Colonization of maize silks by Fusarium graminearum, the causative organism of gibberella ear rot

2007 ◽  
Vol 85 (4) ◽  
pp. 369-376 ◽  
Author(s):  
S. Shea Miller ◽  
Lana M. Reid ◽  
Linda J. Harris
Keyword(s):  
Fusarium Graminearum ◽  
Fluorescent Protein ◽  
Fungal Growth ◽  
Infection Process ◽  
Gibberella Zeae ◽  
Causative Organism ◽  
Ear Rot ◽  
Wild Type ◽  
Gibberella Ear Rot ◽  
Maize Silks

One of the most economically important diseases of maize in Canada is gibberella ear rot caused by Fusarium graminearum Schwabe (teleomorph = Gibberella zeae (Schw.) Petch). Understanding how the fungus becomes established will help in developing effective strategies to reduce the incidence of this disease. This study investigates the infection process of F. graminearum on maize silks using both a wild-type F. graminearum as well as a strain transformed with a gene from jellyfish to constitutively express green fluorescent protein. Immature ears of maize were inoculated in the field with wild-type F. graminearum and harvested at specific times post infection, and the silks were stained with Chlorazol Black E for examination. In addition, uninoculated ears were excised, placed on water agar in large Petri dishes, and the silks inoculated with a suspension of macroconidia of the transformed fungus. The progress of fungal growth was then monitored using microscopy. Germination of conidia was observed 4–6 h after inoculation. A variable period of random growth often followed, after which some of the hyphae would grow in more or less straight lines down the silk towards the cob (rachis), and ultimately infect the developing kernels. Access to the cob occurred in 7–9 d in susceptible genotypes and 12–15 d in resistant genotypes. The fungus could penetrate the ovary directly through the silk attachment point or, when the silk was growing over other kernels, the fungus could traverse from the silk to colonize interkernel spaces. Entry into the cob was either through the rachis surface via exterior growth between kernels, or into the rachis via the pedicel.

scholarly journals Sphingolipid C-9 Methyltransferases Are Important for Growth and Virulence but Not for Sensitivity to Antifungal Plant Defensins in Fusarium graminearum

2008 ◽  
Vol 8 (2) ◽  
pp. 217-229 ◽  
Author(s):  
Vellaisamy Ramamoorthy ◽  
Edgar B. Cahoon ◽  
Mercy Thokala ◽  
Jagdeep Kaur ◽  
Jia Li ◽  
...  
Keyword(s):  
Fusarium Graminearum ◽  
Type Strain ◽  
Wild Type Strain ◽  
Fungal Growth ◽  
Wild Type ◽  
Knockout Mutant ◽  
Double Knockout ◽  
Disease Symptoms ◽  
Plant Defensins ◽  
Antifungal Action

ABSTRACT The C-9-methylated glucosylceramides (GlcCers) are sphingolipids unique to fungi. They play important roles in fungal growth and pathogenesis, and they act as receptors for some antifungal plant defensins. We have identified two genes, FgMT1 and FgMT2, that each encode a putative sphingolipid C-9 methyltransferase (C-9-MT) in the fungal pathogen Fusarium graminearum and complement a Pichia pastoris C-9-MT-null mutant. The ΔFgmt1 mutant produced C-9-methylated GlcCer like the wild-type strain, PH-1, whereas the ΔFgmt2 mutant produced 65 to 75% nonmethylated and 25 to 35% methylated GlcCer. No ΔFgmt1ΔFgmt2 double-knockout mutant producing only nonmethylated GlcCer could be recovered, suggesting that perhaps C-9-MTs are essential in this pathogen. This is in contrast to the nonessential nature of this enzyme in the unicellular fungus P. pastoris. The ΔFgmt2 mutant exhibited severe growth defects and produced abnormal conidia, while the ΔFgmt1 mutant grew like the wild-type strain, PH-1, under the conditions tested. The ΔFgmt2 mutant also exhibited drastically reduced disease symptoms in wheat and much-delayed disease symptoms in Arabidopsis thaliana. Surprisingly, the ΔFgmt2 mutant was less virulent on different host plants tested than the previously characterized ΔFggcs1 mutant, which lacks GlcCer synthase activity and produces no GlcCer at all. Moreover, the ΔFgmt1 and ΔFgmt2 mutants, as well as the P. pastoris strain in which the C-9-MT gene was deleted, retained sensitivity to the antifungal plant defensins MsDef1 and RsAFP2, indicating that the C-9 methyl group is not a critical structural feature of the GlcCer receptor required for the antifungal action of plant defensins.

scholarly journals Cotransformation of Trichoderma harzianum with β-Glucuronidase and Green Fluorescent Protein Genes Provides a Useful Tool for Monitoring Fungal Growth and Activity in Natural Soils

2000 ◽  
Vol 66 (2) ◽  
pp. 810-815 ◽  
Author(s):  
Yeoung-Seuk Bae ◽  
Guy R. Knudsen
Keyword(s):  
Green Fluorescent Protein ◽  
Trichoderma Harzianum ◽  
Fluorescent Protein ◽  
Fungal Growth ◽  
Marker Genes ◽  
Natural Soil ◽  
Nondestructive Monitoring ◽  
Wild Type ◽  
Glass Slides ◽  
Green Fluorescent

ABSTRACT Trichoderma harzianum was cotransformed with genes encoding green fluorescent protein (GFP), β-glucuronidase (GUS), and hygromycin B (hygB) resistance, using polyethylene glycol-mediated transformation. One cotransformant (ThzID1-M3) was mitotically stable for 6 months despite successive subculturing without selection pressure. ThzID1-M3 morphology was similar to that of the wild type; however, the mycelial growth rate on agar was reduced. ThzID1-M3 was formed into calcium alginate pellets and placed onto buried glass slides in a nonsterile soil, and its ability to grow, sporulate, and colonize sclerotia of Sclerotinia sclerotiorum was compared with that of the wild-type strain. Wild-type and transformant strains both colonized sclerotia at levels above those of indigenous Trichoderma spp. in untreated controls. There were no significant differences in colonization levels between wild-type and cotransformant strains; however, the presence of the GFP and GUS marker genes permitted differentiation of introducedTrichoderma from indigenous strains. GFP activity was a useful tool for nondestructive monitoring of the hyphal growth of the transformant in a natural soil. The green color of cotransformant hyphae was clearly visible with a UV epifluorescence microscope, while indigenous fungi in the same samples were barely visible. Green-fluorescing conidiophores and conidia were observed within the first 3 days of incubation in soil, and this was followed by the formation of terminal and intercalary chlamydospores and subsequent disintegration of older hyphal segments. Addition of 5-bromo-4-chloro-3-indolyl-β-d-glucuronic acid (X-Gluc) substrate to recovered glass slides confirmed the activity of GUS as well as GFP in soil. Our results suggest that cotransformation with GFP and GUS can provide a valuable tool for the detection and monitoring of specific strains of T. harzianum released into the soil.

scholarly journals Reduced Contamination by the Fusarium Mycotoxin Zearalenone in Maize Kernels through Genetic Modification with a Detoxification Gene

2007 ◽  
Vol 73 (5) ◽  
pp. 1622-1629 ◽  
Author(s):  
Tomoko Igawa ◽  
Naoko Takahashi-Ando ◽  
Noriyuki Ochiai ◽  
Shuichi Ohsato ◽  
Tsutomu Shimizu ◽  
...  
Keyword(s):  
Fluorescent Protein ◽  
Storage Period ◽  
Transgenic Maize ◽  
Ear Rot ◽  
Wild Type ◽  
Maize Seeds ◽  
Transgenic Seeds ◽  
In The Wild ◽  
Green Fluorescent ◽  
Maize Kernels

ABSTRACT Maize is subject to ear rot caused by toxigenic Aspergillus and Fusarium species, resulting in contamination with aflatoxins, fumonisins, trichothecenes, and zearalenone (ZEN). The trichothecene group and ZEN mycotoxins are produced by the cereal pathogen Fusarium graminearum. A transgenic detoxification system for the elimination of ZEN was previously developed using an egfp::zhd101 gene (gfzhd101), encoding an enhanced green fluorescent protein fused to a ZEN-degrading enzyme. In this study, we produced a transgenic maize line expressing an intact copy of gfzhd101 and examined the feasibility of transgene-mediated detoxification in the kernels. ZEN-degrading activity has been detected in transgenic kernels during seed maturation (for a period of 6 weeks after pollination). The level of detoxification activity was unaltered after an additional storage period of 16 weeks at 6°C. When the seeds were artificially contaminated by immersion in a ZEN solution for 48 h at 28°C, the total amount of the mycotoxin in the transgenic seeds was uniformly reduced to less than 1/10 of that in the wild type. The ZEN in the transgenic maize kernels was also efficiently decontaminated under conditions of lower water activity (aw) and temperature; e.g., 16.9 μg of ZEN was removed per gram of seed within 48 h at an aw of 0.90 at 20°C. F. graminearum infection assays demonstrated an absence of ZEN in the transgenic maize seeds, while the mycotoxin accumulated in wild-type kernels under the same conditions. Transgene-mediated detoxification may offer simple solutions to the problems of mycotoxin contamination in maize.

scholarly journals Fusarium graminearum Tri12p Influences Virulence to Wheat and Trichothecene Accumulation

2012 ◽  
Vol 25 (11) ◽  
pp. 1408-1418 ◽  
Author(s):  
Jon Menke ◽  
Yanhong Dong ◽  
H. Corby Kistler
Keyword(s):  
Fusarium Graminearum ◽  
Fluorescent Protein ◽  
Toxin Production ◽  
Major Facilitator Superfamily ◽  
Induction Medium ◽  
Wild Type ◽  
In Planta ◽  
Major Facilitator ◽  
Green Fluorescent ◽  
Self Protection

The gene Tri12 encodes a predicted major facilitator superfamily protein suggested to play a role in export of trichothecene mycotoxins produced by Fusarium spp. It is unclear, however, how the Tri12 protein (Tri12p) may influence trichothecene sensitivity and virulence of the wheat pathogen Fusarium graminearum. In this study, we establish a role for Tri12 in toxin accumulation and sensitivity as well as in pathogenicity toward wheat. Tri12 deletion mutants (tri12) are reduced in virulence and result in decreased trichothecene accumulation when inoculated on wheat compared with the wild-type strain or an ectopic mutant. Reduced radial growth of tri12 mutants on trichothecene biosynthesis induction medium was observed relative to the wild type and the ectopic strains. Diminished trichothecene accumulation was observed in liquid medium cultures inoculated with tri12 mutants. Wild-type fungal cells grown under conditions that induce trichothecene biosynthesis develop distinct subapical swelling and form large vacuoles. A strain expressing Tri12p linked to green fluorescent protein shows localization of the protein consistent with the plasma membrane. Our results indicate Tri12 plays a role in self-protection and influences toxin production and virulence of the fungus in planta.

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 The Stress-Activated Protein Kinase FgOS-2 Is a Key Regulator in the Life Cycle of the Cereal Pathogen Fusarium graminearum

2012 ◽  
Vol 25 (9) ◽  
pp. 1142-1156 ◽  
Author(s):  
Thuat Van Nguyen ◽  
Wilhelm Schäfer ◽  
Jörg Bormann
Keyword(s):  
Protein Kinase ◽  
Fusarium Graminearum ◽  
Fluorescent Protein ◽  
Microscopic Analysis ◽  
Deletion Mutants ◽  
Wild Type ◽  
In Planta ◽  
Head Blight ◽  
Food And Feed

Fusarium graminearum is one of the most destructive pathogens of cereals and a threat to food and feed production worldwide. It is an ascomycetous plant pathogen and the causal agent of Fusarium head blight disease in small grain cereals and of cob rot disease in maize. Infection with F. graminearum leads to yield losses and mycotoxin contamination. Zearalenone (ZEA) and deoxynivalenol (DON) are hazardous mycotoxins; the latter is necessary for virulence toward wheat. Deletion mutants of the F. graminearum orthologue of the Saccharomyces cerevisiae Hog1 stress-activated protein kinase, FgOS-2 (ΔFgOS-2), showed drastically reduced in planta DON and ZEA production. However, ΔFgOS-2 produced even more DON than the wild type under in vitro conditions, whereas ZEA production was similar to that of the wild type. These deletion strains are dramatically reduced in pathogenicity toward maize and wheat. We constitutively expressed the fluorescent protein dsRed in the deletion strains and the wild type. Microscopic analysis revealed that ΔFgOS-2 is unable to reach the rachis node at the base of wheat spikelets. During vegetative growth, ΔFgOS-2 strains exhibit increased resistance against the phenylpyrrole fludioxonil. Growth of mutant colonies on agar plates supplemented with NaCl is reduced but conidia formation remained unchanged. However, germination of mutant conidia on osmotic media is severely impaired. Germ tubes are swollen and contain multiple nuclei. The deletion mutants completely fail to produce perithecia and ascospores. Furthermore, FgOS-2 also plays a role in reactive oxygen species (ROS)-related signaling. The transcription and activity of fungal catalases is modulated by FgOS-2. Among the genes regulated by FgOS-2, we found a putative calcium-dependent NADPH-oxidase (noxC) and the transcriptional regulator of ROS metabolism, atf1. The present study describes new aspects of stress-activated protein kinase signaling in F. graminearum.

scholarly journals Chlorogenic Acid and Maize Ear Rot Resistance: A Dynamic Study Investigating Fusarium graminearum Development, Deoxynivalenol Production, and Phenolic Acid Accumulation

2012 ◽  
Vol 25 (12) ◽  
pp. 1605-1616 ◽  
Author(s):  
Vessela Atanasova-Penichon ◽  
Sebastien Pons ◽  
Laetitia Pinson-Gadais ◽  
Adeline Picot ◽  
Gisèle Marchegay ◽  
...  
Keyword(s):  
Chlorogenic Acid ◽  
Fusarium Graminearum ◽  
Phenolic Acid ◽  
Quantitative Polymerase Chain Reaction ◽  
Resistant Variety ◽  
Ear Rot ◽  
In Planta ◽  
Gibberella Ear Rot ◽  
Moderately Resistant ◽  
Maize Kernels

Fusarium graminearum is the causal agent of Gibberella ear rot and produces trichothecene mycotoxins. Basic questions remain unanswered regarding the kernel stages associated with trichothecene biosynthesis and the kernel metabolites potentially involved in the regulation of trichothecene production in planta. In a two-year field study, F. graminearum growth, trichothecene accumulation, and phenolic acid composition were monitored in developing maize kernels of a susceptible and a moderately resistant variety using quantitative polymerase chain reaction and liquid chromatography coupled with photodiode array or mass spectrometry detection. Infection started as early as the blister stage and proceeded slowly until the dough stage. Then, a peak of trichothecene accumulation occurred and infection progressed exponentially until the final harvest time. Both F. graminearum growth and trichothecene production were drastically reduced in the moderately resistant variety. We found that chlorogenic acid is more abundant in the moderately resistant variety, with levels spiking in the earliest kernel stages induced by Fusarium infection. This is the first report that precisely describes the kernel stage associated with the initiation of trichothecene production and provides in planta evidence that chlorogenic acid may play a role in maize resistance to Gibberella ear rot and trichothecene accumulation.

scholarly journals Reaction of maize hybrids to ear rot caused by Fusarium graminearum Schwabe

2017 ◽  
Vol 38 (SI 2 - 6th Conf EFPP 2002) ◽  
pp. 569-571 ◽  
Author(s):  
M. Pastirčák ◽  
M. Lemmens ◽  
A. Šrobárová
Keyword(s):  
Fusarium Graminearum ◽  
Gibberella Zeae ◽  
Conidial Suspension ◽  
Fusarium Ear Rot ◽  
Ear Rot ◽  
Maize Hybrids ◽  
Inoculation Techniques ◽  
Maize Resistance ◽  
The Way

Ear rot caused by Fusarium graminearum Schwabe (teleomorph stage: Gibberella zeae (Schwein.) Petch) is a destructive disease of maize. In our experiment we tested twenty maize hybrids. Two inoculation techniques differing in the way of application of a macro-conidial suspension, were evaluated for their effectiveness in assessing maize resistance to ear rot. Based on the results of one season, highly significant differences in sensitivity to Fusarium ear rot between genotypes for all variants under mist irrigation and without mist irrigation, were detected.

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.

Evaluation of Fusarium graminearum inoculation methods in maize ears and hybrid reaction to Gibberella ear rot under southern Brazilian environmental conditions

2015 ◽  
Vol 144 (1) ◽  
pp. 45-53 ◽  
Author(s):  
Francine R. Nerbass ◽  
Ricardo T. Casa ◽  
Paulo Roberto Kuhnem ◽  
Amauri Bogo ◽  
Luis Sangoi ◽  
...  
Keyword(s):  
Fusarium Graminearum ◽  
Environmental Conditions ◽  
Ear Rot ◽  
Gibberella Ear Rot ◽  
Inoculation Methods
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