<i>Fusarium graminearum</i> Tri12p Influences Virulence to Wheat and Trichothecene Accumulation

Menke, Jon; Dong, Ya Nhong; Kistler, H. Corby

doi:10.1094/mpmi-04-12-0081-r

Cited by 88 publications

(112 citation statements)

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“…In addition, microscopic analysis showed that the g8 mutant exhibited strong TRI5-GFP signals in thickened coralloid hyphae under repressive conditions similar to those only observed under inductive conditions in the TRI5-GFP parental strain. These toxin-production-specific structures, as described by Menke et al (2012), developed in the mutant under repressive conditions, but their complete differentiation was impaired, even under inductive conditions.…”

Section: Discussionmentioning

confidence: 92%

“…Previous studies by Menke et al (2012) have shown that F. graminearum develops specific subapical ovoid cell structures and highly branched thick coralloid hyphae under toxin production inductive conditions in vitro. These cellular structures are similar to infection-related cellular structures observed in in planta studies (Pritsch et al, 2000;Jansen et al, 2005;Boddu et al, 2006;Ilgen et al, 2009;Rittenour and Harris, 2010;Kistler and Broz, 2015).…”

Section: The G8 Mutant Develops Toxin-production-specific Cellular Stmentioning

confidence: 94%

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High-throughput FACS-based mutant screen identifies a gain-of-function allele of the Fusarium graminearum adenylyl cyclase causing deoxynivalenol over-production

Blum

Benfield

Stiller

et al. 2016

Fungal Genetics and Biology

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., High-throughput FACS-based mutant screen identifies a gain-of-function allele of the Fusarium graminearum adenylyl cyclase causing deoxynivalenol over-production, Fungal Genetics and Biology (2016), doi: http://dx.doi. org/10.1016/j.fgb. 2016.02.005 This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. ABSTRACTFusarium head blight and crown rot, caused by the fungal plant pathogen Fusarium graminearum, impose a major threat to global wheat production. During the infection, plants are contaminated with mycotoxins such as deoxynivalenol (DON), which can be toxic for humans and animals. In addition, DON is a major virulence factor during wheat infection. However, it is not fully understood how DON production is regulated in F. graminearum. In order to identify regulators of DON production, a high-throughput mutant screen using Fluorescence Activated Cell Sorting (FACS) of a mutagenized TRI5-GFP reporter strain was established and a mutant over-producing DON under repressive conditions identified. A gain-of-function mutation in the F. graminearum adenylyl cyclase (FAC1), which is a known positive regulator of DON production, was identified as the cause of this phenotype through genome sequencing and segregation analysis. Our results show that the high-throughput mutant screening procedure developed here can be applied for identification of fungal proteins involved in diverse processes.

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Section: Discussionmentioning

confidence: 92%

Section: The G8 Mutant Develops Toxin-production-specific Cellular Stmentioning

confidence: 94%

High-throughput FACS-based mutant screen identifies a gain-of-function allele of the Fusarium graminearum adenylyl cyclase causing deoxynivalenol over-production

Blum

Benfield

Stiller

et al. 2016

Fungal Genetics and Biology

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“…As already stated by Menke et al (2012) as well as in the first chapter here, the morphological changes of hyphae observed during toxin production in vitro are similar to those observed during plant infection (Boddu et al, 2006;Brown et al, 2010;Ilgen et al, 2009;Jansen et al, 2005;Pritsch et al, 2000;Rittenour and Harris, 2010). The bulbous hyphae resemble infection hyphae which can be found growing within the infected plant tissue, however the actual function of the ovoid cells observed in vitro is still unknown.…”

Section: Cellular Developmental Changes and Subcellular Reorganizatiosupporting

confidence: 84%

“…The g8 mutant develops toxin-production-specific cellular structures under toxin production repressive conditions Previous studies by Menke et al (2012) have shown that F. graminearum develops specific subapical ovoid cell structures and highly branched thick coralloid hyphae under toxin production inductive conditions in vitro. These cellular structures are similar to infection-related cellular structures observed in in planta studies (Boddu et al, 2006;Ilgen et al, 2009;Jansen et al, Kistler and Broz, 2015;Pritsch et al, 2000;Rittenour and Harris, 2010;).…”

Section: 34mentioning

confidence: 99%

“…A further member of the TRI5 gene cluster is TRI12, which encodes a potential trichothecene efflux pump with 14 predicted transmembrane spanning regions (Alexander et al, 1999;Wuchiyama et al, 2000). TRI12 is required for auto-tolerance of F. graminearum to growth inhibition under toxin production inducing conditions (Menke et al, 2012).…”

Section: Don Biosynthesis In F Graminearummentioning

confidence: 99%

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Regulation and cell biology of secondary metabolite production in Fusarium graminearum and Fusarium pseudograminearum

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1 AbstractAscomycete fungi have the potential to produce a vast array of secondary metabolites, which are metabolites not required for normal growth and development. In any one species there is typically the potential for production of upward of fifty of these compounds. Many fungal secondary metabolites are used as medicines (e.g. penicillin or cyclosporine), or are of importance due to their toxicity in humans or animals (e.g. aflatoxins), or because they are virulence factors during crop plant infection (e.g. deoxynivalenol). Despite the vast potential encoded in ascomycete genomes, the vast majority of fungal secondary metabolites are yet to be discovered. This is in part due to our lack of understanding of how the production of these compounds is regulated. Moreover, even for the compounds we do know, we are just beginning to understand the complex interaction of external stimuli, signalling pathways, cellular and developmental biology that leads to their production. In this thesis, biological insights into these aspects of two fungal secondary metabolites, deoxynivalenol (DON) and fusatin are provided.In chapter II a high-throughput fluorescence activated cell sorting (FACS) based mutant screen was developed and the regulation of DON production in Fusarium graminearum was analysed. DON is an economically very important mycotoxin, as it is the most common contaminant of wheat, barley and corn worldwide. It is produced by plant pathogenic filamentous fungi of the Fusarium family, which can cause Fusarium head blight as well as Fusarium crown rot. During plant infection, DON is an important virulence factor and it can be toxic for humans and animals. The regulation of DON production is not yet fully understood. To identify regulators, a mutant screen was developed using FACS to allow high throughput screening, coupled with whole genome sequencing to identify mutations.Segregation analyses were performed to determine the mutation causing the phenotype.The mutant screen identified an adenylyl cyclase allele, which resulted in production of DON under normally repressive conditions. In addition, the mutant developed cellular structures associated with toxin production under repressive conditions. The levels of the fundamental second messenger cAMP, which adenylyl cyclases produce, were increased in the identified mutant, suggesting it might be a gain-of-function adenylyl cyclase mutant. This mutant screening methodology can be adapted to identify regulators of different secondary metabolites as well as to identify mutants overproducing yet unknown secondary metabolites. Abstract 2In chapter III the cell biology of DON production in Fusarium graminearum was elucidated with quantitative super resolution microscopy. During the last five years the first insights into the cell biology of DON production were gained, indicating complex cell biological mechanisms. During toxin production, F. graminearum develops endoplasmic reticulum proliferations, called toxisomes, at which two DON biosynthesis enzymes, TRI1 a...

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Genome Mining for Fungal Secondary Metabolic Gene Clusters

et al. 2015

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Fusarium graminearum Tri12p Influences Virulence to Wheat and Trichothecene Accumulation

Cited by 88 publications

References 53 publications

High-throughput FACS-based mutant screen identifies a gain-of-function allele of the Fusarium graminearum adenylyl cyclase causing deoxynivalenol over-production

High-throughput FACS-based mutant screen identifies a gain-of-function allele of the Fusarium graminearum adenylyl cyclase causing deoxynivalenol over-production

Regulation and cell biology of secondary metabolite production in Fusarium graminearum and Fusarium pseudograminearum

Genome Mining for Fungal Secondary Metabolic Gene Clusters

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