Fungus Fusarium Graminearum Research Articles

MeJA inhibits fungal growth and DON toxin production by interfering with the cAMP-PKA signaling pathway in the wheat scab fungus Fusarium graminearum.

Deoxynivalenol (DON), a mycotoxin primarily produced by Fusarium species, is commonly found in cereal grains and poses risks to human and animal health, as well as global grain trade. This study demonstrates that methyl jasmonate (MeJA), a natural plant hormone, inhibits the growth and conidiation of Fusarium graminearum. Importantly, MeJA significantly reduces DON production by suppressing TRI gene expression and toxisome formation. To explore the molecular mechanism, we identified MeJA-tolerant mutants, including a transcription factor MRT1 and cAMP-PKA pathway-related genes (FgGPA1 and FgSNT1). MeJA treatment reduced PKA activity and intracellular cAMP levels in F. graminearum, suggesting it targets the cAMP-PKA pathway. Notably, the MeJA-resistant mutant FgGPA1R178H enhanced fungal growth, DON production, and cAMP levels in the presence of MeJA. Exogenous cAMP alleviated MeJA's inhibitory effects on DON production, further supporting this pathway's involvement. Interestingly, MeJA had no effect on all three MAP kinase pathways (Mgv1, Gpmk1, and FgHog1). Truncated and phospho-mimicking mutations in Mrt1 or FgSnt1 conferred MeJA resistance, suggesting they may act downstream of the cAMP-PKA pathway. In conclusion, MeJA presents a promising approach to control F. graminearum growth and DON production.IMPORTANCEDeoxynivalenol (DON) poses significant risks to both human and animal health and severely disrupts the global grain trade due to its prevalence as a common contaminant in wheat grains. With rising public concern over food safety, finding effective and sustainable methods to reduce DON contamination becomes increasingly urgent. In our study, we found that methyl jasmonate (MeJA), a natural plant hormone, can effectively inhibit the vegetative growth of F. graminearum and significantly reduce its DON toxin production. To explore the underlying molecular mechanism, we identified the mutations in MeJA-tolerant mutants and revealed that MeJA effectively exerts its antifungal activities by inhibiting the cAMP-PKA signaling pathway in F. graminearum. Our work provides a promising natural solution to reduce DON toxin contamination in cereal grains, enhancing food safety while decreasing the reliance on chemical fungicides and their associated environmental impact.

A Cedrane and a Fusarin from the Algicolous Fungus Fusarium graminearum 12Ⅱ2N and Response to Mycoparasitism.

The fungal genus Fusarium is a treasure-trove of structurally diverse secondary metabolites, contributed greatly by marine-derived strains. A new cedrane sesquiterpene, fusacedrol (1), and a new fusarin member, fusarin M (2), were isolated from F. graminearum 12Ⅱ2N that was isolated as an endophyte from the marine brown alga Sargassum sp. The planar structures of compounds 1 and 2 were incisively identified by analysis of spectroscopic data, inclusive of NMR and MS, and the relative configurations were ensured by NOESY correlations and a coupling constant. Quantum chemical calculations of specific optical rotations of compound 1 and cedrol solved the absolute configuration of compound 1. Compound 1 represents the first new cedrane derivative with a 5/5/6 tricyclic nucleus from marine-derived fungi, and this skeleton has also been reported rarely from other marine organisms. Compound 2 is an isomer of fusarin Y, and its production could be greatly reduced by coculture with Trichoderma flagellatum 12A1N of the same origin. The two isolates were assayed for inhibiting the bacterium Pseudoalteromonas citrea (bio-02684, a green-spot pathogen of Porphyra), but none of them were active unfortunately.

The Fusarium graminearum Effector Protease FgTPP1 Suppresses Immune Responses and Facilitates Fusarium Head Blight Disease.

Most plant pathogens secrete effector proteins to circumvent host immune responses, thereby promoting pathogen virulence. One such pathogen is the fungus Fusarium graminearum, which causes Fusarium Head Blight (FHB) disease on wheat and barley. Transcriptomic analyses revealed that F. graminearum expresses many candidate effector proteins during early phases of the infection process, some of which are annotated as proteases. However, the contributions of these proteases to virulence remains poorly defined. Here, we characterize a F. graminearum endopeptidase, FgTPP1 (FGSG_11164), that is highly upregulated during wheat spikelet infection and is secreted from fungal cells. To elucidate the potential role of FgTPP1 in F. graminearum virulence, we generated FgTPP1 deletion mutants (ΔFgtpp1) and performed FHB infection assays. Deletion of FgTPP1 reduced the virulence of F. graminearium as assessed by spikelet bleaching. Infection with wild-type F. graminearum induced full bleaching in about 50% of the spikes at 10-11 days post infection, while this fraction was reduced to between 18 and 27% when using ΔFgtpp1 mutants. Transient expression of green fluorescent protein (GFP)-tagged FgTPP1 revealed that FgTPP1 localizes, in part, to chloroplasts and attenuates chitin-mediated activation of mitogen-activated protein kinase (MAPK) signaling, reactive oxygen species production, and cell death induced by an autoactive disease resistance protein when expressed in planta. Notably, the FgTPP1 protein is conserved across the Ascomycota phylum, suggesting it may be a core effector among ascomycete plant pathogens. These properties make FgTPP1 an ideal candidate for decoy substrate engineering, with the goal of engineering resistance to FHB.

Innovative Infrared Spectroscopic Technologies for the Prediction of Deoxynivalenol in Wheat.

Mycotoxin contamination in cereals is a global food safety concern. One of the most common mycotoxins in grains is deoxynivalenol (DON), a secondary metabolite produced by the fungiFusarium graminearum and Fusarium culmorum. Exposure to DON can lead to adverse health effects in both humans and animals including vomiting, dizziness, and fever. Hence, the development of analytical technologies capable of predicting mycotoxin contamination levels in grains is crucial. In this study, we emphasize innovative infrared (IR) spectroscopic technologies for the prediction of DON in wheat along the food supply chain. The performance of an IR laser spectroscopic platform for on-site or laboratory confirmative analysis was evaluated. Furthermore, the performance of a handheld IR spectrometer for preliminary screening during transportation, storage, or harvesting was assessed. The accuracy of cross validation (AccCV) obtained with the laser spectrometer reached 92%, while the handheld IR spectrometer achieved 84.6%. Hence, both technologies prove significant potential for rapid mycotoxin detection.

Functional Analysis of Durum Wheat GASA1 Protein as a Biotechnological Alternative Against Plant Fungal Pathogens and a Positive Regulator of Biotic Stress Defense.

Plants are frequently challenged by a variety of microorganisms. To protect themselves against harmful invaders, they have evolved highly effective defense mechanisms, including the synthesis of numerous types of antimicrobial peptides (AMPs). Snakins are such compounds, encoded by the GASA (Gibberellic Acid-Stimulated Arabidopsis) gene family, and are involved in the response to biotic and abiotic stress. Here, we examined the function of the newly identified TdGASA1 gene and its encoded protein in Triticum durum subjected to different biotic stress-related simulants, such as mechanical injury, methyl jasmonate (MeJA), indole-3-acetic acid (IAA), salicylic acid (SA), hydrogen peroxide (H2O2), as well as infection with pathogenic fungi Fusarium graminearum and Aspergillus niger. We found that in durum wheat, TdGASA1 transcripts were markedly increased in response to these stress simulants. Isolated and purified TdGASA1 protein exhibited significant antifungal activity in the growth inhibition test conducted on eight species of pathogenic fungi on solid and liquid media. Transgenic Arabidopsis lines overexpressing TdGASA1 obtained in this study showed higher tolerance to detrimental effects of H2O2, MeJA, and ABA treatment. In addition, these lines exhibited resistance to Fusarium graminearum and Aspergillus niger, which was linked to a marked increase in antioxidant activity in the leaves under stress conditions. This resistance was correlated with the upregulation of pathogenesis-related genes (AtPDF1.2a, AtERF1, AtVSP2, AtMYC2, AtPR1, AtACS6, AtETR1, and AtLOX2) in the transgenic lines. Overall, our results indicate that TdGASA1 gene and its encoded protein respond ubiquitously to a range of biotic stimuli and seem to be crucial for the basal resistance of plants against pathogenic fungi. This gene could therefore be a valuable target for genetic engineering to enhance wheat resistance to biotic stress.

Synthesis and Antifungal Evaluation of Cinnamic Acid-Geraniol Hybrids as Potential Fungicides.

Cinnamic acid and geraniol are two well-known antifungal natural products and widely applied in food and cosmetics industries. To discover novel natural product-based fungicide candidates with more potent activity and good ecological compatibility for the management of plant diseases, a series of cinnamic acid-geraniol hybrids were prepared by means of molecular hybridization and their chemical structures were well confirmed by spectral analysis. The antifungal activities of the target compounds against three phytopathogenic fungi Fusarium graminearum, Gaeumannomyces graminis (Sacc.) Arx et Oliver var. tritici (Sacc.) Walker, and Valsa mali were evaluated. Among them, compounds 5 e and 5 f showed the remarkable antifungal activity against G. graminis with the EC50 values of 82.719 and 91.828 μg/mL, respectively; while compounds 5 f and 6 b exhibited the obvious antifungal activity against V. mali. It suggested that compound 5 f can be further optimized for the design of novel broad-spectrum fungicide molecules as the secondary lead compound. In addition, some interesting structure-antifungal activity relationships were obtained. This work will provide some reference and guidance for the further discovery of novel fungicide candidates based on cinnamic acid and geraniol.

A novel strategy for obtaining essential oil and flavonoids from Thymus vulgaris L. using polyethylene glycol-ultrasonic pretreatment followed by microwave hydrodistillation and extraction

As a medicinal plant with high economic value, the green and efficient separation of the main components (essential oil (EO) and flavonoids) from Thymus vulgaris L. can enhance its application prospects. The present study developed a polyethylene glycol-ultrasonic pretreatment followed by microwave hydrodistillation and extraction (PEG-U-MHDE) technique to distill EO and extract flavonoids simultaneously. The parameters involved in the PEG-ultrasonic pretreatment and microwave irradiation processes were investigated using single factor and BBD experiments, from which 25 % PEG200/400/600 (1:1:1, v/v/v) was found to have the highest extraction ability. Under the optimal conditions, the yield of EO, apigenin, and baicalein was 3.72 ± 0.15 μL/g, 0.76 ± 0.03 mg/g, and 0.96 ± 0.03 mg/g, respectively. The PEG-U-MHDE showed good stability, recovery, and repeatability for apigenin and baicalein. In addition, the separation efficiency of EO, apigenin, and baicalein obtained by PEG-U-MHDE was 0.19 μL/g/min, 37.85 μg/g/min, and 48.03 μg/g/min, respectively, and the EO contained 81.77 % oxygenated compounds, which were higher than other reference methods. The inhibition rate of EO (concentration of 0.5 μL/mL) against the agricultural pathogenic fungus (Fusarium graminearum) was 36.65 ± 1.28 %, which was speculated to be closely related to its main oxygenated compounds (thymol and carvacrol). In general, PEG-U-MHDE has the advantages of short time and high efficiency, and can realize the simultaneous separation of volatile EO and non-volatile components, which provides a theoretical basis for the comprehensive utilization of natural medicinal plants and crops.

A novel ormycovirus isolated from the plant-pathogenic fungus Fusarium graminearum.

In this study, we identified a novel mycovirus, Fusarium graminearum ormycovirus 1 (FgOV1), from the pathogenic fungus Fusarium graminearum. The virus has two RNA segments, RNA1 and RNA2, with lengths of 2,591 and 1,801 nucleotides, respectively, excluding the polyA tail. Each segment contains a single open reading frame (ORF). The ORF in RNA1 encodes an RNA-dependent RNA polymerase, while the ORF in RNA2 encodes a hypothetical protein. Phylogenetic analysis showed that FgOV1 belongs to the gammaormycovirus clade, whose members are related to betaormycoviruses. To our knowledge, this is the first report of an ormycovirus in Fusarium graminearum.

Oxaloacetate anaplerosis differently contributes to pathogenicity in plant pathogenic fungi Fusarium graminearum and F. oxysporum.

Anaplerosis refers to enzymatic reactions or pathways replenishing metabolic intermediates in the tricarboxylic acid (TCA) cycle. Pyruvate carboxylase (PYC) plays an important anaplerotic role by catalyzing pyruvate carboxylation, forming oxaloacetate. Although PYC orthologs are well conserved in prokaryotes and eukaryotes, their pathobiological functions in filamentous pathogenic fungi have yet to be fully understood. Here, we delve into the molecular functions of the ortholog gene PYC1 in Fusarium graminearum and F. oxysporum, prominent fungal plant pathogens with distinct pathosystems, demonstrating variations in carbon metabolism for pathogenesis. Surprisingly, the PYC1 deletion mutant of F. oxysporum exhibited pleiotropic defects in hyphal growth, conidiation, and virulence, unlike F. graminearum, where PYC1 deletion did not significantly impact virulence. To further explore the species-specific effects of PYC1 deletion on pathogenicity, we conducted comprehensive metabolic profiling. Despite shared metabolic changes, distinct reprogramming in central carbon and nitrogen metabolism was identified. Specifically, alpha-ketoglutarate, a key link between the TCA cycle and amino acid metabolism, showed significant down-regulation exclusively in the PYC1 deletion mutant of F. oxysporum. The metabolic response associated with pathogenicity was notably characterized by S-methyl-5-thioadenosine and S-adenosyl-L-methionine. This research sheds light on how PYC1-mediated anaplerosis affects fungal metabolism and reveals species-specific variations, exemplified in F. graminearum and F. oxysporum.

Metabolomic Analyses Reveal That IAA from Serratia marcescens Lkbn100 Promotes Plant Defense during Infection of Fusarium graminearum in Sorghum.

Global sorghum production has been significantly reduced due to the occurrence of sorghum root rot caused by the fungus Fusarium graminearum. The utilization of biocontrol microorganisms has emerged as an effective strategy. However, the underlying mechanisms remain unclear. Therefore, the aim of this study was to investigate the effectiveness of biocontrol bacteria in inducing sorghum resistance against sorghum root rot and explore the potential induced resistance mechanisms through metabolomics analysis. The results revealed that the biocontrol bacteria Lnkb100, identified as Serratia marcescens (GenBank: PP152264), significantly enhanced the resistance of sorghum against sorghum root rot and promoted its growth, leading to increased seed weight. Targeted metabolomics analysis demonstrated that the highest concentration of the hormone IAA (indole-3-acetic acid) was detected in the metabolites of Lnkb100. Treatment with IAA enhanced the activity of disease-related enzymes such as SOD, CAT, POD and PPO in sorghum, thereby improving its resistance against sorghum root rot. Further untargeted metabolomic analysis revealed that IAA treatment resulted in higher concentrations of metabolites involved in the resistance against F. graminearum, such as geniposidic acid, 5-L-Glutamyl-taurine, formononetin 7-O-glucoside-6″-O-malonate, as well as higher concentrations of the defense-related molecules volicitin and JA. Additionally, "secondary bile acid biosynthesis" and "glycerophospholipid metabolism" pathways were found to play significant roles in the defense response of sorghum against fungal infection. These findings provide a reliable theoretical basis for utilizing biocontrol microorganisms to control sorghum root rot.

Inhibition of fusarium graminearum growth and spore germination by a streptomyces amritsarensis strain capable of killing and growing on microcystis scum.

Developing energy-saving and ecofriendly strategies for treating harvested Microcystis biomass. Streptomyces amritsarensis HG-16 was first reported to effectively kill various morphotypes of natural Microcystis colonies at very high cell densities. Concurrently, HG-16 grown on lysed Microcystis maintained its antagonistic activity against plant pathogenic fungus Fusarium graminearum. It could completely inhibit spore germination and destroy mycelial structure of F. graminearum. Transcriptomic analysis revealed that HG-16 attacked F. graminearum in a comprehensive way: interfering with replication, transcription, and translation processes, inhibiting primary metabolisms, hindering energy production and simultaneously destroying stress-resistant systems of F. graminearum. The findings of this study provide a sustainable and economical option for resource reclamation from Microcystis biomass: utilizing Microcystis slurry to propagate HG-16, which can subsequently be employed as a biocontrol agent for managing F. graminearum.

Edeine B1 produced by Brevibacillus brevis reduces the virulence of a plant pathogenic fungus by inhibiting mitochondrial respiration.

Plant pathogenic fungi cause serious diseases, which result in the loss of crop yields and reduce the quality of crops worldwide. To counteract the escalating risks of chemical fungicides, interest in biological control agents to manage plant diseases has significantly increased. In this study, we comprehensively screened microbial culture filtrates using a yeast screening system to find microbes exhibiting respiratory inhibition activity. Consequently, we found a soil-borne microbe Brevibacillus brevis HK544 strain exhibiting a respiration inhibitory activity and identified edeine B1 (EB1) from the culture filtrate of HK544 as the active compound of the respiration inhibition activity. Furthermore, against a plant pathogenic fungus Fusarium graminearum, our results showed that EB1 has effects on multiple aspects of respiration with the downregulation of most of the mitochondrial-related genes based on transcriptome analysis, differential EB1-sensitivity from targeted mutagenesis, and the synergistic effects of EB1 with electron transport chain complex inhibitors. With the promising plant disease control efficacy of B. brevis HK544 producing EB1, our results suggest that B. brevis HK544 has potential as a biocontrol agent for Fusarium head blight.IMPORTANCEAs a necrotrophic fungus, Fusarium graminearum is a highly destructive pathogen causing severe diseases in cereal crops and mycotoxin contamination in grains. Although chemical control is considered the primary approach to control plant disease caused by F. graminearum, fungicide-resistant strains have been detected in the field after long-term continuous application of fungicides. Moreover, applying chemical fungicides that trigger mycotoxin biosynthesis is a great concern for many researchers. Biocontrol of Fusarium head blight (FHB) by biological control agents (BCAs) represents an alternative approach and could be used as part of the integrated management of FHB and mycotoxin production. The most extensive studies on bacterial BCAs-fungal communications in agroecosystems have focused on antibiosis. Although many BCAs in agricultural ecology have already been used for fungal disease control, the molecular mechanisms of antibiotics produced by BCAs remain to be elucidated. Here, we found a potential BCA (Brevibacillus brevis HK544) with a strong antifungal activity based on the respiration inhibition activity with its active compound edeine B1 (EB1). Furthermore, our results showed that EB1 secreted by HK544 suppresses the expression of the mitochondria-related genes of F. graminearum, subsequently suppressing fungal development and the virulence of F. graminearum. In addition, EB1 exhibited a synergism with complex I inhibitors such as rotenone and fenazaquin. Our work extends our understanding of how B. brevis HK544 exhibits antifungal activity and suggests that the B. brevis HK544 strain could be a valuable source for developing new crop protectants to control F. graminearum.

Con7 is a key transcription regulator for conidiogenesis in the plant pathogenic fungus Fusarium graminearum.

The ascomycete fungus Fusarium graminearum is the primary cause of head blight disease in wheat and barley, as well as ear and stalk rot in maize. Given the importance of conidia and ascospores in the disease cycle of F. graminearum, precise spatiotemporal regulation of these biological processes is crucial. In this study, we characterized the Magnaporthe oryzae Con7p ortholog and discovered that FgCon7 significantly influences various crucial aspects of fungal development and pathogenicity. Notably, overexpression of FgABAA partially restored developmental defects in the FgCON7 deletion mutant. ChIP-qPCR analysis confirmed a direct genetic link between FgABAA and FgCON7. Furthermore, our research revealed a clear correlation between FgCon7 and chitin accumulation and the expression of chitin synthase genes. These findings offer valuable insights into the genetic mechanisms regulating conidiation and the significance of mycelial differentiation in this plant pathogenic fungus.

FgPfn participates in vegetative growth, sexual reproduction, pathogenicity, and fungicides sensitivity via affecting both microtubules and actin in the filamentous fungus Fusarium graminearum.

Fusarium head blight (FHB), caused by Fusarium graminearum species complexes (FGSG), is an epidemic disease in wheat and poses a serious threat to wheat production and security worldwide. Profilins are a class of actin-binding proteins that participate in actin depolymerization. However, the roles of profilins in plant fungal pathogens remain largely unexplored. Here, we identified FgPfn, a homolog to profilins in F. graminearum, and the deletion of FgPfn resulted in severe defects in mycelial growth, conidia production, and pathogenicity, accompanied by marked disruptions in toxisomes formation and deoxynivalenol (DON) transport, while sexual development was aborted. Additionally, FgPfn interacted with Fgα1 and Fgβ2, the significant components of microtubules. The organization of microtubules in the ΔFgPfn was strongly inhibited under the treatment of 0.4 μg/mL carbendazim, a well-known group of tubulin interferers, resulting in increased sensitivity to carbendazim. Moreover, FgPfn interacted with both myosin-5 (FgMyo5) and actin (FgAct), the targets of the fungicide phenamacril, and these interactions were reduced after phenamacril treatment. The deletion of FgPfn disrupted the normal organization of FgMyo5 and FgAct cytoskeleton, weakened the interaction between FgMyo5 and FgAct, and resulting in increased sensitivity to phenamacril. The core region of the interaction between FgPfn and FgAct was investigated, revealing that the integrity of both proteins was necessary for their interaction. Furthermore, mutations in R72, R77, R86, G91, I101, A112, G113, and D124 caused the non-interaction between FgPfn and FgAct. The R86K, I101E, and D124E mutants in FgPfn resulted in severe defects in actin organization, development, and pathogenicity. Taken together, this study revealed the role of FgPfn-dependent cytoskeleton in development, DON production and transport, fungicides sensitivity in F. graminearum.

The ING protein Fng2 associated with RPD3 HDAC complex for the regulation of fungal development and pathogenesis in wheat head blight fungus

ING proteins display a high level of evolutionary conservation across various species, and play a crucial role in modulating histone acetylation levels, thus regulating various important biological processes in yeast and humans. Filamentous fungi possess distinct biological characteristics that differentiate them from yeasts and humans, and the specific roles of ING proteins in filamentous fungi remain largely unexplored. In this study, an ING protein, Fng2, orthologous to the yeast Pho23, has been identified in the wheat head blight fungus Fusarium graminearum. The deletion of the FNG2 gene resulted in defects in vegetative growth, conidiation, sexual reproduction, plant infection, and deoxynivalenol (DON) biosynthesis. Acting as a global regulator, Fng2 exerts negative control over histone H4 acetylation and governs the expression of over 4000 genes. Moreover, almost half of the differentially expressed genes in the fng3 mutant were found to be co-regulated by Fng2, emphasizing the functional association between these two ING proteins. Notably, the fng2 fng3 double mutant exhibits significantly increased H4 acetylation and severe defects in both fungal development and pathogenesis. Furthermore, Fng2 localizes within the nucleus and associates with the FgRpd3 histone deacetylase (HDAC) to modulate gene expression. Overall, Fng2's interaction with FgRpd3, along with its functional association with Fng3, underscores its crucial involvement in governing gene expression, thereby significantly influencing fungal growth, asexual and sexual development, pathogenicity, and secondary metabolism.

Genome-Wide Association Study of Fungicide Sensitivity in a Fusarium graminearum Population Collected from North Dakota.

Fusarium head blight is a destructive disease of small grains. The disease is predominantly caused by the haploid ascomycete fungus Fusarium graminearum in North America. To understand the genetics of quantitative traits for sensitivity to fungicides in this fungal pathogen, we conducted a genome-wide association study of sensitivity to two demethylation inhibition class fungicides, tebuconazole and prothioconazole, using an F. graminearum population of 183 isolates collected between 1981 and 2013 from North Dakota. Baseline sensitivity to tebuconazole and prothioconazole was established using 21 isolates collected between 1981 and 1994. Most fungal isolates were sensitive to both tebuconazole and prothioconazole; however, five isolates showed significantly reduced sensitivity to prothioconazole. The genome-wide association study identified one significant marker-trait association on chromosome 3 for tebuconazole resistance, whereas six significant marker-trait associations, one on chromosome 1, three on chromosome 2, and two on chromosome 4, were detected for prothioconazole resistance. Functional annotation of the marker-trait association for tebuconazole revealed a candidate gene encoding a basic helix-loop-helix domain-containing protein that reinforces sterol in the fungal membrane. Putative genes for prothioconazole resistance were also identified, which are involved in RNA interference, the detoxification by ubiquitin-proteasome pathway, and membrane integrity reinforcement. Considering the potential of the pathogen toward overcoming chemical control, continued monitoring of fungal sensitivities to commercially applied fungicides, especially those containing prothioconazole, is warranted to reduce risks of fungicide resistance in the pathogen populations.

Silence of five F. graminearum genes in wheat host confers resistance to Fusarium head blight

Fusarium head blight (FHB), mainly caused by fungus Fusarium graminearum (F. graminearum), is a devastating wheat disease worldwide, leading to reduced yield production and compromised grain quality due to contamination by mycotoxins, such as deoxynivalenol (DON). Manipulating the specific gene expression in microorganisms through RNA interference (RNAi) presents an opportunity for new-generation double-stranded RNA (dsRNA)-based formulations to combat a large number of plant diseases. Here, we applied both spray-induced gene silencing (SIGS) and host-induced gene silencing (HIGS) to target five virulence-related and DON-synthesized genes in F. graminearum, including protein kinase gene Gpmk1, zinc finger protein gene FgChy1, transcription factor FgSR, DON synthesis gene TRI5 and the cell-end marker protein gene FgTeaA, aiming to effectively control FHB in wheat. Direct spraying of individual or combined siRNAs (small interfering RNA) from the fungus showed reduced expression of target genes and suppressed pathogenic symptoms during F. graminearum infection in wheat leaves, with the combination of all five siRNAs demonstrating superior resistance. Furthermore, we generated transgenic wheat lines expressing chimeric RNAi cassettes targeting these five genes, and two independent lines exhibited strong resistance to FHB and Fusarium crown rot, and the reduced DON accumulation. Notably, the HIGS transgenic lines did not adversely impact plant growth and yield traits. Collectively, our findings support that SIGS and HIGS represent effective strategies targeting key pathogenic genes for bolstering disease resistance in crops.

Proteomic Analysis of Cell Wall Proteins with Various Linkages in Fusarium graminearum.

The fungal cell wall, primarily comprising a glucan-chitin matrix and cell wall proteins (CWPs), serves as a key mediator for fungal interactions with the environment and plays a pivotal role in virulence. In this study, we employed a comprehensive proteomics approach to analyze the CWPs in the plant pathogenic fungus Fusarium graminearum. Our methodology successfully extracted and identified 1373 CWPs, highlighting their complex linkages, including noncovalent bonds, disulfide bridges, alkali-sensitive linkages, and glycosylphosphatidylinositol (GPI) anchors. A significant subset of these proteins, enriched in Gene Ontology terms, suggest multifunctional roles of CWPs. Through the integration of transcriptomic and proteomic data, we observed differential expression patterns of CWPs across developmental stages. Specifically, we focused on two genes, Fca7 and Cpd1, which were upregulated in planta, and confirmed their localization predominantly outside the plasma membrane, primarily in the cell wall and periplasmic space. The disruption of FCA7 reduced virulence on wheat, aligning with previous findings and underscoring its significance. Overall, our findings offer a comprehensive proteomic profile of CWPs in F. graminearum, laying the groundwork for a deeper understanding of their roles in the development and interactions with host plants.

Ginger (Zingiber officinale)-Mediated Green Synthesis of Silver-Doped Tin Oxide Nanoparticles and Evaluation of Its Antimicrobial Activity.

Green synthesis of nanoparticles using plant extract is a novel development that has gained significant attention because of its low cost, nontoxicity, and environmental friendliness. In the present study, silver-doped stannic oxide (Ag-doped SnO2) nanoparticle was synthesized by an eco-friendly green synthesis method. The synthesized samples were characterized using scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), ultraviolet-visible spectroscopy (UV-vis), X-ray diffraction (XRD), and their antimicrobial activities were assessed against two bacteria Escherichia coli (E. coli) and Staphylococcus aureus (S. aurus) and two fungi Fusarium oxysporum (F. oxysporum) and Fusarium graminearum (F. graminearum) by using the disk diffusion method. Ag-doped SnO2 nanoparticles show a strong and broad absorption from UV-vis spectra when compared to pure SnO2 nanoparticles. FTIR spectral analysis revealed that the peak at 505.69 cm-1 was assigned to Sn-O and O-Sn-O stretching vibration of SnO2 nanoparticles. XRD analysis confirmed the formation of a tetragonal rutile structure with average particle size ranging from 10 to 17 nm. The antimicrobial result indicates that the Ag-doped SnO2 revealed significant antimicrobial activity against both bacterial and fungi strains with the zone of inhibition of 29 ± 0.54, 27 ± 0.05, 17 ± 0.05, and 15 ± 0.05 mm for S. aureus, E. coli, F. oxysporum , and F. graminearum, respectively. Thus, the studies suggested that Ag-doped SnO2 nanoparticles exhibit good activity against both Gram-negative and Gram-positive bacteria and fungi. This is because doping SnO2 nanoparticles with metallic elements such as Ag has been used to enhance their performance, confirming them as a good candidate for antimicrobial agents and the development of future therapeutic agents.

Identification and functional characterisation of a locus for target site integration in Fusariumgraminearum

BackgroundFusarium Head Blight (FHB) is a destructive floral disease of different cereal crops. The Ascomycete fungus Fusariumgraminearum (Fg) is one of the main causal agents of FHB in wheat and barley. The role(s) in virulence of Fg genes include genetic studies that involve the transformation of the fungus with different expression cassettes. We have observed in several studies where Fg genes functions were characterised that integration of expression cassettes occurred randomly. Random insertion of a cassette may disrupt gene expression and/or protein functions and hence the overall conclusion of the study. Target site integration (TSI) is an approach that consists of identifying a chromosomal region where the cassette can be inserted. The identification of a suitable locus for TSI in Fg would avert the potential risks of ectopic integration.ResultsHere, we identified a highly conserved intergenic region on chromosome 1 suitable for TSI. We named this intergenic region TSI locus 1. We developed an efficient cloning vector system based on the Golden Gate method to clone different expression cassettes for use in combination with TSI locus 1. We present evidence that integrations in the TSI locus 1 affects neither fungal virulence nor fungal growth under different stress conditions. Integrations at the TSI locus 1 resulted in the expression of different gene fusions. In addition, the activities of Fg native promoters were not altered by integration into the TSI locus 1. We have developed a bespoke bioinformatic pipeline to analyse the existence of ectopic integrations, cassette truncations and tandem insertions of the cassette that may occurred during the transformation process. Finally, we established a protocol to study protein secretion in wheat coleoptiles using confocal microscopy and the TSI locus 1.ConclusionThe TSI locus 1 can be used in Fg and potentially other cereal infecting Fusarium species for diverse studies including promoter activity analysis, protein secretion, protein localisation studies and gene complementation. The bespoke bioinformatic pipeline developed in this work together with PCR amplification of the insert could be an alternative to Southern blotting, the gold standard technique used to identify ectopic integrations, cassette truncations and tandem insertions in fungal transformation.