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Leucine metabolism regulates TRI6 expression and affects deoxynivalenol production and virulence in Fusarium graminearum

ABSTRACT: TRI6 is a positive regulator of the trichothecene gene cluster and the production of trichothecene mycotoxins (deoxynivalenol [DON] and acetylated forms such as 15-ADON) in the cereal pathogen F. graminearum. As a global transcriptional regulator, TRI6 expression is modulated by nitrogen-limiting conditions, sources of nitrogen and carbon, pH, and light. However, the mechanism by which these diverse environmental factors affect TRI6 expression remains under-explored. In our effort to understand how nutrients affect TRI6 regulation, comparative digital expression profiling was performed with a wildtype F. graminearum and a Δtri6 mutant strain, grown in nutrient-rich conditions. Analysis showed that TRI6 negatively regulates genes of the branched-chain amino acid (BCAA) metabolic pathway. Feeding studies with deletion mutants of MCC, encoding methylcrotonyl-CoA-carboxylase, one of the key enzymes of leucine metabolism, showed that addition of leucine specifically down regulated TRI6 expression and reduced 15-ADON accumulation. Constitutive expression of TRI6 in the Δmcc mutant strain restored 15-ADON production. A combination of cellophane breach assays and pathogenicity experiments on wheat demonstrated that disrupting the leucine metabolic pathway significantly reduced disease. These findings suggest a complex interaction between one of the primary metabolic pathways with a global regulator of mycotoxin biosynthesis and virulence in F. graminearum.

ORGANISM(S): Fusarium graminearum

PROVIDER: GSE71604 | GEO | 2015/08/01

SECONDARY ACCESSION(S): PRJNA291605

REPOSITORIES: GEO

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Leucine metabolism regulates TRI6 expression and affects deoxynivalenol production and virulence in Fusarium graminearum

Project description:TRI6 is a positive regulator of the trichothecene gene cluster and the production of trichothecene mycotoxins (deoxynivalenol [DON] and acetylated forms such as 15-ADON) in the cereal pathogen F. graminearum. As a global transcriptional regulator, TRI6 expression is modulated by nitrogen-limiting conditions, sources of nitrogen and carbon, pH, and light. However, the mechanism by which these diverse environmental factors affect TRI6 expression remains under-explored. In our effort to understand how nutrients affect TRI6 regulation, comparative digital expression profiling was performed with a wildtype F. graminearum and a Δtri6 mutant strain, grown in nutrient-rich conditions. Analysis showed that TRI6 negatively regulates genes of the branched-chain amino acid (BCAA) metabolic pathway. Feeding studies with deletion mutants of MCC, encoding methylcrotonyl-CoA-carboxylase, one of the key enzymes of leucine metabolism, showed that addition of leucine specifically down regulated TRI6 expression and reduced 15-ADON accumulation. Constitutive expression of TRI6 in the Δmcc mutant strain restored 15-ADON production. A combination of cellophane breach assays and pathogenicity experiments on wheat demonstrated that disrupting the leucine metabolic pathway significantly reduced disease. These findings suggest a complex interaction between one of the primary metabolic pathways with a global regulator of mycotoxin biosynthesis and virulence in F. graminearum. Triplicate samples of digital gene expression profiling data for the Tri6 mutant and wildtype strains were compared. **Please note that the raw data files for the current study are no longer available, and therefore are not included in the submission.

2015-08-01 | E-GEOD-71604 | biostudies-arrayexpress

Tri6 is a global transcription regulator in the phytopathogen Fusarium graminearum

Project description:In F. graminearum, the transcriptional regulator TRI6 is encoded within the trichothecene gene cluster and regulates genes involved in the biosynthesis of the secondary metabolite deoxynivalenol (DON). Targeted disruption of TRI6 confirmed its role as a positive regulator of trichothecene genes and previous studies designated Tri6 as a pathway-specific transcriptional regulator. The Tri6 protein with its Cys2His2 zinc-finger may also conform to the class of broad-domain transcription regulators. This class of global transcriptional regulators mediate various environmental cues and generally responds to the demands of cellular metabolism. Expression profiling of F. graminearum grown under nitrogen-limiting conditions revealed that 49 out of 198 target genes are differentially regulated by TRI6. The identification of potential new targets together with deciphering novel binding site for Tri6, casts new light into the role of this transcriptional regulator in the overall growth and development of F. graminearum. Three biological replicates of Fusarium graminearum wildtype strain GZ3639 (NRRL 38155) (reference) and a tri6∆ mutant derived from GZ3639 were grown under nitrogen-limiting conditions in liquid culture for 5 hrs at 28oC

2011-09-30 | E-GEOD-30892 | biostudies-arrayexpress

Tri6 is a global transcription regulator in the phytopathogen Fusarium graminearum

2011-10-01 | GSE30892 | GEO

Gene expression in wheat heads treated with the mutant tri6Δ of Fusarium graminearum, which produces a priming-induced resistance to fusarium head blight

2015-06-02 | GSE69437 | GEO

Gene expression in wheat heads treated with the mutant noxABΔ of Fusarium graminearum, which produces a priming-induced resistance to fusarium head blight

Project description:Fusarium head blight (FHB) is a major disease of cereal crops caused by the fungus Fusarium graminearum (Fg). FHB affects the flowering heads (or spikes) and developing seeds. This study compares the gene expression profile of heads from the susceptible cultivar Roblin after inoculation with either water (dH2O) or the disarmed Fg strain noxABΔ (NoxAB). noxABΔ is a double deletion mutant of NoxA and NoxB, which function are component of the NADPH oxidase enzyme complex converting molecular oxygen in a stepwise reduction to superoxides leading to the production of H2O2. Deletion mutants of Nos A and B have been described in more details in Li Wang, Christopher Mogg, Sean Walkowiak, Manisha Joshi & Rajagopal Subramaniam (2014). Characterization of NADPH oxidase genes NoxA and NoxB in Fusarium graminearum. Can. J. Plant Pathol. 36: 12-21, DOI: 10.1080/07060661.2013.868370. This manuscript characterizes the role of the catalytic subunit (gp91phox) of Nox, NoxA and NoxB in F. graminearum. Targeted deletion of NoxA and NoxB show that these genes differentially regulate the production of superoxides during mycelia development. The nitro blue tetrazolium staining revealed that both the single noxA mutant and the double noxA/B mutant strains are restricted in the production of superoxides. This limitation, however, did not affect their ability to synthesize 15-ADON in culture. Deletion analyses also revealed that NoxA, but not NoxB is involved in perithecia development and ascospore production. An in-vitro based cellophane breach assay indicated that both NoxA and NoxB contribute to virulence. The pathogenicity tests performed on a susceptible variety of wheat, ‘Roblin’ confirmed that these two genes act synergistically to promote virulence. Cumulatively, we provide evidence that the Nox homologues have non-redundant functions in F. graminearum.

2015-06-02 | GSE69490 | GEO

Differential transcriptome analyses of three wheat genotypes in their response to Fusarium Head Blight and trichothecenes

Project description:Fusarium Head Blight (FHB) is a disease of wheat and other cereal crops, where Fusarium graminearum and related species infects the wheat inflorescence during and post-anthesis. The fungus produces trichothecene toxins that accumulate in the grain of infected head, and are required for disease spread. Microarrays were used to observe differential gene expression in the uninoculated spikelets of FHB-challenged wheat spikes in three wheat genotypes. A summary of our findings will be published in Plant Pathology. Three wheat genotypes were used: (1) 'Superb', an FHB-susceptible Canadian wheat cultivar; (2) GS-1-EM0040 (CIMMYT11x'Superb'*2), a double haploid line with good resistance to initial infection (Type 1 resistance), and moderate resistance to disease spread (Type 2 resistance); and (3) GS-1-EM0168 (CM82036x'Superb'*2), a double haploid line with moderate Type 1 resistance, and good Type 2 resistance. Five inocula were used: (A) water, (B) FgTri5+ (GZ3639, a trichothecene-producing F. graminearum strain); (C) FgTri5- (GZT40, a trichothecene-non-producing mutant of the F. graminearum strain GZ3639); (D) FgTri5 supplemented with deoxynivalenol (DON), which is the main trichothecene produced by FgTri5+; and (E) DON. The inocula were injected into two spikelets near the center of the spike during early stages of anthesis, and spikelets above and below the inoculation point were collected at 3, 8, and 24 h after inoculation. A zero-hour un-inoculated control was also collected from each line. Total RNA was extracted from collected spikelets, and microarray analysis was perfomed using the Affymetrix Wheat GeneChip.

2011-08-31 | E-GEOD-28973 | biostudies-arrayexpress

The Transcription factor FgSfp1 orchestrates mycotoxin deoxynivalenol biosynthesis in Fusarium graminearum

Project description:Fusarium graminearum (F. graminearum) and its derivative mycotoxin deoxynivalenol (DON) are highly concerned with food security, safety and sustainability worldwide. The molecular mechanism regulates DON biosynthesis in F. graminearum remains enigmatic, despite several transcription factors (TFs) have been revealed containing regulatory functions. Here, we first characterized a zinc finger-contained TF, FgSfp1. Interestingly, contrast to the previous knowledge, all TRI-cluster genes were abnormally upregulated in ∆FgSfp1 while Tri proteins abundance rationally decreased, resulting in a 95.4% reduction of DON yield simultaneously. Further evidence determined FgSfp1 acted as a nutrient-sensing factor coordinates genetic expression efficiency through interference of ribosomal biogenesis process. Specifically, FgSfp1-depletion leads to ribosome biogenesis assembly factor (RiBi) expression attenuation along with DON precursor acetyl-CoA synthase reduction since FgSfp1 actively interacts with RNA 2’-O-methylation enzyme FgNop1 revealed by Bi-FC and subsequently influences mRNA translation capacity. In conclusion, we elucidated that the FgSfp1 orchestrates DON biosynthesis via participating RNA posttranscriptional modification for ribosomal RNA maturation, offering new insights into the DON biosynthesis regulation. Ultimately, this novel TF might be a key regulator for DON contamination control in the whole food chain.

2024-11-06 | GSE262607 | GEO

GC-MS based targeted metabolic profiling identifies changes in the wheat metabolome following deoxynivalenol treatment

Project description:Fusarium graminearum and related species commonly infest grains causing the devastating plant disease Fusarium head blight (FHB) and the formation of trichothecene mycotoxins. The most relevant toxin is deoxynivalenol (DON), which acts as a virulence factor of the pathogen. FHB is difficult to control and resistance to this disease is a polygenic trait, mainly mediated by the quantitative trait loci (QTL) Fhb1 and Qfhs.ifa-5A. In this study we established a targeted GC-MS based metabolomics workflow comprising a standardized experimental setup for growth, treatment and sampling of wheat ears and subsequent GC-MS analysis followed by data processing and evaluation of QC measures using tailored statistical and bioinformatics tools. This workflow was applied to wheat samples of six genotypes with varying levels of Fusarium resistance, treated with either DON or water, and harvested 0, 12, 24, 48 and 96 hours after treatment. The results suggest that the primary carbohydrate metabolism and transport, the citric acid cycle and the primary nitrogen metabolism of wheat are clearly affected by DON treatment. Most importantly significantly elevated levels of amino acids and derived amines were observed. In particular, the concentrations of the three aromatic amino acids phenylalanine, tyrosine, and tryptophan increased. No clear QTL specific difference in the response could be observed except a generally faster increase in shikimate pathway intermediates in genotypes containing Fhb1. The overall workflow proved to be feasible and facilitated to obtain a more comprehensive picture on the effect of DON on the central metabolism of wheat.

2014-10-17 | MTBLS112 | MetaboLights

Transcriptome analysis of a wheat NIL pair carrying FHB resistant and susceptible alleles

Project description:In this study, we used the Affymetrix wheat GeneChip to examine the transcript accumulation in a near-isogenic line pair carrying resistant and susceptible alleles at the wheat Fhb1 locus. The objectives of this study were: (1) to identify the overall response in wheat to F. graminearum infection; (2) to identify key genes involved in FHB resistance/susceptibility pathways in wheat; (3) to compare the transcript profiles of wheat and barley during F. graminearum infection; and (4) to examine the relationship between transcript accumulation, disease severity, fungal biomass and trichothecene accumulation in wheat. ****[PLEXdb(http://www.plexdb.org) has submitted this series at GEO on behalf of the original contributor, Seungho Cho. The equivalent experiment is TA20 at PLEXdb.] FHB resistant and susceptible plants 48 an 96 hours after treatment with Fusarium graminearum strain Butte 86

2011-08-30 | E-GEOD-31758 | biostudies-arrayexpress

Dual RNA-sequencing of Fusarium Head Blight resistance in winter wheat

Project description:In this study, we used dual RNA-sequencing to profile FHB-resistant AC Emerson, FHB-moderately AC Morley, and FHB-susceptible CDC Falcon winter wheat cultivars prior to and in response to Fusarium graminearum at 7 days post inoculation. Differential expression analyses revealed distinct defense responses between resistant and susceptible wheat cultivars including increased mechanical defense through lignin biosynthesis and increased deoxynivalenol (DON) detoxification through UDP-glycosyltransferase activity in resistant cultivars. Further, differential expression analysis in F. graminearum challenging these distinct cultivars revealed changes genes involved in trichothecene mycotoxin biosynthesis.

2024-01-24 | GSE233409 | GEO

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