Isolation, characterization and disruption of the areA nitrogen regulatory gene of Gibberella fujikuroi

Tudzynski, Bettina; Homann, Veronika; Feng, Bo; Marzluf, George A.

doi:10.1007/s004380050947

Cited by 133 publications

(77 citation statements)

References 48 publications

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“…Generation of protoplasts and transformation of F. fujikuroi were carried out according to (Tudzynski et al, 1999). Regeneration of transformed protoplasts was performed for 4–5 days at 28°C in a regeneration medium (0.7 M sucrose, 0.05% yeast extract) containing either 100 mg/mL nourseothricin (Werner-Bioagents, Jena, Germany) or 100 mg/mL hygromycin (Calbiochem, Darmstadt, Germany).…”

Section: Methodsmentioning

confidence: 99%

The GATA-Type Transcription Factor Csm1 Regulates Conidiation and Secondary Metabolism in Fusarium fujikuroi

et al. 2017

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GATA-type transcription factors (TFs) such as the nitrogen regulators AreA and AreB, or the light-responsive TFs WC-1 and WC-2, play global roles in fungal growth and development. The conserved GATA TF NsdD is known as an activator of sexual development and key repressor of conidiation in Aspergillus nidulans, and as light-regulated repressor of macroconidia formation in Botrytis cinerea. In the present study, we functionally characterized the NsdD ortholog in Fusarium fujikuroi, named Csm1. Deletion of this gene resulted in elevated microconidia formation in the wild-type (WT) and restoration of conidiation in the non-sporulating velvet mutant Δvel1 demonstrating that Csm1 also plays a role as repressor of conidiation in F. fujikuroi. Furthermore, biosynthesis of the PKS-derived red pigments, bikaverin and fusarubins, is de-regulated under otherwise repressing conditions. Cross-species complementation of the Δcsm1 mutant with the B. cinerea ortholog LTF1 led to full restoration of WT-like growth, conidiation and pigment formation. In contrast, the F. fujikuroi CSM1 rescued only the defects in growth, the tolerance to H2O2 and virulence, but did not restore the light-dependent differentiation when expressed in the B. cinerea Δltf1 mutant. Microarray analysis comparing the expression profiles of the F. fujikuroi WT and the Δcsm1 mutant under different nitrogen conditions revealed a strong impact of this GATA TF on 19 of the 47 gene clusters in the genome of F. fujikuroi. One of the up-regulated silent gene clusters is the one containing the sesquiterpene cyclase-encoding key gene STC1. Heterologous expression of STC1 in Escherichia coli enabled us to identify the product as the volatile bioactive compound (–)-germacrene D.

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Section: Methodsmentioning

confidence: 99%

The GATA-Type Transcription Factor Csm1 Regulates Conidiation and Secondary Metabolism in Fusarium fujikuroi

et al. 2017

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“…The F. fujikuroi Wt, the newly generated knockout strains and the already available Δ NIAD (Tudzynski et al, 1996), Δ AREA (Tudzynski et al, 1999), and Δ AREB (Michielse et al, 2014) deletion mutants were incubated for 4 days on solidified synthetic ICI minimal medium, supplemented with different nitrogen sources ( Figure 1 ). All mutant strains were able to grow in a Wt-like manner on glutamine, with the exception of Δ AREB , which showed a slightly reduced growth.…”

Section: Resultsmentioning

confidence: 99%

“…In this study the following F. fujikuroi strains were used: Wt strain IMI58289 (Commonwealth Mycological Institute, Kew, UK), Δ AREA -T19 (Tudzynski et al, 1999) and Δ AREB -T2.1 (Michielse et al, 2014). Strains were maintained on solid CM (Pontecorvo et al, 1953) and cultivated at 28°C in darkness.…”

Section: Methodsmentioning

confidence: 99%

Nitrate Assimilation in Fusarium fujikuroi Is Controlled by Multiple Levels of Regulation

2017

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Secondary metabolite production of the phytopathogenic ascomycete fungus Fusarium fujikuroi is greatly influenced by the availability of nitrogen. While favored nitrogen sources such as glutamine and ammonium are used preferentially, the uptake and utilization of nitrate is subject to a regulatory mechanism called nitrogen metabolite repression (NMR). In Aspergillus nidulans, the transcriptional control of the nitrate assimilatory system is carried out by the synergistic action of the nitrate-specific transcription factor NirA and the major nitrogen-responsive regulator AreA. In this study, we identified the main components of the nitrate assimilation system in F. fujikuroi and studied the role of each of them regarding the regulation of the remaining components. We analyzed mutants with deletions of the nitrate-specific activator NirA, the nitrate reductase (NR), the nitrite reductase (NiR) and the nitrate transporter NrtA. We show that NirA controls the transcription of the nitrate assimilatory genes NIAD, NIIA, and NRTA in the presence of nitrate, and that the global nitrogen regulator AreA is obligatory for expression of most, but not all NirA target genes (NIAD). By transforming a NirA-GFP fusion construct into the ΔNIAD, ΔNRTA, and ΔAREA mutant backgrounds we revealed that NirA was dispersed in the cytosol when grown in the presence of glutamine, but rapidly sorted to the nucleus when nitrate was added. Interestingly, the rapid and nitrate-induced nuclear translocation of NirA was observed also in the ΔAREA and ΔNRTA mutants, but not in ΔNIAD, suggesting that the fungus is able to directly sense nitrate in an AreA- and NrtA-independent, but NR-dependent manner.

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“…Therefore, knock-out fragments were amplified with the following primer pairs: FgCCL1_5F and genR_splitF as well as genR_splitR and FgCCL1_3R. All transformed protoplasts were regenerated as described (Tudzynski et al, 1999). The medium contained 100 ppm of the appropriate resistance marker.…”

Section: Methodsmentioning

confidence: 99%

Lack of the COMPASS Component Ccl1 Reduces H3K4 Trimethylation Levels and Affects Transcription of Secondary Metabolite Genes in Two Plant–Pathogenic Fusarium Species

Studt

Janevska²,

Arndt

et al. 2017

Front. Microbiol.

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In the two fungal pathogens Fusarium fujikuroi and Fusarium graminearum, secondary metabolites (SMs) are fitness and virulence factors and there is compelling evidence that the coordination of SM gene expression is under epigenetic control. Here, we characterized Ccl1, a subunit of the COMPASS complex responsible for methylating lysine 4 of histone H3 (H3K4me). We show that Ccl1 is not essential for viability but a regulator of genome-wide trimethylation of H3K4 (H3K4me3). Although, recent work in Fusarium and Aspergillus spp. detected only sporadic H3K4 methylation at the majority of the SM gene clusters, we show here that SM profiles in CCL1 deletion mutants are strongly deviating from the wild type. Cross-complementation experiments indicate high functional conservation of Ccl1 as phenotypes of the respective △ccl1 were rescued in both fungi. Strikingly, biosynthesis of the species-specific virulence factors gibberellic acid and deoxynivalenol produced by F. fujikuroi and F. graminearum, respectively, was reduced in axenic cultures but virulence was not attenuated in these mutants, a phenotype which goes in line with restored virulence factor production levels in planta. This suggests that yet unknown plant-derived signals are able to compensate for Ccl1 function during pathogenesis.

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Isolation, characterization and disruption of the areA nitrogen regulatory gene of Gibberella fujikuroi

Cited by 133 publications

References 48 publications

The GATA-Type Transcription Factor Csm1 Regulates Conidiation and Secondary Metabolism in Fusarium fujikuroi

The GATA-Type Transcription Factor Csm1 Regulates Conidiation and Secondary Metabolism in Fusarium fujikuroi

Nitrate Assimilation in Fusarium fujikuroi Is Controlled by Multiple Levels of Regulation

Lack of the COMPASS Component Ccl1 Reduces H3K4 Trimethylation Levels and Affects Transcription of Secondary Metabolite Genes in Two Plant–Pathogenic Fusarium Species

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