Temperature-responded biological fitness of carbendazim-resistance Fusarium graminearum mutants conferring the F167Y, E198K and E198L substitutions

Plant Disease ◽  
2021 ◽  
Author(s):  
Yingfan Wang ◽  
Yi-ping Hou ◽  
XueWei Mao ◽  
fuyu Liu ◽  
Mingguo Zhou
Keyword(s):  
Low Temperature ◽  
Fusarium Graminearum ◽  
Hyphal Growth ◽  
Atmospheric Temperature ◽  
Cross Resistance ◽  
Wild Type ◽  
Head Blight ◽  
Flower Stage ◽  
Effects Of Temperature ◽  
Resistance Mutants

Understanding the effects of temperature on Fusarium graminearum infection can provide the theoretical guidance for chemical control of Fusarium head blight (FHB)．Here, we evaluated the effects of various temperatures on biological fitness development of wild-type sensitive strain 2021 and carbendazim-resistance mutants conferring β2-tubulin substitutions F167Y, E198K and E198L. The results showed that mycelial growth and conidiation of four strains increased with the increase of the temperature between 10°C and 25°C. Conidia of F167Y displayed strong adaptability to low temperature. The virulence of the four strains was largely similar at the same temperature, showing an upward trend between 10°C and 25°C. At 10°C, the hyphal growth of all strains was significantly inhibited, and metabolism slowed down and the accumulation of secondary metabolites decreased. Subsequently, the production of deoxynivalenol (DON) and its intermediate, pyruvate and aurofusarin decreased at low temperature, and the expression of DON biosynthesis-related genes Tri5, FgPK and AUR decreased accordingly. At the same temperature, the aurofusarin production of the strains F167Y and E198K was higher than that of strains 2021 and E198L. The contents of DON and pyruvic acid in carbendazim-resistance mutants were higher than that of the wild-type strain 2021. The sensitivity of four strains to different fungicides changed at various temperatures. The sensitivity to most fungicides increased with the temperature decreasing. The carbendazim-resistance mutants appeared positive cross-resistance with other benzimidazoles. But there was no cross-resistance to pyraclostrobin and azoles. These results would direct us to use fungicide preventing the infection of F. graminearum with changeable atmospheric temperature at wheat flower stage.

scholarly journals A Natural Mutation Involving both Pathogenicity and Perithecium Formation in the Fusarium graminearum Species Complex

2016 ◽  
Vol 6 (12) ◽  
pp. 3883-3892 ◽  
Author(s):  
Haruhisha Suga ◽  
Koji Kageyama ◽  
Masafumi Shimizu ◽  
Misturo Hyakumachi
Keyword(s):  
Mutant Strain ◽  
Fusarium Graminearum ◽  
Type Strain ◽  
Species Complex ◽  
Wild Type Strain ◽  
Genetic Locus ◽  
Chromosome 2 ◽  
Wild Type ◽  
Head Blight ◽  
Fusarium Graminearum Species Complex

Abstract Members of the Fusarium graminearum species complex (Fg complex or FGSC) are the primary pathogens causing Fusarium head blight in wheat and barley worldwide. A natural pathogenicity mutant (strain 0225022) was found in a sample of the Fg complex collected in Japan. The mutant strain did not induce symptoms in wheat spikes beyond the point of inoculation, and did not form perithecia. No segregation of phenotypic deficiencies occurred in the progenies of a cross between the mutant and a fully pathogenic wild-type strain, which suggested that a single genetic locus controlled both traits. The locus was mapped to chromosome 2 by using sequence-tagged markers; and a deletion of ∼3 kb was detected in the mapped region of the mutant strain. The wild-type strain contains the FGSG_02810 gene, encoding a putative glycosylphosphatidylinositol anchor protein, in this region. The contribution of FGSG_02810 to pathogenicity and perithecium formation was confirmed by complementation in the mutant strain using gene transfer, and by gene disruption in the wild-type strain.

scholarly journals Enhanced Resistance to Fusarium graminearum in Transgenic Arabidopsis Plants Expressing a Modified Plant Thionin

2020 ◽  
Vol 110 (5) ◽  
pp. 1056-1066 ◽  
Author(s):  
Guixia Hao ◽  
Matthew G. Bakker ◽  
Hye-Seon Kim
Keyword(s):  
Transgenic Plants ◽  
Fusarium Graminearum ◽  
Transgenic Arabidopsis ◽  
Hyphal Growth ◽  
Marker Genes ◽  
Head Blight ◽  
Transgenic Expression ◽  
Transgenic Arabidopsis Plants ◽  
Enhanced Resistance ◽  
And Control

The fungal pathogen Fusarium graminearum causes Fusarium head blight (FHB) on wheat, barley, and other grains. FHB results in yield reductions and contaminates grain with trichothecene mycotoxins, which threaten food safety and food security. Innovative mechanisms for controlling FHB are urgently needed. We have previously shown that transgenic tobacco and citrus plants expressing a modified thionin (Mthionin) exhibited enhanced resistance toward several bacterial pathogens. The aim of this study was to investigate whether overexpression of Mthionin could be similarly efficacious against F. graminearum, and whether transgenic expression of Mthionin impacts the plant microbiome. Transgenic Arabidopsis plants expressing Mthionin were generated and confirmed. When challenged with F. graminearum, Mthionin-expressing plants showed less disease and fungal biomass in both leaves and inflorescences compared with control plants. When infiltrated into leaves, macroconidia of F. graminearum germinated at lower rates and produced less hyphal growth in Arabidopsis leaves expressing Mthionin. Moreover, marker genes related to defense signaling pathways were expressed at significantly higher levels after F. graminearum infection in Mthionin transgenic Arabidopsis plants. However, Mthionin expression did not appreciably alter the overall microbiome associated with transgenic plants grown under controlled conditions; across leaves and roots of Mthionin-expressing and control transgenic plants, only a few bacterial and fungal taxa differed, and differences between Mthionin transformants were of similar magnitude compared with control plants. In sum, our data indicate that Mthionin is a promising candidate to produce transgenic crops for reducing FHB severity and ultimately mycotoxin contamination.

scholarly journals A sterol C-14 reductase encoded by FgERG24B is responsible for the intrinsic resistance of Fusarium graminearum to amine fungicides

Microbiology ◽  
2011 ◽  
Vol 157 (6) ◽  
pp. 1665-1675 ◽  
Author(s):  
Xin Liu ◽  
Jing Fu ◽  
Yingzi Yun ◽  
Yanni Yin ◽  
Zhonghua Ma
Keyword(s):  
Fusarium Graminearum ◽  
Type Strain ◽  
Wild Type Strain ◽  
Ergosterol Biosynthesis ◽  
Deletion Mutants ◽  
Wild Type ◽  
Head Blight ◽  
Intrinsic Resistance ◽  
In The Wild ◽  
Increased Sensitivity

Fusarium graminearum, the causal agent of wheat head blight, shows intrinsic resistance to amine fungicides. It is commonly accepted that the amines target sterol C-14 reductase and sterol Δ8–Δ7 isomerase of ergosterol biosynthesis, encoded by the genes ERG24 and ERG2, respectively. Analysis of the genome sequence of F. graminearum revealed that the fungus contains two paralogous FgERG24 genes (FgERG24A and FgERG24B), which are homologous to the ERG24 of Saccharomyces cerevisiae. In this study, we disrupted FgERG24A and FgERG24B in F. graminearum. Compared to the wild-type strain HN9-1, FgERG24A and FgERG24B deletion mutants did not show recognizable phenotypic changes in mycelial growth on potato dextrose agar or in virulence on wheat heads. HPLC analysis showed that the amount of ergosterol in FgERG24A or FgERG24B deletion mutants was not significantly different from that in the wild-type strain. These results indicate that neither of the two genes is essential for growth, pathogenicity or ergosterol biosynthesis in F. graminearum. FgERG24B deletion mutants exhibited significantly increased sensitivity to amine fungicides, including tridemorph, fenpropidin and spiroxamine, but not to non-amine fungicides. In contrast, FgERG24A deletion mutants did not show changed sensitivity to any amine tested. The resistance of the FgERG24B deletion mutant to amines was restored by genetic complementation of the mutant with wild-type FgERG24B. These results indicate that FgERG24B controls the intrinsic resistance of F. graminearum to amines. The finding of this study provides new insights into amine resistance in filamentous fungi.

scholarly journals Effects of Temperature and Moisture on Development of Fusarium graminearum Perithecia in Maize Stalk Residues

2015 ◽  
Vol 82 (1) ◽  
pp. 184-191 ◽  
Author(s):  
Valentina Manstretta ◽  
Vittorio Rossi
Keyword(s):  
Fusarium Graminearum ◽  
Crop Residues ◽  
Head Blight ◽  
Dispersal Patterns ◽  
Content Type ◽  
Predominant Component ◽  
Effects Of Temperature ◽  
Ascospore Production ◽  
Maize Stalks ◽  
Over Time

ABSTRACTFusarium graminearumis the predominant component of the Fusarium head blight complex of wheat.F. graminearumascospores, which initiate head infection, mature in perithecia on crop residues and become airborne. The effects of temperature (T) and moisture on perithecium production and maturation and on ascospore production on maize stalk residues were determined. In the laboratory, perithecia were produced at temperatures between 5 and 30°C (the optimum was 21.7°C) but matured only at 20 and 25°C. Perithecia were produced when relative humidity (RH) was ≥75% but matured only when RH was ≥85%; perithecium production and maturation increased with RH. Equations describing perithecium production and maturation over time as a function ofTand RH (R2> 0.96) were developed. Maize stalks were also placed outdoors on three substrates: a grass lawn exposed to rain; a constantly wet, spongelike foam exposed to rain; and a grass lawn protected from rain. No perithecia were produced on stalks protected from rain. Perithecium production and maturation were significantly higher on the constantly wet foam than on the intermittently wet lawn (both exposed to rain). Ascospore numbers but not their dispersal patterns were also affected by the substrate.

scholarly journals The Semidwarfing Alleles Rht-D1b and Rht-B1b Show Marked Differences in Their Associations with Anther-Retention in Wheat Heads and with Fusarium Head Blight Susceptibility

2016 ◽  
Vol 106 (12) ◽  
pp. 1544-1552 ◽  
Author(s):  
Maria Buerstmayr ◽  
Hermann Buerstmayr
Keyword(s):  
Fusarium Head Blight ◽  
Plant Height ◽  
Fusarium Graminearum ◽  
Hyphal Growth ◽  
Field Trials ◽  
Wheat Breeding ◽  
Head Blight ◽  
Backcross Population ◽  
Reduced Height ◽  
Haploid Population

The semidwarfing alleles reduced height (Rht)-D1b and Rht-B1b are widely deployed in wheat breeding. Both alleles have similar effects on plant height but differ in their effect on Fusarium head blight (FHB) severity. A double-haploid population and a backcross population, segregating for Rht-B1a/Rht-B1b and Rht-D1a/Rht-D1b, were evaluated for FHB severity, plant height, and anther retention in field trials in three consecutive years. The semidwarfing alleles reduced plant height and increased the proportion of retained anthers. Reduced plant height and a high proportion of retained anthers were associated with increased FHB severity. The Rht-D1b allele had a significantly greater impact on anther retention and FHB severity than the Rht-B1b allele. Fusarium graminearum establishes infection sites predominantly inside the floral cavity and retained anthers potentially support colonization and initial hyphal growth, leading to a higher disease level in genotypes with a higher proportion of retained anthers. This is the first report demonstrating that differences in disease severity associated with Rht-D1b and Rht-B1b can be partly explained by their different effect on the extent of anther retention.

scholarly journals Functional Characterization of Calcineurin-Responsive Transcription Factors Fg01341 and Fg01350 in Fusarium graminearum

2020 ◽  
Vol 11 ◽  
Author(s):  
Xiangxiang Zhang ◽  
Shulin Cao ◽  
Wei Li ◽  
Haiyan Sun ◽  
Yuanyu Deng ◽  
...  
Keyword(s):  
Fusarium Graminearum ◽  
Type Strain ◽  
Deletion Mutant ◽  
Wild Type Strain ◽  
Functional Characterization ◽  
Hyphal Growth ◽  
Basic Medium ◽  
Dependent Manner ◽  
Wild Type ◽  
Sexual Production

Ca2 +/calmodulin-dependent phosphatase calcineurin is one of the important regulators of intracellular calcium homeostasis and has been investigated extensively in Saccharomyces cerevisiae. However, only a few reports have explored the function of the Crz1 homolog in filamentous fungi, especially in Fusarium graminearum. In this study, we identified Fg01341 as a potential ortholog of yeast Crz1. Fg01341 could interact with calcineurin and initiate nuclear transport in a calcineurin-dependent manner. The ΔFg01341 mutant exhibited normal hyphal growth on basic medium and conidia formation, but sexual reproduction was partially blocked. Pathogenicity assays showed that the virulence of the ΔFg01341 mutant in flowering wheat heads and corn silks dramatically decreased and was thus consistent with the reduction in deoxynivalenol production. Unexpectedly, the sensitivity to osmotic stress of the deletion mutant and that of the wild-type strain did not present any differences. The deletion mutant showed higher sensitivity to tebuconazole than the wild-type strain. Results also showed that the transcription factor Fg01350 might be the calcineurin target and was independent of Crz1. Furthermore, ΔFg01350 showed defects in hyphal growth, sexual production, virulence, and deoxynivalenol production. Collectively, the results indicate that these two proteins functionally redundant and that the calcineurin–Crz1-independent pathway is particularly important in F. graminearum.

scholarly journals Detoxification and Excretion of Trichothecenes in Transgenic Arabidopsisthaliana Expressing Fusarium graminearum Trichothecene 3-O-acetyltransferase

Toxins ◽  
2021 ◽  
Vol 13 (5) ◽  
pp. 320
Author(s):  
Guixia Hao ◽  
Susan McCormick ◽  
Helene Tiley ◽  
Thomas Usgaard
Keyword(s):  
Fusarium Graminearum ◽  
Transgenic Arabidopsis ◽  
Plant Protection ◽  
Plant Cells ◽  
Wild Type ◽  
Head Blight ◽  
Protection Mechanism ◽  
The Media ◽  
Self Protection

Fusarium graminearum, the causal agent of Fusarium head blight (FHB), produces trichothecenes including deoxynivalenol (DON), nivalenol (NIV), and 3,7,15-trihydroxy-12,13-epoxytrichothec-9-ene (NX-3). These toxins contaminate grains and cause profound health problems in humans and animals. To explore exploiting a fungal self-protection mechanism in plants, we examined the ability of F. graminearum trichothecene 3-O-acetyltransferase (FgTri101) to detoxify several key trichothecenes produced by F. graminearum: DON, 15-ADON, NX-3, and NIV. FgTri101 was cloned from F. graminearum and expressed in Arabidopsis plants. We compared the phytotoxic effects of purified DON, NIV, and NX-3 on the root growth of transgenic Arabidopsis expressing FgTri101. Compared to wild type and GUS controls, FgTri101 transgenic Arabidopsis plants displayed significantly longer root length on media containing DON and NX-3. Furthermore, we confirmed that the FgTri101 transgenic plants acetylated DON to 3-ADON, 15-ADON to 3,15-diADON, and NX-3 to NX-2, but did not acetylate NIV. Approximately 90% of the converted toxins were excreted into the media. Our study indicates that transgenic Arabidopsis expressing FgTri101 can provide plant protection by detoxifying trichothecenes and excreting the acetylated toxins out of plant cells. Characterization of plant transporters involved in trichothecene efflux will provide novel targets to reduce FHB and mycotoxin contamination in economically important plant crops.

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 Biological Characteristics and Molecular Mechanisms of Fludioxonil Resistance in Fusarium graminearum in China

Plant Disease ◽  
2020 ◽  
Vol 104 (9) ◽  
pp. 2426-2433
Author(s):  
F. Zhou ◽  
D. X. Li ◽  
H. Y. Hu ◽  
Y. L. Song ◽  
Y. C. Fan ◽  
...  
Keyword(s):  
Fusarium Graminearum ◽  
Molecular Mechanisms ◽  
Target Genes ◽  
Stop Codon ◽  
Point Mutations ◽  
Premature Stop Codon ◽  
Cross Resistance ◽  
Alternative Mechanism ◽  
Head Blight ◽  
Resistant Mutants

Fusarium graminearum is the primary causal agent of Fusarium head blight (FHB) of wheat. The phenylpyrrole fungicide fludioxonil is not currently registered for the management of FHB in China. The current study assessed the fludioxonil sensitivity of a total of 53 F. graminearum isolates collected from the six most important wheat-growing provinces of China during 2018 and 2019. The baseline fludioxonil sensitivity distribution indicated that all of the isolates were sensitive, exhibiting a unimodal cure with a mean effective concentration for 50% inhibition value of 0.13 ± 0.12 μg/ml (standard deviation). Five fludioxonil-resistant mutants were subsequently induced by exposure to fludioxonil under laboratory conditions. Ten successive rounds of subculture in the absence of the selection pressure indicated that the mutation was stably inherited. However, the fludioxonil-resistant mutants were found to have reduced pathogenicity, higher glycerol accumulation, and higher osmotic sensitivity than the parental wild-type isolates, indicating that there was a fitness cost associated with fludioxonil resistance. In addition, the study also found a positive cross resistance between fludioxonil, procymidone, and iprodione, but not with other fungicides such as boscalid, carbendazim, tebuconazole, and fluazinam. Sequence analysis of four candidate target genes (FgOs1, FgOs2, FgOs4, and FgOs5) revealed that the HBXT2R mutant contained two point mutations that resulted in amino acid changes at K223T and K415R in its FgOs1 protein, and one point mutation at residue 520 of its FgOs5 protein that resulted in a premature stop codon. Similarly, the three other mutants contained point mutations that resulted in changes at the K192R, K293R, and K411R residues of the FgOs5 protein but none in the FgOs2 and FgOs4 genes. However, it is important to point out that the FgOs2 and FgOs4 expression of all the fludioxonil-resistant mutants was significantly (P < 0.05) downregulated compared with the sensitive isolates (except for the SQ1-2 isolate). It was also found that one of the resistant mutants did not have changes in any of the sequenced target genes, indicating that an alternative mechanism could also lead to fludioxonil resistance.

scholarly journals Adaptation of Fusarium graminearum to Tebuconazole Yielded Descendants Diverging for Levels of Fitness, Fungicide Resistance, Virulence, and Mycotoxin Production

2010 ◽  
Vol 100 (5) ◽  
pp. 444-453 ◽  
Author(s):  
Rayko Becher ◽  
Ursula Hettwer ◽  
Petr Karlovsky ◽  
Holger B. Deising ◽  
Stefan G. R. Wirsel
Keyword(s):  
Fusarium Graminearum ◽  
Fungicide Resistance ◽  
Quantitative Polymerase Chain Reaction ◽  
Sublethal Dose ◽  
Cross Resistance ◽  
Azole Resistance ◽  
Head Blight ◽  
Widespread Application ◽  
Mycotoxin Production ◽  
Azole Fungicides

Azole fungicides play a prominent role for reliable plant disease management. However, quantitative azole resistance has been shown to develop in fungal pathogens, including Fusarium graminearum, the causal agent of Fusarium head blight (FHB). Due to widespread application of azole fungicides, resistance may accumulate to higher degrees in fungal field populations over time. Although azole fungicides are prominent components in FHB control, little effort has been made to investigate azole resistance in F. graminearum. We allowed F. graminearum strain NRRL 13383 to adapt to an azole fungicide in vitro, applying a strongly growth-reducing but sublethal dose of tebuconazole. Two morphologically distinguishable azole-resistant phenotypes were recovered that differed with regard to levels of fitness, fungicide resistance, virulence, and mycotoxin production. Isolates of the adapted “phenotype 1” exhibited azole-specific cross-resistance, whereas “phenotype 2” isolates displayed the phenomenon of multidrug resistance because the sensitivity to amine fungicides was also affected. Assessment of individual infected spikelets for mycotoxin contents by high-performance liquid chromatography mass spectrometry and for Fusarium DNA by quantitative polymerase chain reaction indicated that some of the adapted isolates produced significantly higher levels of nivalenol per fungal biomass than the NRRL 13383 strain.

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