scholarly journals Functional and Comparative Bioinformatic Analysis of Expressed Genes from Wheat Spikes Infected with Fusarium graminearum

2002 ◽  
Vol 15 (5) ◽  
pp. 445-455 ◽  
Author(s):  
Warren M. Kruger ◽  
Clara Pritsch ◽  
Shiaoman Chao ◽  
Gary J. Muehlbauer
Keyword(s):  
Cdna Library ◽  
Fusarium Graminearum ◽  
Plant Cell Wall ◽  
Sequence Similarity ◽  
Amino Acid Sequence Similarity ◽  
Biotic And Abiotic Stress ◽  
Head Blight ◽  
Stress Related Genes ◽  
Fungal Genes ◽  
Sumai 3

Fusarium head blight, caused by the fungus Fusarium graminearum, is a major disease on wheat (Triticum aestivum L.). Expressed sequence tags (ESTs) were used to identify genes expressed during the wheat-F. graminearum interaction. We generated 4,838 ESTs from a cDNA library prepared from spikes of the partially resistant cultivar Sumai 3 infected with F. graminearum. These ESTs were composed of 2,831 singlet (single-copy transcripts) and 715 contigs (multiple-copy transcripts) for a total of 3,546 non-redundant sequences. Four sets of nonredundant sequences were identified. One set contains numerous, common biotic and abiotic stress-related genes. Many of these stress-related genes were represented by multiple ESTs, indicating that they are abundantly expressed. A second set comprised 16 nonredundant sequences from F. graminearum that may be required for pathogenicity. A subset of these fungal genes encodes proteins associated with plant cell wall degradation. A third set of 326 nonredundant sequences had no DNA or amino acid sequence similarity to almost 1 million plant and over 7 million animal sequences in dbEST (as of 22 June 2001). Thus, these 326 nonredundant sequences have only been found in our F. graminearum-infected ‘Sumai 3’ cDNA library. A fourth set of 29 nonredundant sequences was found in our F. graminearum-infected wheat and another plant-pathogen interaction cDNA library. Some of these sequences encode proteins that may act in establishing various plant-fungal interactions.

scholarly journals Tri1 in Fusarium graminearum Encodes a P450 Oxygenase

2004 ◽  
Vol 70 (4) ◽  
pp. 2044-2051 ◽  
Author(s):  
S. P. McCormick ◽  
L. J. Harris ◽  
N. J. Alexander ◽  
T. Ouellet ◽  
A. Saparno ◽  
...  
Keyword(s):  
Cytochrome P450 ◽  
Fusarium Graminearum ◽  
Sequence Similarity ◽  
Gibberella Zeae ◽  
Cdna Libraries ◽  
Amino Acid Sequence Similarity ◽  
Ear Rot ◽  
In Planta ◽  
Head Blight ◽  
Asexual State

ABSTRACT Gibberella zeae (asexual state Fusarium graminearum) is a major causal agent of wheat head blight and maize ear rot in North America and is responsible for contamination of grain with deoxynivalenol and related trichothecene mycotoxins. To identify additional trichothecene biosynthetic genes, cDNA libraries were prepared from fungal cultures under trichothecene-inducing conditions in culture and in planta. A gene designated LH1 that was highly expressed under these conditions exhibited only moderate (59%) similarity to known trichothecene biosynthetic cytochrome P450s. To determine the function of LH1, gene disruptants were produced and assessed for trichothecene production. Gene disruptants no longer produced 15-acetyldeoxynivalenol, which is oxygenated at carbon 7 (C-7) and C-8, but rather accumulated calonectrin and 3-deacetylcalonectrin, which are not oxygenated at either C-7 or C-8. These results indicate that gene LH1 encodes a cytochrome P450 responsible for oxygenation at one or both of these positions. Despite the relatively low level of DNA and amino acid sequence similarity between the two genes, LH1 from G. zeae is the probable homologue of Tri1, which encodes a cytochrome P450 required for C-8 oxygenation in F. sporotrichioides.

scholarly journals Novel Genes of Fusarium graminearum That Negatively Regulate Deoxynivalenol Production and Virulence

2009 ◽  
Vol 22 (12) ◽  
pp. 1588-1600 ◽  
Author(s):  
Donald M. Gardiner ◽  
Kemal Kazan ◽  
John M. Manners
Keyword(s):  
Gene Expression ◽  
Fusarium Graminearum ◽  
Global Analysis ◽  
Gene Clusters ◽  
Head Blight ◽  
Knock Out ◽  
Serious Disease ◽  
Amine Compounds ◽  
Fungal Genes ◽  
Fungal Gene Expression

Fusarium head blight of wheat, caused by Fusarium graminearum, is a serious disease resulting in both reduced yields and contamination of grain with trichothecene toxins, with severe consequences for mammalian health. Recently, we have identified several related amine compounds such as agmatine and putrescine that promote the production of high levels of trichothecene toxins, such as deoxynivalenol (DON), in culture by F. graminearum and F. sporotrichioides. Here, a global analysis of fungal gene expression using the Affymetrix Fusarium GeneChip during culture under DON-inducing conditions compared with noninducing conditions is reported. Agmatine differentially regulated a large number of fungal genes, including both known and previously uncharacterized putative secondary metabolite biosynthetic gene clusters. In silico prediction of binding sites for the transcriptional regulator (TRI6) controlling TRI gene expression and gene expression analysis in a TRI6 mutant of F. graminearum showed that three of the differentially regulated genes were under the control of TRI6. Gene knock-out mutations of two of these genes resulted in mutants with massively increased production of DON and increased aggressiveness toward wheat. Our results not only identify a novel mechanism of negative regulation of DON production and virulence in F. graminearum but also point out the potential of this pathogen to evolve with an ability to produce massively increased amounts of toxins and increased virulence.

Induction of wheat defense and stress-related genes in response to Fusarium graminearum

Genome ◽  
2005 ◽  
Vol 48 (1) ◽  
pp. 29-40 ◽  
Author(s):  
Lingrang Kong ◽  
Joseph M Anderson ◽  
Herbert W Ohm
Keyword(s):  
Fusarium Head Blight ◽  
Real Time ◽  
Cdna Library ◽  
Fusarium Graminearum ◽  
Cdna Clone ◽  
Subtractive Hybridization ◽  
Suppressive Subtractive Hybridization ◽  
Nucleotide Sequence Analysis ◽  
Head Blight ◽  
Resistance Response

Fusarium head blight (FHB), caused by species of the fungus Fusarium, is a worldwide disease of wheat (Triticum aestivum L.). The Chinese T. aestivum 'Ning7840' is one of few wheat cultivars with resistance to FHB. To identify differentially expressed genes corresponding to FHB resistance, a cDNA library was constructed using pooled mRNA isolated from glumes of 'Ning7840' harvested at 2, 6, 12, 24, 36, 72, and 96 h after inoculation (hai) with a conidia spore suspension of Fusarium graminearum. Suppressive subtractive hybridization (SSH) cDNA subtraction was carried out using pooled glume mRNAs from the tester and the control. The cDNA library was differentially screened using the forward subtracted cDNAs and the reverse subtracted cDNAs as probes. Twenty-four clones with significant matches to either plant (16 sequences) or fungal (8 sequences) genes were isolated based on their specific hybridization with forward subtracted cDNA and not reverse subtracted cDNA. Six putative defense-related genes were confirmed by real-time quantitative reverse-transcriptase PCR. Many-fold higher induction of three clones (A3F8, B10H1, and B11H3) in the resistant genotypes compared with susceptible genotypes indicates a putative role in the resistance response to Fusarium graminearum. Transcript accumulations of P450, chitinase (Chi1), and one unknown gene (clone B8Q9) in both resistant and susceptible genotypes suggest an involvement in a generalized resistance response to F. graminearum. Nucleotide sequence analysis showed that cDNA clone A4C6 encodes a cytochrome P450 gene (CYP709C3v2), including 14 N-terminal amino acids that have a membrane-associated helical motif. Other domains characteristic of eukaryotic P450 are also present in CYP709C3v2. The deduced polypeptide of cDNA clone B2H2 encodes an acidic isoform of class I chitinase containing a 960-bp coding region. Southern hybridization using aneuploid lines of T. aestivum 'Chinese Spring' indicated that CYP709C3v2 was located on the short arm of chromosomes 2B and 2D.Key words: Fusarium head blight (FHB), suppressive subtractive hybridization, defense response, real-time quantitative RT-PCR.

scholarly journals The Transcriptome of Fusarium graminearum During the Infection of Wheat

2011 ◽  
Vol 24 (9) ◽  
pp. 995-1000 ◽  
Author(s):  
Erik Lysøe ◽  
Kye-Yong Seong ◽  
H. Corby Kistler
Keyword(s):  
Fusarium Graminearum ◽  
Global Gene Expression ◽  
Infection Process ◽  
Complete Medium ◽  
Post Inoculation ◽  
Head Blight ◽  
Single Nucleotide ◽  
Evolutionary Selection ◽  
Blight Disease ◽  
Fungal Genes

Fusarium graminearum causes head blight disease in wheat and barley. To help understand the infection process on wheat, we studied global gene expression of F. graminearum in a time series from 24 to 196 h after inoculation, compared with a noninoculated control. The infection was rapid and, after 48 h, over 4,000 fungal genes were expressed. The number of genes expressed increased over time up to 96 h (>8,000 genes), and then declined at the 144- and 192-h post-inoculation time points. After subtraction of genes found expressed on complete medium, during carbon or nitrogen starvation, and on barley, only 355 were found exclusively expressed in wheat, mostly genes with unknown function (72.6%). These genes were mainly found in single-nucleotide polymorphism-enriched islands on the chromosomes, suggesting a higher evolutionary selection pressure. The annotated genes were enriched in functional groups predicted to be involved in allantoin and allantoate transport, detoxification, nitrogen, sulfur and selenium metabolism, secondary metabolism, carbohydrate metabolism, and degradation of polysaccharides and ester compounds. Several putative secreted virulence factors were also found expressed in wheat.

scholarly journals Fungal gene expression during ectomycorrhiza formation

1995 ◽  
Vol 73 (S1) ◽  
pp. 541-547 ◽  
Author(s):  
F. Martin ◽  
P. Laurent ◽  
D. de Carvalho ◽  
T. Burgess ◽  
P. Murphy ◽  
...  
Keyword(s):  
Cell Walls ◽  
Molecular Mechanisms ◽  
Sequence Similarity ◽  
Polyacrylamide Gel Electrophoresis ◽  
Pisolithus Tinctorius ◽  
Amino Acid Sequence Similarity ◽  
Cell Fractionation ◽  
Fungal Cell ◽  
Temporal Expressions ◽  
Fungal Genes

Ectomycorrhiza development involves the differentiation of structurally specialized fungal tissues (e.g., mantle and Hartig net) and an interface between symbionts. Polypeptides presenting a preferential, up-, or down-regulated synthesis have been characterized in several developing ectomycorrhizal associations. Their spatial and temporal expressions have been characterized by cell fractionation, two-dimensional polyacrylamide gel electrophoresis, and immunochemical assays in the Eucalyptus spp. – Pisolithus tinctorius mycorrhizas. These studies have emphasized the importance of fungal cell wall polypeptides during the early stages of the ectomycorrhizal interaction. The increased synthesis of 30- to 32-kDa acidic polypeptides, together with the decreased accumulation of a prominent 95-kDa mannoprotein provided evidence for major alterations of Pisolithus tinctorius cell walls during mycorrhiza formation. Differential cDNA library screening and shotgun cDNA sequencing were used to clone symbiosis-regulated fungal genes. Several abundant transcripts showed a significant amino acid sequence similarity to a family of secreted morphogenetic fungal proteins, the so-called hydrophobic. In P. tinctorius, the content of hydrophobin transcripts is high in aerial hyphae and during the ectomycorrhizal sheath formation. Alteration of cell walls and the extracellular matrix is therefore a key event in the ectomycorrhiza development. An understanding of the molecular mechanisms that underlies the temporal and spatial control of genes and proteins involved in the development of the symbiotic interface is now within reach, as more sophisticated techniques of molecular and genetic analysis are applied to the mycorrhizal interactions. Key words: cell walls, ectomycorrhiza, ectomycorrhizins, fungal development, hydrophobins, symbiosis-regulated polypeptides.

scholarly journals Transcript Abundance Responses of Resistance Pathways of Arabidopsis thaliana to Deoxynivalenol

2017 ◽  
Vol 2 (3) ◽  
pp. 154-161
Author(s):  
Jiazheng Yuan ◽  
Michelle Zhu ◽  
Khalid Meksem ◽  
Matt Geisler ◽  
Patrick Hart ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Fusarium Graminearum ◽  
Transcript Abundance ◽  
Triticum Aestivum L ◽  
Gibberella Zeae ◽  
Ear Rot ◽  
Head Blight ◽  
Soybean Roots ◽  
Stress Related Genes ◽  
Expressed Sequence

Mycotoxin deoxynivalenol (DON), produced by Gibberella zeae (Schwein.) Petch (teleomorph of Fusarium graminearum Schwabe) was known to be both a virulence factor in the pathogenesis of Triticum aestivum L. (wheat) and an inhibitor of Arabidopsis thaliana L. seed germination. Fusarium graminearum causes both Gibberella ear rot in maize (Zea mays L.) and Fusarium head blight (FHB) in wheat and barley. Arabidopsis thaliana was also a host for the related root rot pathogen F. virguliforme Aoki. A. thaliana seedling growth was reduced by the pathogen in a proportional response to increasing spore concentrations. Here, the changes in transcript abundances corresponding to 10,560 A. thaliana expressed sequence tags (ESTs) was compared with changes in 192 known plant defense and biotic/abiotic stress related genes in soybean roots after infestation with F. virguliforme. A parallel comparison with a set of resistance pathways involved in response to the DON toxicity in A. thaliana was performed. A. thaliana data was obtained from the AFGC depository. The variations of transcript abundances in Arabidopsis and soybean treated with pathogen suggest that both plants respond to the pathogen mainly by common, possibly global responses with some specific secondary metabolic pathways involved in defense. In contrast, DON toxin appeared to impact central metabolisms in Arabidopsis plants with significant alterations ranging from the protein metabolism to redox production. Several new putative resistance pathways involved in responding to both pathogen and DON infestation in soybean and A. thaliana were identified.

scholarly journals Fungal Development and Induction of Defense Response Genes During Early Infection of Wheat Spikes by Fusarium graminearum

2000 ◽  
Vol 13 (2) ◽  
pp. 159-169 ◽  
Author(s):  
Clara Pritsch ◽  
Gary J. Muehlbauer ◽  
William R. Bushnell ◽  
David A. Somers ◽  
Carroll P. Vance
Keyword(s):  
Fusarium Graminearum ◽  
Defense Response ◽  
Expression Patterns ◽  
Economic Losses ◽  
Temporal Expression ◽  
Head Blight ◽  
Transcript Accumulation ◽  
Genes Encoding ◽  
Defense Response Genes ◽  
Sumai 3

Fusarium head blight (FHB) of wheat is a crippling disease that causes severe economic losses in many of the wheat-growing regions of the world. Temporal patterns of fungus development and transcript accumulation of defense response genes were studied in Fusarium graminearum-inoculated wheat spikes within the first 48 to 76 h after inoculation (hai). Microscopy of inoculated glumes revealed that the fungus appeared to penetrate through stomata, exhibited subcuticular growth along stomatal rows, colonized glume parenchyma cells, and sporulated within 48 to 76 hai. No major differences in the timing of these events were found between Sumai 3 (resistant) and Wheaton (susceptible) genotypes. In complementary experiments, RNA was extracted from spikes at several time intervals up to 48 hai and temporal expression patterns were determined for defense response genes encoding peroxidase, PR-1, PR-2 (β-1,3-glucanase), PR-3 (chitinase), PR-4, and PR-5 (thaumatin-like protein). In both genotypes, transcripts for the six defense response genes accumulated as early as 6 to 12 hai during F. graminearum infection and peaked at 36 to 48 hai. Greater and earlier PR-4 and PR-5 transcript accumulation was observed in Sumai 3, compared with Wheaton. Our results show that the timing of defense response gene induction is correlated with F. graminearum infection.

scholarly journals Genomic analysis of host–pathogen interaction between Fusarium graminearum and wheat during early stages of disease development

Microbiology ◽  
2006 ◽  
Vol 152 (6) ◽  
pp. 1877-1890 ◽  
Author(s):  
Rubella S. Goswami ◽  
Jin-Rong Xu ◽  
Frances Trail ◽  
Karen Hilburn ◽  
H. Corby Kistler
Keyword(s):  
Fusarium Graminearum ◽  
Response Regulator ◽  
Gene Prediction ◽  
Genomic Analysis ◽  
Bioinformatic Analysis ◽  
Gene Replacement ◽  
Cdna Libraries ◽  
Disease Development ◽  
Head Blight ◽  
Fungal Genes

Fusarium graminearum strains responsible for causing the plant disease Fusarium head blight vary greatly in their ability to cause disease and produce mycotoxins on wheat. With the goal of understanding fungal gene expression related to pathogenicity, three cDNA libraries were created by suppression subtractive hybridization using wheat heads inoculated with a highly aggressive strain and either water or a less aggressive strain of this pathogen. Eighty-four fungal genes expressed during initial disease development were identified. The probable functions of 49 of these genes could be inferred by bioinformatic analysis. Thirty-five ESTs had no known homologues in current databases and were not identified by ab initio gene prediction methods. These ESTs from infected wheat heads probably represent F. graminearum genes that previously were not annotated. Four genes represented in one of these libraries were selected for targeted gene replacement, leading to the characterization of a two-component response regulator homologue involved in pathogenicity of the fungus. The mutants for this gene showed reduced sporulation and delayed spread of Fusarium head blight on wheat.

scholarly journals Screening of Durum Wheat (Triticum turgidum L. subsp. durum (Desf.) Husn.) Italian Cultivars for Susceptibility to Fusarium Head Blight Incited by Fusarium graminearum

Plants ◽  
2020 ◽  
Vol 10 (1) ◽  
pp. 68
Author(s):  
Gaetano Bentivenga ◽  
Alfio Spina ◽  
Karim Ammar ◽  
Maria Allegra ◽  
Santa Olga Cacciola
Keyword(s):  
Durum Wheat ◽  
Fusarium Head Blight ◽  
Fusarium Graminearum ◽  
Triticum Turgidum ◽  
Disease Incidence ◽  
Morphological Characteristics ◽  
Pcr Amplification ◽  
Conidial Suspension ◽  
Head Blight ◽  
Italian Origin

In 2009, a set of 35 cultivars of durum wheat (Triticum turgidum L. subsp. durum (Desf.) Husn.) of Italian origin was screened for fusarium head blight (FHB) susceptibility at CIMMYT (Mexico) and in the 2019–20 cropping season, 16 of these cultivars, which had been included in the Italian National Plant Variety Register, were tested again in southern and northern Italy. Wheat cultivars were artificially inoculated during anthesis with a conidial suspension of Fusarium graminearum sensu lato using a standard spray inoculation method. Inoculum was a mixture of mono-conidial isolates sourced in the same areas where the trials were performed. Isolates had been characterized on the basis of morphological characteristics and by DNA PCR amplification using a specific primer set and then selected for their virulence and ability to produce mycotoxins. The susceptibility to FHB was rated on the basis of the disease severity, disease incidence and FHB index. Almost all of the tested cultivars were susceptible or very susceptible to FHB with the only exception of “Duprì”, “Tiziana” and “Dylan” which proved to be moderately susceptible. The susceptibility to FHB was inversely correlated with the plant height and flowering biology, the tall and the late heading cultivars being less susceptible.

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