. The experiment was performed using three independent biological replicates per strain.
2011-07-05 | E-GEOD-30382 | biostudies-arrayexpress
Project description:Adr1 of Ustilago maydis is a protein kinase that is activated after separation from the regulatory subunit mediated by high cAMP levels. A copy of the gene under the control of the arabinose-inducible crg1 promotor was introduced into the Cbx locus of the wild type strain FB1 creating strain HE140. To monitor the transcriptional changes upon adr1 induction by arabinose, we conducted a time course experiment comparing the wild type transcriptional response before and 75 or 180 min after medium shift to complete medium containing arabinose, as well as the response of the mutant HE140 before and 75 or 180 min after induction of the additional copy of adr1 by arabinose. Keywords: gene induction Strains FB1 (control) and HE140 (a1b1 ipr[pcrg1:adr1]ips) were grown in CM medium containing glucose and were shifted to CM medium containing arabinose and grown for 75 or 180 min. Samples were taken before and after arabinose induction from two (glucose) or three (arabinose) biological replicates each.
2010-06-12 | E-GEOD-6039 | biostudies-arrayexpress
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
Project description:Identification of transcriptional changes of Ustilago maydis wild type strain FB1 grown in axenic culture in complete medium containing glucose without or with the addition of 10 µM FeSO4. Keywords: gene induction
2007-01-01 | GSE6037 | GEO
Project description:This SuperSeries is composed of the following subset Series: GSE18750: Controlled expression of compatible and incompatible combinations of Ustilago maydis b-mating type locus genes bE and bW GSE18754: Effect of rbf1 deletion during controlled expression of of Ustilago maydis b-mating type locus genes bE1 and bW2 GSE18756: Rbf1 induced gene expression in Ustilago maydis Refer to individual Series
2010-06-01 | E-GEOD-18881 | biostudies-arrayexpress
Project description:We compared multiple strains of lab trophozoites to recent clinical isolates. Clinical isolates were grown in xenic media, and maintained many characteristics of the cyst stage of devlopment Keywords: Stage conversion E. histolytica trophozoites of three different strains (HM1:IMSS, Rahman, and 200:NIH) and grown under multiple conditions (Long term axenic growth in TYI, Growth in low glucose media, Mouse adapted parasites and in vivo growth in a mouse model) were compared to recent clinical isolates that had grown in xenic culture for 1-8 weeks.
2007-03-01 | E-GEOD-6648 | biostudies-arrayexpress
Project description:The fungus Ustilago maydis is a biotrophic pathogen of corn. In its genome we have identified an ortholog of YAP1 from Saccharomyces cerevisae which regulates the oxidative stress response in this organism. yap1 mutants of U. maydis displayed higher sensitivity to H2O2 than wild type cells and their virulence was significantly reduced. U. maydis yap1 could partially complement the H2O2 sensitivity of a yap1 deletion mutant of S. cerevisiae and a Yap1-GFP fusion protein showed nuclear localization after H2O2 treatment, suggesting that Yap1 in U. maydis functions as a redox sensor. Mutations in two cysteine residues prevented accumulation in the nucleus and the respective mutant strains showed the same virulence phenotype as Dyap1 mutants. DAB staining revealed an accumulation of H2O2 around yap1 mutant hyphae which was absent in wild type. Inhibition of the plant NADPH oxidase prevented this accumulation and restored virulence. During the infection Yap1 showed nuclear localization after penetration up to 2-3 days after infection. Through array analysis a large set of yap1 regulated genes were identified and these included two peroxidase genes. Deletion mutants of these genes were attenuated in virulence. These results suggest that U. maydis is using its yap1 controlled H2O2 detoxification system for coping with early plant defense responses. Keywords: Oxidative stress, yap1 dependent genes, Ustilago maydis Two independent overnight cultures of U. mayidis FB1 and FB1Dyap1 grown in CM-glucose (OD600 0.8) were diluted in 100 ml of the same medium (OD600 0.2) and growth at 28° C until an OD600 0.6. The cultures were divided and one half was supplemented with 5mM H2O2. After one hour of exposition to H2O2, cells were harvested by centrifugation and frozen in liquid nitrogen. RNA extraction, purification, cDNA generation, purification and labeling were performed according to standard protocols (Affymetrix). DNA array analysis was performed performed on two biological replicates each, using custom-designed Affymetrix chips (MPIUstilagoA). Data were analysed using a GeneArray Scanner (Agilent/Affymetrix) and the GeneChip Expression Analysis software (GCOS)
2010-06-11 | E-GEOD-7518 | biostudies-arrayexpress