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The plant rapid alkalinization factor (RALF) peptides function as key regulators in cell growth and immune responses through the receptor kinase FERONIA (FER). In this study, we report that the transcription factor FgPacC binds directly to the promoter of FgRALF gene, which encodes a functional homologue of the plant RALF peptides from the wheat head blight fungus Fusarium graminearum (FgRALF). More importantly, FgPacC promotes fungal infection via host immune suppression by activating the expression of FgRALF. The FgRALF peptide also exhibited typical activities of plant RALF functions, such as inducing plant alkalinization and inhibiting cell growth, including wheat (Triticum aestivum), tomato (Solanum lycopersicum) and Arabidopsis thaliana. We further identified the wheat receptor kinase FERONIA (TaFER), which is capable of restoring the defects of the A. thaliana FER mutant. In addition, we found that FgRALF peptide binds to the extracellular malectin-like domain (ECD) of TaFER (TaFER ECD ) to suppress the PAMP-triggered immunity (PTI) and cell growth. Overexpression of TaFER ECD in A. thaliana confers plant resistance to F. graminearum and protects from FgRALF-induced cell growth inhibition. Collectively, our results demonstrate that the fungal pathogen-secreted RALF mimic suppresses host immunity and inhibits cell growth via plant FER receptor. This establishes a novel pathway for the development of disease-resistant crops in the future without compromising their yield potential., (© 2024 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.)

Indoleamine 2,3-dioxygenase (Ido) is a tryptophan-degrading enzyme that is widely distributed across species. Ido catalyzes the first step of tryptophan (TRP) degradation and drives the de novo synthesis of nicotinamide adenine dinucleotide (NAD + ) coenzymes via the kynurenine (KYN) pathway. The budding yeast Saccharomyces cerevisiae possesses a single IDO gene (BNA2) that is responsible for NAD + synthesis, whereas a number of fungal species contain multiple IDO genes. However, the biological roles of IDO paralogs in plant pathogens remain unclear. In the current study, we identified three FgIDOs from the wheat head blight fungus Fusarium graminearum. FgIDOA/B/C expression was significantly induced upon TRP treatment. Targeted disruption of FgIDOA and/or FgIDOB caused different levels of NAD + auxotrophy, thus resulting in pleotropic phenotypic defects. Loss of FgIDOA resulted in abnormal conidial morphology, reduced mycelial growth, decreased virulence in wheat heads and reduced deoxynivalenol accumulation. Exogenous addition of KYN or various intermediates involved in the KYN pathway rescued auxotrophy of the mutants. Metabolomics analysis revealed shifts toward alternative TRP degradation pathways to melatonin and indole derivatives in mutants lacking FgIDOB. Upregulation of partner genes in auxotrophic mutants and the capacity to rescue the auxotroph by overexpressing a partner gene indicated functional complementation among FgIDOA/B/C. Taken together, the results of this study provide insights into differential roles in paralogous FgIDOs and how fungal TRP catabolism modulates fungal development and virulence., Competing Interests: Declaration of Competing Interest The authors have declared that no competing interests exist., (Copyright © 2023. Published by Elsevier GmbH.)

The Fusarium graminearum species complex (FGSC) can produce various mycotoxins and is a major concern for food quantity and quality worldwide. In this study, we determined the effects of water activity (a w ), temperature, incubation time and their interactions on mycotoxin accumulation and the expression levels of biosynthetic genes in FGSC strains from maize samples in China. The highest deoxynivalenol (DON), 3-acetyldeoxynivalenol(3ADON) and 15-acetyldeoxynivalenol (15ADON) levels of the F. boothii and F. graminearum strains were observed at 0.98 a w /30 °C or 0.99 a w /25 °C. F. asiaticum and F. meridionale reached maximum nivalenol (NIV) and 4-acetylnivalenol (4ANIV) contents at 0.99 a w and 30 °C. With the extension of the incubation time, the concentrations of DON and NIV gradually increased, while those of their derivatives decreased. F. boothii, F. meridionale and one F. asiaticum strain had the highest zearalenone (ZEN) values at 0.95 a w and 25 °C, while the optimum conditions for the other F. asiaticum strain and F. graminearum were 0.99 a w and 30 °C. Four genes associated with trichothecene and zearalenone synthesis were significantly induced under higher water stress in the early stage of production. The results indicated independence of mycotoxin production and gene expression, as maximum amounts of these toxic metabolites were observed at higher a w in most cases. This study provides useful information for the monitoring and prevention of such toxins entering the maize production chain., (© 2023. The Author(s), under exclusive licence to Microbiological Society of Korea.)

Fusarium ear rot (FER) is a serious fungal disease occurring the late growth stage of maize. FER not only reduces the yield of maize but also causes mycotoxin contamination, which affects the quality of maize and threatens human and animal health. Fusarium verticillioides is the predominant causative pathogen of FER worldwide. At present, there is no registered fungicide for use against maize FER in China. The novel isopropyl alcohol-triazole fungicide mefentrifluconazole (MFZ) has been shown to be effective against several Fusarium spp., but little is known about its specific activity against F. verticillioides. MFZ exhibited strong antifungal activities against 50 strains of F. verticillioides collected from the major maize-growing areas in China. MFZ inhibited mycelial growth, conidium production, germination and germ tube elongation of F. verticillioides. MFZ treatment significantly reduced fumonisin production and the expression levels of fumonisin biosynthetic genes. Genome-wide transcriptional profiling of F. verticillioides in response to MFZ indicated that the expression of genes involved in ergosterol biosynthesis, including fungicide target genes (cyp51 genes), was significantly downregulated by MFZ. MFZ treatment resulted in reduced ergosterol production and increased glycerol and malonaldehyde production as well as relative conductivity in F. verticillioides. A 2-year field experiment showed a significant reduction in FER severity in maize after spraying with MFZ at the tasseling stage. This study evaluated the potential of MFZ to control FER in maize and provides insights into its antifungal activities and mechanism of action against F. verticillioides., Competing Interests: Declaration of Competing Interest The authors declare that there is no conflict of interest., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Fusarium fujikuroi species complex (FFSC) strains are a major concern for food quantity and quality due to their strong ability to synthesize mycotoxins. The effects of interacting conditions of water activity, temperature, and incubation time on the growth rate, toxin production, and expression level of biosynthetic genes were examined. High temperature and water availability increased fungal growth. Higher water activity was in favor of toxin accumulation. The maximum amounts of fusaric acid (FA) and fumonisin B1 (FB1) were usually observed at 20-25 °C. F. andiyazi could produce a higher content of moniliformin (MON) in the cool environment than F. fujikuroi . The expression profile of biosynthetic genes under environmental conditions varied wildly; it was suggested that these genes might be expressed in a strain-dependent manner. FB1 concentration was positively related to the expression of FUM1 , while a similar correlation of FUB8 and FUB12 with FA production could be observed in F. andiyazi , F. fujikuroi , and F. subglutinans . This study provides useful information in the monitoring and prevention of such toxins entering the maize production chain.

An antibiotic produced by Paenibacillus polymyxa 7F1 was studied. The 7F1 strain was isolated from the rhizosphere of a wheat field. Response surface methodology was used to optimize the physicochemical parameters. The strain showed broad-spectrum activity against several plant pathogens. Identification of the strain was realized based on 16s rRNA gene and gyrB gene sequencing. The antibiotic was optimized by one-factor-at-a-time (OFAT) and response surface methodology (RSM) approaches. The suitable antibiotic production conditions were optimized using the one-factor-at-a-time method. The individual and interaction effects of three independent variables: culture temperature, initial pH, and culture time, were optimized by Box-Behnken design. The 16SrRNA gene sequence (1239 nucleotides) and gyrB gene (1111 nucleotides) were determined for strain 7F1 and shared the highest identities to those of Paenibacillus polymyxa . The results showed the optimal fermentation conditions for antibiotics produced by Paenibacillus polymyxa 7F1 were a culture temperature of 38 °C, initial pH of 8.0, and culture time of 8 h. The antibiotics produced by Paenibacillus polymyxa 7F1 include lipopeptides such as iturin A and surfactin. The results provide a theoretical basis for the development of bacteriostatic biological agents and the control of mycotoxins.

Fusarium crown rot of wheat is a serious fungal disease that occurs worldwide. The disease has been emerging in the major wheat-growing areas in China since 2010. Fusarium pseudogramineaum is the predominant causative pathogen of crown rot of wheat in China. The 14α-demethylation inhibitor (DMI) fungicide metconazole has been shown to be effective against Fusarium spp., but little is known about its specific activity against F. pseudogramineaum. Metconazole exhibited strong antifungal activities against all thirty-nine F. pseudogramineaum strains collected from the major wheat-growing areas in China. Metconazole inhibited mycelial growth and conidial germ tube elongation of F. pseudograminearum. Metconazole treatment significantly reduced the production of major toxins and the expression levels of toxin biosynthesis genes. Genome-wide transcriptional profiling of F. pseudograminearum in response to metconazole indicated that the expression of genes involved in ergosterol biosynthesis, including fungicide target genes (cyp51 genes), was significantly induced by metconazole. Nine ATP-binding cassette (ABC) transporter-encoding genes were significantly expressed in response to metconazole treatment. Reduced ergosterol production and antioxidant enzyme activities were observed after metconazole treatment. Greenhouse experiments indicated a significant reduction in crown rot occurrence in wheat after seed treatment with metconazole. This study evaluated the potential of metconazole to manage wheat crown rot and provides information to understand its antifungal activities and mechanism of action against F. pseudograminearum., Competing Interests: Declaration of Competing Interest The authors declare no competing interests., (Copyright © 2022. Published by Elsevier Inc.)

Autophagy is the main intracellular degradation system by which cytoplasmic materials are transported to and degraded in the vacuole/lysosome of eukaryotic cells, and it also controls cellular differentiation and virulence in a variety of filamentous fungi. However, the contribution of the autophagic pathway to fungal development and pathogenicity in the important maize pathogen and mycotoxigenic fungus Fusarium verticillioides is still unknown. In this study, we characterized two autophagy-related proteins, FvAtg4 and FvAtg8. The F. verticillioides deletion mutants Δ FvAtg4 and Δ FvAtg8 were impaired in autophagosome formation, aerial hyphal formation, sexual growth, lipid turnover, pigmentation and fungal virulence. Interestingly, Δ FvAtg4 and Δ FvAtg8 were defective in fumonisin B1 (FB1) synthesis, which may have resulted from decreased intracellular levels of alanine in the mutants. Our results indicate that FvAtg4 and FvAtg8 contribute to F. verticillioides pathogenicity by regulating the autophagic pathway to control lipid turnover, fumonisin biosynthesis, and pigmentation during its infectious cycle.

Aims: Cereals contaminated with type B trichothecene nivalenol (NIV) and its acetylated derivative 4-acetyl-nivalenol (4-AcNIV) are a global mycotoxicological problem threatening the health of humans and livestock. Toxicological studies, quantitative determinations and screening for biodegrading micro-organisms require massive amounts of pure toxins. However, the low yield from fungal cultures and high prices of NIV and 4-AcNIV limit research progress in these areas. This work aimed to select Fusarium asiaticum mutant strains with enhanced production of NIV and 4-AcNIV., Methods and Results: A total of 62 NIV-producing F. asiaticum strains were isolated and compared regarding their ability to produce NIV. Strain RR108 had the highest yield of NIV among 62 field isolates surveyed and was then genetically modified for higher production. Targeted deletion of the FaFlbA gene, encoding a regulator of G protein signalling protein, resulted in a significant increase in NIV and 4-AcNIV production in the FaFlbA deletion mutant ΔFaFlbA. The expression of three TRI genes involved in the trichothecene biosynthetic pathway was upregulated in ΔFaFlbA. ΔFaFlbA produced the highest amount of NIV and 4-AcNIV when cultured in brown long-grain rice for 21 days, and the yields were 2.07 and 2.84 g kg -1 , respectively. The mutant showed reduced fitness, including reduced conidiation, loss of perithecial development and decreased virulence on wheat heads, which makes it biologically safe for large-scale preparation and purification of NIV and 4-AcNIV., Conclusions: The F. asiaticum mutant strain ΔFaFlbA presented improved production of NIV and 4-AcNIV with reduced fitness and virulence in plants., Significance and Impact of the Study: Targeted deletion of the FaFlbA gene resulted in increased NIV and 4-AcNIV production. Our results provide a practical approach using genetic modification for large-scale mycotoxin production., (© 2021 The Society for Applied Microbiology.)

Members of the Fusarium graminearum species complex (FGSC) cause extensive yield losses in cereal production worldwide, and food safety concerns due to the accumulation of Fusarium toxins in infected grains. Among these pathogens, F. meridionale is responsible for Fusarium head blight of wheat and rice, ear and stalk rot of maize, and pod blight of soybean. Here, we present an improved genome assembly of F. meridionale strain SR5 isolated from rice in China based on PacBio long-read sequencing and Illumina short-read sequencing technology. The assembled genome of SR5 has a total size of 36.82 Mb, an N 50 scaffold length of 7.82 Mb, nine scaffolds, and encodes 12,409 predicted genes. These high-quality data expand FGSC genomic resources and provide a valuable resource for better understanding their genetic diversity and the molecular basis of pathogenesis, which will facilitate the development of an effective control strategy.[Formula: see text] Copyright © 2021 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Fusarium graminearum and Fusarium verticillioides are fungal pathogens that cause diseases in cereal crops, such as Fusarium head blight (FHB), seedling blight, and stalk rot. They also produce a variety of mycotoxins that reduce crop yields and threaten human and animal health. Several strategies for controlling these diseases have been developed. However, due to a lack of resistant cultivars and the hazards of chemical fungicides, efforts are now focused on the biocontrol of plant diseases, which is a more sustainable and environmentally friendly approach. In the present study, the lipopeptide mycosubtilin purified from Bacillus subtilis ATCC6633 significantly suppressed the growth of F. graminearum PH-1 and F. verticillioides 7600 in vitro. Mycosubtilin caused the destruction and deformation of plasma membranes and cell walls in F. graminearum hyphae. Additionally, mycosubtilin inhibited conidial spore formation and germination of both fungi in a dose-dependent manner. In planta experiments demonstrated the ability of mycosubtilin to control the adverse effects caused by F. graminearum and F. verticillioides on wheat heads and maize kernels , respectively. Mycosubtilin significantly decreased the production of deoxynivalenol (DON) and B-series fumonisins (FB 1 , FB 2 and FB 3 ) in infected grains, with inhibition rates of 48.92, 48.48, 52.42, and 59.44%, respectively. The qRT-PCR analysis showed that mycosubtilin significantly downregulated genes involved in mycotoxin biosynthesis. In conclusion, mycosubtilin produced by B . subtilis ATCC6633 was shown to have potential as a biological agent to control plant diseases and Fusarium toxin contamination caused by F. graminearum and F. verticillioides.

Fusarium incarnatum - equiseti species complex (FIESC) strains are generally considered moderately virulent to many agricultural crops and produce a variety of mycotoxins, which represent a serious threat to food safety and public health. The occurrence of the FIESC strain in agricultural crops has been reported in various climatic regions, but detailed information on the species composition and toxigenic ability is rare in China. In this study, phylogenetic analyses were performed with combined sequences of EF - 1a and RPB2 of 186 Fusarium isolates obtained from rice ( Oryza sativa ) and soybean ( Glycine max ). Twelve species were identified and 156 of the isolates were resolved within the Incarnatum clade of the FIESC species. Host influenced the population composition: rice isolates belonged to 12 species, among which FIESC 16, 18, and 24 strains were predominant; whereas five species were found among soybean isolates and FIESC 1, 16, and 18 strains dominated. Forty-three isolates were arbitrarily selected and analyzed for their Tri gene sequences and mycotoxigenic potential. Phylogenetic results based on the combined Tri 5, Tri 7, and Tri13 sequences were coincident with those from housekeeping markers. Type-A and -B trichothecenes were the main metabolites. Diacetoxyscirpenol was detected in all strains at varying concentrations. Nivalenol, 4-acetyl nivalenol, 3-acetyl deoxynivalenol, and neosolaniol were produced in members of the FIESC 1, 3, 7, 8, 15, 16, 17, and 18 strains. Our findings contribute valuable phylogenetic and toxigenic information necessary for the risk evaluation of mycotoxins in agricultural products.

Species belonging to the Fusarium fujikuroi species complex (FFSC) are of vital importance and are a major concern for food quantity and quality worldwide, as they not only cause serious diseases in crops but also produce various mycotoxins. To characterize the population structure and evaluate the risk of poisonous secondary metabolites, a total of 237 candidate strains were isolated from rice, maize, and soybean samples in Jiangsu Province, China. Species identification of the individual strain was accomplished by sequencing the translation elongation factor 1α gene ( TEF-1α ) and the fumonisin (FB) synthetic gene ( FUM1 ). The distribution of Fusarium species among the different crops was observed. The maize seeds were dominated by F. proliferatum (teleomorph, Gibberella intermedia ) and F. verticillioides (teleomorph, G. moniliformis ), whereas F. fujikuroi (teleomorph, G. fujikuroi ) was the most frequently isolated species from rice and soybean samples. In addition, phylogenetic analyses of these strains were performed, and the results suggested clear groups showing no obvious relationship with the origin source. FFSC species pathogenicity and toxigenicity were studied. All of the species reduced the rice seed germination rate, with no significant differences. F. fujikuroi showed two distinct patterns of influencing the length of rice seedlings, which were correlated with FBs and gibberellic acid synthesis. FBs were mainly produced by F. verticillioides and F. proliferatum. F. proliferatum and F. fujikuroi also produced moniliformin and beauvericin. The toxigenicity of F. andiyazi (teleomorph, G. andiyazi ) was extremely low. Further analysis indicated that the sequence variations in TEF-1α and the differences in the expression levels of the toxin synthesis genes were associated with the diversity of secondary metabolites in F. fujikuroi strains. These findings provide insight into the population-level characterization of the FFSC and might be helpful in the development of strategies for the management of diseases and mycotoxins.

Fusarium head blight (FHB), caused by the fungal pathogen Fusarium graminearum, is a destructive and widespread wheat disease. Chemical fungicides are becoming less effective at reducing the disease severity of FHB, and there is a need to find a more effective, low-cost natural product. A by-product of the pyrolysis of wheat straw is a condensate known as wheat straw vinegar, which was hypothesized to be an effective F. graminearum inhibitor in wheat. The organic and mineral compositions of wheat straw vinegar were analyzed. The results of GC-MS indicated that the major organic compounds in wheat straw vinegar are phenolics and acetic acid. The main inorganic elements in the liquid were K, Ca, S and Mg. A bio-test of wheat straw vinegar showed strong antifungal activity on F. graminearum growth and production of deoxynivalenol (DON) with an EC 50 (concentration for 50% of maximal effect) value of 3.1 μl ml -1 . Field tests showed that the application of wheat straw vinegar diluted 200-fold significantly decreased the wheat FHB infection rate and DON content by 66% and 69%, respectively. The control efficacy of wheat straw vinegar at a dilution of 200-fold was similar to that of typical chemical fungicide applications. The use of wheat straw vinegar may increase farmers' income by reducing the net fungicide costs. Therefore, wheat straw vinegar has high potential as a natural fungicide for the control of FHB and can reduce the dependence on synthetic fungicides., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Dihydroxyacid dehydratase (DHAD), encoded by ILV3, catalyses the third step in the biosynthetic pathway of branched-chain amino acids (BCAAs), which include isoleucine (Ile), leucine (Leu), and valine (Val). Enzymes involved in BCAA biosynthesis exist in bacteria, plants, and fungi but not in mammals and are therefore attractive targets for antimicrobial or herbicide development. In this study, three paralogous ILV3 genes (FgILV3A, FgILV3B, and FgILV3C) were identified in the genome of Fusarium graminearum, the causal agent of Fusarium head blight (FHB). Deletion of FgILV3A alone or combined with FgILV3B or FgILV3C indicated an important role for FgILV3A in BCAA biosynthesis. FgILV3A deletion mutants lost the ability to grow on medium lacking amino acids. Exogenous supplementation of 1 mM Ile and Val rescued the auxotrophy of ΔFgIlv3A, though 5 mM was required to recover the growth defects in ΔFgIlv3AB and ΔFgIlv3AC strains, indicating that FgIlv3b and FgIlv3c exhibit redundant but accessory roles with FgIlv3a in BCAA biosynthesis. The auxotrophy of ΔFgIlv3A resulted in pleiotropic defects in aerial hyphal growth, in conidial formation and germination, and in aurofusarin accumulation. In addition, the mutants showed reduced virulence and deoxynivalenol production. Overall, our study demonstrates that FgIlv3a is crucial for BCAA biosynthesis in F. graminearum and a candidate fungicide target for FHB management.

Wheat Fusarium head blight (FHB), caused by Fusarium species, is a widespread and destructive fungal disease. In addition to the substantial yield and revenue losses, diseased grains are often contaminated with Fusarium mycotoxins, making them unsuitable for human consumption or use as animal feed. As a vital food and feed ingredient in China, the quality and safety of wheat and its products have gained growing attention from consumers, producers, scientists, and policymakers. This review supplies detailed data about the occurrence of Fusarium toxins and related intoxications from the 1980s to the present. Despite the serious situation of toxin contamination in wheat, the concentration of toxins in flour is usually lower than that in raw materials, and food-poisoning incidents have been considerably reduced. Much work has been conducted on every phase of toxin production and wheat circulation by scientific researchers. Regulations for maximum contamination limits have been established in recent years and play a substantial role in ensuring the stability of the national economy and people's livelihoods., Competing Interests: The authors declare no conflict of interest.

3-isopropylmalate dehydrogenase (IPMD) encoded by LEU2 is a key enzyme in leucine (Leu) biosynthetic pathway. Analysis of the genome sequence of Fusarium graminearum revealed two paralogous LEU2 genes (designated as FgLEU2A and FgLEU2B) in this fungus and the deduced amino acid sequences of FgLeu2A and FgLeu2B share 45% identity. Targeted disruption of individual FgLEU2A/B gene in F. graminearum assigned a more crucial role of FgLeu2A in Leu biosynthesis as disruption of FgLEU2A resulted in mutant (ΔFgLeu2A-10) that was Leu-auxotrophic and could not grow in minimal medium limited for amino acids, whereas FgLEU2B deletion mutant ΔFgLeu2B-2 was morphologically indistinguishable from the wild type strain PH-1. The growth defects of ΔFgLeu2A-10 could be overcome by exogenous addition of Leu at 0.25 mM. Double deletion of FgLEU2A and FgLEU2B (ΔFgLeu2AB-8) caused a more severe Leu-auxotrophic phenotype as the concentration of Leu exogenously added to medium to rescue the growth defect of ΔFgLeu2AB-8 should be raised to 1.25 mM, indicating a less important but nonnegligible role of FgLeu2B in Leu biosynthesis. Disturb of Leu biosynthesis caused by FgLEU2A deletion leads to slower growth rate, reduced aerial hyphal formation and red pigmentation on PDA plates and completely blocked conidial production and germination. All of the defects above could be overcome by Leu addition or complementation of the full-length FgLEU2A gene. ΔFgLeu2A-10 also showed significantly increased sensitivity to osmotic and oxidative stresses. Pathogenicity assay results showed that virulence of mutants lacking FgLEU2A were dramatically impaired on wheat heads and non-host cherry tomatoes. Additionally, a low level of deoxynivalenol (DON) production of ΔFgLeu2A-10 and ΔFgLeu2AB-8 in wheat kernels was also detected. Taken together, results of this study indicated a crucial role of FgLeu2A and a less important role of FgLeu2B in Leu biosynthesis and fungal infection-related morphogenesis in F. graminearum and FgLeu2A may serve as a potential target for novel antifungal development., Competing Interests: The authors have declared that no competing interests exist.

Background: The Fusarium graminearum species complex infects several cereals and causes the reduction of grain yield and quality. Many factors influence the extent of Fusarium infection and mycotoxin levels. Such factors include crop rotation. In the present study, we explored the effect of rice or maize as former crops on mycotoxin accumulation in wheat grains., Results: More than 97% of samples were contaminated with deoxynivalenol (DON). DON concentrations in wheat grains from rice and maize rotation fields were 884.37 and 235.78 µg kg(-1) . Zearalenone (ZEN) was detected in 45% of samples which were mainly collected from maize-wheat rotation systems. Fusarium strains were isolated and more F. graminearum sensu stricto (s. str.) isolates were cultured from wheat samples obtained from maize rotation fields. DON levels produced by Fusarium isolates from rice rotation fields were higher than those of samples from maize rotation fields., Conclusions: Rice-wheat rotation favours DON accumulation, while more ZEN contamination may occur in maize-wheat rotation models. Appropriate crop rotation may help to reduce toxin levels in wheat grains. © 2016 Society of Chemical Industry., (© 2016 Society of Chemical Industry.)

The present study was performed to identify prevailing Fusarium species and the environmental factors affecting their frequencies and the contamination of grain with major mycotoxins in Jiangsu province. The precipitation levels were 184.2mm, 156.4mm, and 245.8mm in the years 2013-2015, respectively, and the temperature fluctuated by an average of 10.6±7.2°C in 2013, 10.9±7.2°C in 2014, and 10.6±6.3°C in 2015. Co-occurrence of deoxynivalenol (DON), 3-acetyldeoxynivalenol (3ADON), and 15-acetyldeoxynivalenol (15ADON) were observed in wheat. The average concentrations of DON were 879.3±1127.8, 627.8±640.5, and 1628.6±2,168.0μg/kg in 2013-2015, respectively. The average concentrations of 3ADON were 43.5±59.0, 71.2±102.5, and 33.5±111.9μg/kg in 2013-2015, respectively. We found that the average concentration of DON in wheat was positively correlated with precipitation (r=0.998, p<0.01), and 3ADON was negatively correlated with precipitation (r=-0.887, p<0.05). However, there was no correlation between precipitation and 15ADON or nivalenol (NIV). The differences in temperature were not as significant as the differences in rainfall amount over a short time period. Therefore, there were no correlations between temperature and the concentrations of trichothecenes, excluding 3ADON (r=0.996, p<0.01). Our data indicated that Fusarium asiaticum is the primary pathogenic fungus prevalent in the Fusarium head blight disease nursery. The trichothecene chemotype composition differed between Fusarium graminearum sensu stricto (s. str.) and F. asiaticum isolates. The 3ADON chemotype was found only among strains of F. asiaticum. The NIV chemotype was not observed among strains of F. graminearum, while the 15ADON chemotype represented 100% of the F. graminearum strains collected. The results of this study indicated no correlations between environmental conditions and the species or genetic chemotype composition of pathogens in Jiangsu province in 2013-2015., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Fusarium head blight (FHB) resistant line Soru#1 was hybridized with the German cultivar Naxos to generate 131 recombinant inbred lines for QTL mapping. The population was phenotyped for FHB and associated traits in spray inoculated experiments in El Batán (Mexico), spawn inoculated experiments in Ås (Norway) and point inoculated experiments in Nanjing (China), with two field trials at each location. Genotyping was performed with the Illumina iSelect 90K SNP wheat chip, along with a few SSR and STS markers. A major QTL for FHB after spray and spawn inoculation was detected on 2DLc, explaining 15-22% of the phenotypic variation in different experiments. This QTL remained significant after correction for days to heading (DH) and plant height (PH), while another QTL for FHB detected at the Vrn-A1 locus on 5AL almost disappeared after correction for DH and PH. Minor QTL were detected on chromosomes 2AS, 2DL, 4AL, 4DS and 5DL. In point inoculated experiments, QTL on 2DS, 3AS, 4AL and 5AL were identified in single environments. The mechanism of resistance of Soru#1 to FHB was mainly of Type I for resistance to initial infection, conditioned by the major QTL on 2DLc and minor ones that often coincided with QTL for DH, PH and anther extrusion (AE). This indicates that phenological and morphological traits and flowering biology play important roles in resistance/escape of FHB. SNPs tightly linked to resistance QTL, particularly 2DLc, could be utilized in breeding programs to facilitate the transfer and selection of those QTL.

In this study, we characterized FgIlv2 and FgIlv6, the catalytic and regulatory subunits of acetohydroxyacid synthase (AHAS) from the important wheat head scab fungus Fusarium graminearum. AHAS catalyzes the first common step in the parallel pathways toward branched-chain amino acids (BCAAs: isoleucine, leucine, valine) and is the inhibitory target of several commercialized herbicides. Both FgILV2 and FgILV6 deletion mutants were BCAA-auxotrophic and showed reduced aerial hyphal growth and red pigmentation when cultured on PDA plates. Conidial formation was completely blocked in the FgILV2 deletion mutant ΔFgIlv2-4 and significantly reduced in the FgILV6 deletion mutant ΔFgIlv6-12. The auxotrophs of ΔFgIlv2-4 and ΔFgIlv6-12 could be restored by exogenous addition of BCAAs but relied on the designated nitrogen source the medium contained. Deletion of FgILV2 or FgILV6 also leads to hypersensitivity to various cellular stresses and reduced deoxynivalenol production. ΔFgIlv2-4 lost virulence completely on flowering wheat heads, whereas ΔFgIlv6-12 could cause scab symptoms in the inoculated spikelet but lost its aggressiveness. Taken together, our study implies the potential value of antifungals targeting both FgIlv2 and FgIlv6 in F. graminearum.

A total of 122 wheat varieties obtained from the Nordic Genetic Resource Center were infected artificially with an aggressive Fusariumasiaticum strain in a field experiment. We calculated the severity of Fusarium head blight (FHB) and determined the deoxynivalenol (DON) content of wheat grain, straw and glumes. We found DON contamination levels to be highest in the glumes, intermediate in the straw, and lowest in the grain in most samples. The DON contamination levels did not increase consistently with increased FHB incidence. The DON levels in the wheat varieties with high FHB resistance were not necessarily low, and those in the wheat varieties with high FHB sensitivity were not necessarily high. We selected 50 wheat genotypes with reduced DON content for future research. This study will be helpful in breeding new wheat varieties with low levels of DON accumulation.

In this study we characterized FgIlv1, a homologue of the Saccharomyces cerevisiae threonine dehydratase (TD) from the important Fusarium head blight fungus Fusarium graminearum. TD catalyzes the first step in the biosynthesis pathway of isoleucine (Ile) for conversion of threonine (Thr) to 2-ketobutyrate (2-KB). The FgILV1 deletion mutant ΔFgIlv1-3 was unable to grow on minimal medium or fructose gelatin agar which lacked Ile. Exogenous supplementation of Ile or 2-KB but not Thr rescued the mycelial growth defect of ΔFgIlv1-3, indicating the involvement of FgIlv1 in the conversion of Thr to 2-KB in Ile biosynthesis. Additionally, exogenous supplementation of Methionine (Met) could also rescue the mycelial growth defect of ΔFgIlv1-3, indicating a crosstalk between Ile biosynthesis and Met catabolism in F. graminearum. Deletion of FgILV1 also caused defects in conidial formation and germination. In addition, ΔFgIlv1-3 displayed decreased virulence on wheat heads and a low level of deoxynivalenol (DON) production in wheat kernels. Taken together, results of this study indicate that FgIlv1 is an essential component in Ile biosynthesis and is required for various cellular processes including mycelial and conidial morphogenesis, DON biosynthesis, and full virulence in F. graminearum. Our data indicate the potential of targeting Ile biosynthesis for anti-FHB management.

A three-year (2010-2012) survey was conducted to assess the prevalence and concentrations of deoxynivalenol (DON) and zearalenone (ZEN) in wheat from several regions of Jiangsu province, China, which are heavily impacted by Fusarium head blight. A total of 180 wheat samples were obtained from the infected fields that spread 21 counties. DON and ZEN levels were determined by liquid chromatography-mass spectrometry (LC-MS/MS). DON was found in 74.4% of samples at levels ranging from 14.52 to 41157.13 μg/kg (mean 488.02 μg/kg), while ZEN was found in 12.8% of samples at levels ranging from 10.13 to 3048.88 μg/kg (mean 73.04 μg/kg). In years and regions of higher rainfall, DON and ZEN levels were higher in samples. These results are necessary to take a vigilant attitude to prevent human intake of trichothecenes and protect human's health from the risk of exposure to these toxins., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Members of the Fusarium graminearum species complex (FGSC) are important pathogens on wheat, maize, barley, and rice in China. Harvested grains are often contaminated by mycotoxins, such as the trichothecene nivalenol (NIV) and deoxynivalenol (DON) and the estrogenic mycotoxin zearalenone (ZEN), which is a big threat to humans and animals. In this study, 97 isolates were collected from maize, wheat, and rice in Jiangsu and Anhui provinces in 2013 and characterized by species- and chemotype-specific PCR. F. graminearum sensu stricto (s. str.) was predominant on maize, while most of the isolates collected from rice and wheat were identified as F. asiaticum. Fusarium isolates from three hosts varied in trichothecene chemotypes. The 3-acetyldeoxynivalenol (3ADON) chemotype predominated on wheat and rice population, while 15ADON was prevailing in the remaining isolates. Sequence analysis of the translation elongation factor 1α and trichodiene synthase indicated the accuracy of the above conclusion. Additionally, phylogenetic analysis suggested four groups with strong correlation with species, chemotype, and host. These isolates were also evaluated for their sensitivity to carbendazim and mycotoxins production. The maize population was less sensitive than the other two. The DON levels were similar in three populations, while those isolates on maize produced more ZEN. More DON was produced in carbendazim resistant strains than sensitive ones, but it seemed that carbendazim resistance had no effect on ZEN production in wheat culture.

In this study, we characterized FgIlv5, a homologue of the Saccharomyces cerevisiae keto-acid reductoisomerase (KARI) from the important wheat head scab fungus Fusarium graminearum. KARI is a key enzyme in the branched-chain amino acid (BCAA, including leucine, isoleucine and valine) biosynthetic pathway that exists in a variety of organisms from bacteria to fungi and higher plants, but not in mammals. The FgILV5 deletion mutant ΔFgIlv5-4 failed to grow when the culture medium was nutritionally limited for BCAAs. When grown on potato-dextrose agar plates, ΔFgIlv5-4 exhibited a significant decrease in aerial hyphae formation and red pigmentation. Conidia formation was also blocked in ΔFgIlv5-4. Exogenous addition of 1 mM isoleucine and valine was able to rescue the defects of mycelial growth and conidial morphogenesis. Cellular stress assays showed that ΔFgIlv5-4 was more sensitive to osmotic and oxidative stresses than the wild-type strain. In addition, virulence of ΔFgIlv5-4 was dramatically reduced on wheat heads, and a low level of deoxynivalenol production was detected in ΔFgIlv5-4 in wheat kernels. The results of this study indicate that FgIlv5 is involved in valine and isoleucine biosynthesis and is required for full virulence in F. graminearum.

Fusarium head blight (FHB) is a destructive wheat disease of global importance. Resistance breeding depends heavily on the Fhb1 gene. The CIMMYT line Shanghai-3/Catbird (SHA3/CBRD) is a promising source without this gene. A recombinant inbred line (RIL) population from the cross of SHA3/CBRD with the German spring wheat cv. Naxos was evaluated for FHB resistance and related traits in field trials using spray and spawn inoculation in Norway and point inoculation in China. After spray and spawn inoculation, FHB severities were negatively correlated with both anther extrusion (AE) and plant height (PH). The QTL analysis showed that the Rht-B1b dwarfing allele co-localized with a QTL for low AE and increased susceptibility after spawn and spray inoculation. In general, SHA3/CBRD contributed most of the favorable alleles for resistance to severity after spray and spawn inoculation, while Naxos contributed more favorable alleles for reduction in FDK and DON content and resistance to severity after point inoculation. SHA3/CBRD contributed a major resistance QTL close to the centromere on 2DLc affecting FHB severity and DON after all inoculation methods. This QTL was also associated with AE and PH, with high AE and tall alleles contributed by SHA3/CBRD. Several QTL for AE and PH were detected, and low AE or reduced PH was always associated with increased susceptibility after spawn and spray inoculation. Most of the other minor FHB resistance QTL from SHA3/CBRD were associated with AE or PH, while the QTL from Naxos were mostly not. After point inoculation, no other QTL for FHB traits was associated with AE or PH, except the 2DLc QTL which was common across all inoculation methods. Marker-assisted selection based on the 2DLc QTL from SHA3/CBRD combined with phenotypic selection for AE is recommended for resistance breeding based on this valuable source of resistance.

Fusarium head blight (FHB or scab) caused by Fusarium species is a destructive disease in wheat and barley worldwide. The objectives of our study were to identify quantitative trait loci (QTLs) for resistance to FHB spread (Type II resistance) and to quantify the magnitude of their effects in a novel highly resistant wheat germplasm, CJ 9306. A set of 152 F(7) recombinant inbred lines (RILs) derived from a cross Veery/CJ 9306 and two parents were evaluated for FHB resistance by single-floret inoculation in three greenhouse experiments in 2002 and 2004. Percentage (PSS) and number (NSS) of scabby spikelets at 25 days post-inoculation were analyzed. In total 682 simple sequence repeat (SSR) markers were screened for polymorphism between the two parents, and a genetic linkage map was constructed with 208 polymorphic markers. Ten QTLs associated with FHB resistance were detected, five from CJ 9306 and five from Veery. The major QTL on 3BS (QFhs.ndsu-3BS) was validated in CJ 9306, exhibiting greatest additive effects and explained 30.7% of phenotypic variation for PSS on the overall average of three experiments. Another major QTL on 2DL (QFhs.nau-2DL) from CJ 9306 explained 9.9-28.4% of phenotypic variation, with a significant QTL x environment interaction. QFhs.nau-1AS and QFhs.nau-7BS showed lower additive effects and explained lower variance (4.5-9.5%). A QTL on 5AS, decreasing PSS by 10.3% on average, was validated by simple marker analysis and joint trait/experiment IM/CIM analysis despite insignificance for single-experiment IM and CIM analyses. Likewise, QFhs.nau-2BL and QFhs.nau-1BC from Veery could reduce PSS by 13.2 and 11.4%, respectively. The effects of other three minor QTLs from Veery were significant for one experiment and combined analysis. Comparisons of two- and three-locus combinations suggested that the effects of FHB resistance QTLs/genes could be accumulated, and the resistance could be feasibly enhanced by selection of favorable alleles for multiple loci. Four two-locus combinations and two three-locus combinations were suggested as the preferential choices in practical marker-assisted selection program.

Fusarium head blight (FHB or scab) caused by Fusarium species is a destructive disease in wheat, not only causing dramatic decrease of grain yield and quality, but also leading to serious mycotoxin contamination in the infected grains. This study was conducted to identify and quantify quantitative trait loci (QTLs) contributing to resistance to deoxynivalenol (DON) accumulation as well as to grain yield loss in a population of 152 F(7) recombinant inbred lines (RILs) derived from the cross Veery/CJ 9306. DON content in scabby grains and relative decreases of yield components were analyzed. Two new QTLs (QFhs.nau-2DL and QFhs.nau-1AS) for resistance to DON accumulation caused by FHB in wheat were detected, and QTLs QFhs.ndsu-3BS and QFhs.nau-5AS were also validated in CJ 9306, based on a constructed genetic linkage map. On the average of three experiments, major QTLs QFhs.ndsu-3BS and QFhs.nau-2DL explained up to 23 and 20% of phenotypic variation, respectively. QFhs.nau-1AS and QFhs.nau-5AS separately explained 4-6% of phenotypic variation. The differences among years/experiments were significant for all the four QTLs. However, the QTL x environment interaction was significant only for QFhs.nau-2DL, but not for the others. The results suggest that simple sequence repeat (SSR) markers Xgwm533b associated with QFhs.ndsu-3BS, and Xgwm539 associated with QFhs.nau-2DL could be used in marker-assisted selection to enhance resistance to DON accumulation. QFhs.ndsu-3BS + QFhs.nau-2DL and QFhs.nau-2DL + QFhs.nau-5AS would be the optimum choices for two-locus combinations. QFhs.ndsu-3BS was also validated in CJ 9306 for resistance to grain yield loss, explaining 8-15% of phenotypic variation. No QTLs for resistance to DON accumulation or grain yield loss independent of Type II resistance were found. By comparison, however, either of QFhs.nau-2DL or QFhs.nau-5AS alone and their combination were more contributive to resistance to DON accumulation than to Type II resistance.

The main component of creosote obtained from dry wood distillation—guaiacol—is a natural antioxidant that has been widely used in pharmaceutical and food preservation applications. However, the antifungal mechanism of guaiacol against phytopathogens remains unclear. In this study, we found that guaiacol exerts inhibitory effects against mycelial growth, conidial formation and germination, and deoxynivalenol (DON) biosynthesis in Fusarium graminearum in a dose-dependent manner. The median effective concentration (EC50) value of guaiacol for the standard F. graminearum strain PH-1 was 1.838 mM. Guaiacol strongly inhibited conidial production and germination. The antifungal effects of guaiacol may be attributed to its capability to cause damage to the cell membrane by disrupting Ca2+ transport channels. In addition, the decreased malondialdehyde (MDA) levels and catalase (CAT), peroxidase (POD), and superoxide dismutase (SOD) activity by guaiacol treatment indicate that guaiacol displays activity against DON production by modulating the oxidative response in F. graminearum. Taken together, this study revealed the potentials of antioxidant in inhibiting mycotoxins in F. graminearum. [ABSTRACT FROM AUTHOR]

All organisms must secure essential trace elements (e.g., Cu) for survival and reproduction. However, excess trace element accumulation in cells is highly toxic. The maintenance of copper (Cu) homeostasis has been extensively studied in mammals, bacteria, and yeast but not in plant pathogens. In this study, we investigated the molecular mechanisms of copper tolerance in Fusarium graminearum , the important wheat head scab fungus. RNA-seq revealed induced expression of the P-type ATPase transporter FgCrpA and metallothionein (MT) FgCrdA after excess Cu treatment. Deletion of FgCrpA but not FgCrdA resulted in reduced tolerance to Cu toxicity. The "Cu fist" transcription factor FgAceA was involved in Cu detoxification through activation of FgCrpA. △FgAceA was more sensitive to copper toxicity than △FgCrpA and overexpression of FgCrpA restored copper tolerance in △FgAceA. FgAceA negatively regulated aurofusarin production and its biosynthetic gene expression. △FgCrpA and △FgAceA were reduced in virulence in flowering wheat heads and synthesized decreased amounts of the mycotoxin deoxynivalenol when challenged with excess Cu. Taken together, these results suggest that mediation of Cu tolerance in F. graminearum mainly relies on the Cu efflux pump and that FgAceA governs Cu detoxification through activation of FgCrpA. [ABSTRACT FROM AUTHOR]

Fusarium head blight (FHB) isa destructive wheat (Triticum aestivum L.) disease of global importance. The widely used dwarfing allele Rht-Dlb has recently been shown to compromise FHB resistance. The objectives of this study were to investigate the impact of this dwarfing allele in a segregating population with major resistance quantitative trait loci (OIL) derived from 'Sumai 3' and Nobeokabozu, and to determine how many resistance OIL are needed to counteract its negative effect. Fusarium head blight resistance was evaluated in four field trials with spray inoculation and two field trials with point inoculation in a double-haploid (DH) population from a cross between the Swedish cv. Avie (susceptible spring type; wild-type allele Rht-Dla) and Line 685 (resistant winter type; semi-dwarf allele Rht-Dlb). The Rht-D1 locus explained up to 38% of the phenotypic variation and was the most important QTL for FHB severity under spray inoculation but did not show any~ effect after point inoculation. Fhbl on 3B5 was detected with both inoculation methods but was relatively more important after point inoculation. Another two OIL on 5A and 2BL were detected after spray inoculation and a QTL on 2D after point inoculation. Comparison of phenotypic effects of different allele combinations revealed that a combination of both Fhbl and the 5A QTL was required to counteract the increased susceptibility of Rht-Dlb. Although breeding of FHB resistant cultivars with this dwarfing allele is possible, it requires the pyramiding of several resistant QTL to achieve adequate levels of resistance. [ABSTRACT FROM AUTHOR]

Fusarium graminearum is the predominant pathogen related to Fusarium head blight (FHB) which causes mycotoxin contamination of grains, mainly involving zearalenone (ZEN) and deoxynivalenol. The present work aimed to isolate a bacterium capable of antagonizing various phytopathogenic fungi and degrading ZEN effectively. Through co-culture with Fusarium graminearum , the bacterial strain MQ01 was isolated from wheat rhizosphere soil for its antifungal activity and identified as Bacillus amyloliquefaciens according to its morphological, physiological and biochemical characteristics and phylogenetic trees based on sequences of the 16 S rRNA and gyr B genes. B. amyloliquefaciens MQ01 showed broad-spectrum antifungal activity against eight phytopathogens, namely F. oxysporum , Sclerotinia sclerotiorum , F. proliferatum , F. verticillioides , Colletotrichum gloeosporioides , Microdochium nivale , F. equiseti and F. semitectum. The antifungal substances were extracted, separated by native-PAGE and identified through MALDI-TOF-MS. Protein homology analysis showed that the two antagonistic components were chitin-binding protein and subtilisin, respectively. In addition, MQ01 showed a prominent degradation capacity of ZEN both in liquid medium (94.1%) and in practical samples such as corn (27.9%) and distiller's dried grains with solubles (35.3%). One of the degradation product was identified as 1-(3,5-dihydroxyphenyl)-6′-hydroxy-l'-undecen-l0′-one by TOF-MS/MS analysis. In conclusion, B. amyloliquefaciens MQ01 shows promising prospects for use as a biocontrol agent against FHB and for ZEN degradation under field conditions. • An antagonistic bacterial strain against Fusarium graminearum was isolated and identified as Bacillus amyloliquefaciens MQ01. • The inhibitory rates of MQ01 against eight phytopathogenic fungi ranged from 17.4% to 57.3%. • The antifungal substances were separated and identified as chitin-binding protein and subtilisin by MALDI-TOF-MS. • Bacillus amyloliquefaciens MQ01 was able to eliminate zearalenone to an extent of 27.9% in corn and 35.3% in DDGS. [ABSTRACT FROM AUTHOR]