During the 1950s, the Gros Michel species of bananas were nearly wiped out by the incurable Fusarium Wilt, also known as Panama Disease. Originating in Southeast Asia, Fusarium Wilt is a banana pandemic that has been threatening the multi-billion-dollar banana industry worldwide. The disease is caused by a fungus that spreads rapidly throughout the soil and into the roots of banana plants. Currently, the only way to stop the spread of this disease is for farmers to manually inspect and remove infected plants as quickly as possible, whereas it is a time-consuming process. The main purpose of this study is to build a deep Convolutional Neural Network (CNN) using a transfer learning approach to rapidly identify fusarium wilt infections on banana crop leaves. We chose to use the ResNet50 architecture as the base CNN model for our transfer learning approach owing to its remarkable performance in image classification, which was demonstrated through its victory in the ImageNet competition. After its initial training and fine-tuning on a data set consisting of 300 healthy and diseased images, the CNN model achieved near-perfect accuracy of 0.99 and was fine-tuned to adapt the ResNet base model. ResNet50’s distinctive residual block structure could be the reason behind these results. To evaluate this CNN model, 500 test images, consisting of 250 diseased and healthy banana leaf images, were classified by the model. The deep CNN model was able to achieve an accuracy of 0.98 and an F-1 score of 0.98 by correctly identifying the class of 492 of the 500 images. These results show that this DCNN model outperforms existing models such as Sangeetha et al., 2023’s deep CNN model by at least 0.07 in accuracy and is a viable option for identifying Fusarium Wilt in banana crops.

Knowledge on the health of banana trees is critical for farmers to profit from banana cultivation. Fusarium wilt and banana blood diseases (BDB), two significant diseases infecting banana trees, are caused by Fusarium oxysporum and Ralstonia syzygii, respectively. They have successfully caused a decline in crop yield as they destroy the trees, starting sequentially from the pseudostem to the fruits. The entire distribution of BDB and Fusarium on a plantation can be understood using advanced geospatial information obtained from multispectral aerial photographs taken using an unmanned aerial vehicle (UAV), combined with the reliable data field of infected trees. Vegetation and soil indices derived from a multispectral aerial photograph, such as normalized difference vegetation index, modified chlorophyll absorption ratio index, normalized difference water index (NDWI) and soil pH, may have to be relied on to explain the precise location of these two diseases. In this study, a random forest algorithm was used to handle a large dataset consisting of two models: the banana diseases multispectral model and the banana diseases spectral model. The results show that the soil indices, soil pH and NDWI are the most important variables for predicting the spatial distribution of these two diseases. Simultaneously, the plantation area affected by BDB is more extensive than that affected by Fusarium, if the variation of planted banana cultivars is not considered.

Wilted hemp (Cannabis sativa L.) plants were observed in August 2019 in commercial fields around Osijek, Croatia. Plants and roots with disease symptoms were collected. The single-spored isolates produced septate cottony white to light pink aerial mycelium and purple undersurface on potato dextrose agar (PDA). Smooth and hyaline hyphae were branched and septate. Macroconidia were fusiform to sickle-shaped with foot-shaped basal cell, elongated apical cell and three to five septa. Sequencing of the internal transcribed spacer and the partial elongation factor 1-α gene identified species as Fusarium oxysporum. Artificial infection fulfills Koch’s postulates, producing plants which showed stunted growth and wilt symptoms similar to those observed in the commercial fields. Control seedlings remained symptomless and healthy. To the best of our knowledge, this is the first report of hemp Fusarium wilt causing F. oxysporum in Croatia. Considering that F. oxysporum has been reported in main field crops in Croatia, the presence of this pathogen could cause economically significant hemp production decreases, especially in humid and cold springs and susceptible varieties.

Fusarium wilt caused by Fusarium oxysporum f. sp. cubense (Foc) is a major disease affecting banana production around the globe. To study the movement of Foc in planta and investigate potential physical mechanisms of deterrence, the roots of a set of banana cultivars were challenged with a GFP-tagged subtropical race 4 strain of Foc. These plants included susceptible and resistant varieties including the plantain 'FHIA25' (AAB), dessert hybrids 'Lady Finger' (AAB), 'FHIA02' (AAAA), as well as the industry preferred Cavendish line, 'Williams' (AAA), and its somaclone derivative 'GCTCV119' (AAA). GFP localisation in planta confirmed that Foc can infect all four cultivars. Its movement through roots to the rhizome and to the aerial parts of the plants was evident. Tyloses formation in the root vasculature was also detected by GFP auto-fluorescence and further confirmed using scanning electron microscopy. While tyloses formation did not prevent colonisation and infection, their formation restricted the fungus within the xylem vessels, slowing its proliferation. This mechanism may be an important factor in providing plant host tolerance to Foc but it doesn't provide complete immunity. Fluorescently tagged Foc is a useful tool for understanding the infection process of the Fusarium wilt disease of banana and aiding in the integrated management of this important disease.

The effective management of Fusarium wilt of bananas (FW) depends on the knowledge of the disease dynamics in time and space. The objectives of this work were: To estimate disease intensity and impact, and to investigate the spatial and temporal dynamic of FW. Fields planted with Silk (n = 10), Pome (n = 17) or Cavendish (n = 3) banana subgroups were surveyed in Brazil, totaling 95 ha. In each field, all plants were visually assessed and diseased plants were georeferenced. The incidence of FW and the impact of the disease on yield on a regional scale were estimated. Spatial patterns were analyzed using quadrat- and distance-based methods. FW incidence ranged from 0.09 to 41.42%, being higher in Silk fields (median = 14.26%). Impacts of epidemics on yield ranged from 18.4 to 8,192.5 kg.ha-1.year-1, with a median of 935.2 kg.ha-1.year-1. The higher economic impact of the disease was observed on Silk cultivar with a median loss of US$ 910.5 ha-1.year-1. Overall, estimated losses increased on average by US$ 109.8 ha-1.year-1 at each 1% of incidence. Aggregation of FW was detected by all analytical methods in 13 fields (1 of Cavendish, 11 of Pome and 1 of Silk). In the other 17 fields, at least one analytical method did not reject the null hypothesis of randomness. One field (5 ha), composed of six plots, was selected for spatial and temporal studies during two years with bi-monthly assessments. A sigmoidal curve represented the FW progress and the Gompertz model best fitted disease progress. The level of aggregation varied over time, and evidence of secondary infection to neighboring and distant plants were detected. FW is a widespread problem in Brazil and yield losses can be of high magnitude. Epidemiology-based management strategies can now be better established.

Bananas, a staple food globally and a key agricultural commodity, face a severe threat from the fungus Fusarium oxysporum form specialis cubense (Foc), significantly impacting production. Genetic improvement to develop resistant cultivars stands out as a crucial strategy to mitigate this disease. This study focused on assessing and recommending enhanced diploid banana varieties for resistance against Foc subtropical race 4 (ST4) and Foc race 1 (R1). Twenty-five improved diploids developed by Embrapa, Brazil, underwent evaluation. Utilizing a scale for internal symptoms at 90 days post-inoculation, genotypes were categorized from highly resistant to highly susceptible based on the internal symptom index. The diploid M53 exhibited high resistance to R1 and resistance to Foc ST4, while only the diploid CNPMF 0534 demonstrated complete resistance to both R1 and ST4, with resistance to the latter likely associated with penetration. These findings provide valuable insights for banana and plantain breeding programs, offering selected diploids for crossbreeding with commercial cultivars to develop new, resistant genotypes against Foc.

Fusarium wilt (FW) caused by Fusarium oxysporum f. sp. Cucumerinum (Foc) of cucumber is a destructive soil-borne disease in cucumber (Cucumis sativus.L). There is limited knowledge on molecular mechanisms to FW resistance-mediated defense responses in cucumber. In this study, metabolome and transcriptome profiling were carried out in two FW, resistant (NR) and susceptible (NS), near isogenic lines (NILs) before and after Foc inoculation. The NILs showed consistent and stable FW resistances in multiple greenhouse and laboratory screening tests. Widely targeted metabolomic analysis identified differentially accumulated metabolites (DMAs) with significantly more accumulation in NR in response to Foc infection including many phenolic acid and flavonoid compounds from the flavonoid biosynthesis pathway. Transcriptome analysis identified differentially expressed genes (DEGs) between the NILs upon Foc inoculation including genes for secondary metabolite biosynthesis in or transcription factor genes regulating the flavonoid biosynthesis pathway. Joint analysis of the metabolomic and transcriptomic data identified DAMs and DEGs closely associated with biosynthesis of phenolic acid and flavonoid DAMs. The association of these compounds with NR-conferred FW resistance was exemplified by in vivo assays in which two phenolic acid compounds, bis (2-ethylhexyl) phthalate and diisooctyl phthalate, and the flavonoid compound gallocatechin 3-O-gallate were shown to have significant inhibitory effect on Foc growth. The antifungal effect of three compounds seems to be a novel finding. We conclude that phenolic acids and flavonoids play an important role in NR mediated FW resistance in cucumber.

Fusarium wilt diseases severely influence the growth and productivity of numerous crop plants. The consortium of antagonistic rhizospheric Bacillus strains and quercetin were evaluated imperatively as a possible remedy to effectively manage the Fusarium wilt disease of tomato plants. The selection of Bacillus strains was made based on in-vitro antagonistic bioassays against Fusarium oxysporum f.sp. lycoprsici (FOL). Whereas, quercetin was selected after screening a library of phytochemicals during in-silico molecular docking analysis using tomato LysM receptor Kinases “SILKY12” based on its dual role in symbiosis and plant defense responses. After the selection of test materials, pot trials were conducted where tomato plants were provided consortium of Bacillus strains as soil drenching and quercetin as a foliar spray in different concentrations. The combined application of consortium () and quercetin (1.0 mM) reduced the Fusarium wilt disease index up to 69% also resulting in increased plant growth attributes. Likewise, the imperative application of the Bacillus consortium and quercetin (1.0 mM) significantly increased total phenolic contents and activities of the enzymes of the phenylpropanoid pathway. Non-targeted metabolomics analysis was performed to investigate the perturbation in metabolites. FOL pathogen negatively affected a range of metabolites including carbohydrates, amino acids, phenylpropanoids, and organic acids. Thereinto, combined treatment of Bacillus consortium and quercetin (1.0 mM) ameliorated the production of different metabolites in tomato plants. These findings prove the imperative use of Bacillus consortium and quercetin as an effective and sustainable remedy to manage Fusarium wilt disease of tomato plants and to promote the growth of tomato plants under pathogen stress conditions.

Fusarium wilt caused by Fusarium oxysporum f. sp. cubense (Foc) is one of the most destructive diseases in banana farming worldwide. Knowledge of the factors of genetic diversity and virulence of the pathogen contributes to the development of resistant cultivars and in management strategies based on exclusion. In this study, phenotypic traits such as virulence and aggressiveness in a sample of 52 Foc isolates, were analyzed and their relationship to the presence of putative effectors of gene SIX (Secreted in Xylem) pathogenicity homologues, verified. According to the dissimilarity matrix, there were three isolates closest to the standard Foc race 1 strain. Isolates 229A and 218A, were selected according to their aggressiveness profile in 'Grand Naine' and 'Prata-Anã', respectively, I order to replace the standard isolate of race 1 in the resistance screening process carried out by the breeding program. Two homologues of the SIX8 gene, SIX8a and SIX8b, are present in isolates of Foc from Brazil and the SIX8b gene correlates with avirulence in the cultivar ‘Grand Naine’ (Cavendish). These results are important to support the banana genetic breeding program by identification of sources of resistance to Foc and contribute in the establishment of the function of SIX effector proteins.

Verticillium dahliae (Vd) and Fusarium oxysporum f. sp. lycopersici (Fol) are two major fungal pathogens that infect tomato plants, causing significant challenges in their control since both pathogens can persist in the soil for several years even in the absence of a host plant and no effective fungicides are available at present. This study investigated the efficacy of two biocontrol formulations, Clonotri and Strepse, against Vd and Fol, and their impact on tomato fruit quality and yield under greenhouse conditions. Pathogenicity experiment demonstrated that the Clonotri formulation, containing Trichoderma and Clonostachys spores, significantly reduced Fusarium wilt disease by 32% compared to the control group. However, in Vd pathogenicity experiment, the formulations did not exhibit disease reduction, although the treatment with Strepse, containing Streptomyces and Pseudomonas microorganisms, resulted in preserved total fruit number when compared to uninfected plants. Analysis of fruit quality attributes revealed no significant differences among the various interventions. Furthermore, Fol infection in the first fruit set significantly increased fruit firmness, while Vd infection resulted in elevated levels of total soluble solids in fruits. These findings demonstrate that the evaluated biocontrol formulations provide a degree of protection against fungal wilt pathogens in tomato plants and can increase yield in greenhouse conditions, while having minimal impact on overall fruit quality attributes.

In agriculture, soil-borne fungal pathogens, especially Fusarium oxysporum strains, are posing a serious threat to efforts to achieve global food security. In the search for safer agrochemicals, silica nanoparticles (SiO2NPs) have recently been proposed as a new tool to alleviate pathogen damage including Fusarium wilt. Hollow mesoporous silica nanoparticles (HMSNs), a unique class of SiO2NPs, have been widely accepted as a desired pesticide carrier. However, their role in enhancing the disease resistance and the specific mechanism remain unknown. In this study, three sizes of HMSNs (19, 96 and 406 nm as HMSNs–19, HMSNs–96 and HMSNs–406, respectively) were synthesized and characterized to determine their effects on seed germination, seedling growth, and Fusarium oxysporum f. sp. phaseolus (FOP) suppression in cowpea roots by foliar spray using phenotypes, fresh biomass and disease progression as indicators. The results revealed that three HMSNs exhibited no adverse impacts on seed germination and tended to improve plant growth. Also, they exert their FOP suppression with a size- and concentration-dependent manner. HMSNs–406 possessed the best control effect at a concentration of 1000 mg/L showing an upto 40.00% decline in the disease severity. The Si(OH)4 control was also effective on FOP suppression at a lower concentration of 100 mg/L, whereas its higher concentrations exhibited obviously adverse impacts on FOP control, seed germination and plant growth. Moreover, we conformed that HMSNs posed their Fusarium wilt suppression in cowpea plant by activating SA (salicylic acid)-dependent SAR (systemic acquired resistance) responses rather than directly suppressing FOP. A higher level of SA content and elevated expression of its maker genes of PR-1 and PR-5 in HMSNs–406 treated cowpea roots provided substantial evidences of this mode of action. Other resistance-related genes, as well as defense-responsive enzymes, were also involved in the HMSNs-activated SAR pathway. Overall, for the first time, our results extended a new role of HMSNs as a potent elicitor to serve as a versatile alternative for plant disease protection of low cost, highly efficiency and sustainability.

The promotion of plant growth and suppression of plant disease using beneficial microorganisms is considered an alternative to the application of chemical fertilizers or pesticides in the field. In this study, a coconut-scented antagonistic Trichoderma strain LZ42, previously isolated from Genoderma lucidum-cultivated soil, was investigated for biostimulatory and biocontrol functions in tomato seedlings. Morphological and phylogenetic analyses suggested that strain LZ42 is closely related to T. atroviride. Tomato plants showed increased aerial and root dry weights in greenhouse trials after treatment with T. atroviride LZ42 formulated in talc, indicating the biostimulatory function of this fungus. T. atroviride LZ42 effectively suppressed Fusarium wilt disease in tomato seedlings, with an 82.69% control efficiency, which is similar to that of fungicide treatment. The volatile organic compounds (VOCs) emitted by T. atroviride LZ42 were found to affect the primary root growth direction and promote the root growth of tomato seedlings in root Y-tube olfactometer assays. The fungal VOCs from T. atroviride LZ42 were observed to significantly inhibit F. oxysporum in a sandwiched Petri dish assay. SPME-GC-MS analysis revealed several VOCs emitted by T. atroviride LZ42; the dominant compound was tentatively identified as 6-pentyl-2H-pyran-2-one (6-PP). 6-PP exhibited a stronger ability to influence the direction of the primary roots of tomato seedlings but not the length of the primary roots. The inhibitory effect of 6-PP on F. oxysporum was the highest among the tested pure VOCs, showing a 50% effective concentration (EC50) of 5.76 μL mL-1 headspace. In conclusion, T. atroviride LZ42, which emits VOCs with multiple functions, is a promising agent for the biostimulation of vegetable plants and integrated management of Fusarium wilt disease.

Fusarium wilt, caused by Fusarium oxysporum f.sp. cubense (Foc), is recognized as one of the most devastating diseases affecting banana cultivation worldwide. In India, Foc extensively affects the Malbhog banana (AAB genomic group) production. In this study, we isolated 25 Foc isolates from wilt-affected Malbhog plantations of India. Pathogenicity test confirmed the identity of these isolates as Foc, the primary causative agent of wilt in banana. Morpho-cultural characterization of the Foc isolates showed large variation in colony morphological features, intensity, and pattern of pigmentation, chlamydospores, and conidial size. The molecular identification of these isolates using Race-1 and Race-4 specific primers established their identity as Race- 1 of Foc, with absence of TR- 4. For a more comprehensive understanding of genetic diversity of Foc isolates, we em-ployed ISSR molecular typing which revealed five major clusters. Ahigh96% diversity within Foc population indicated the presence of polymorphic loci in individuals of a given population evi-dent from results of Nei’s genetic diversity, Shannon information index, and polymorphism in-formation content values, apart from analysis of molecular variance (AMOVA). The current findings provide significant insights towards detection of Foc variants, and consequently the deployment of effective management practices to keep the possible epidemic development of disease under control along Malbhog banana growing belts of northeast India.

Banana is one of the most important fruits in the world due to its status as a major food source for more than 400 million people. Fusarium oxysporum f. sp. cubense tropical race 4 (Foc TR4) causes substantial losses of banana crops every year, and molecular host resistance mechanisms are currently unknown. We here performed a genome-wide analysis of the autophagy related protein 8 (ATG8) family in a wild banana species. The banana genome was found to contain 10 MaATG8 genes. Four MaATG8s formed a gene cluster in the distal part of chromosome 4. Phylogenetic analysis of ATG8 families in banana, Arabidopsis thaliana, Citrus, rice, and ginger revealed five major phylogenetic clades shared by all of these plant species, demonstrating evolutionary conservation of the MaATG8 families. The transcriptomic analysis of plants infected with Foc TR4 showed that almost all of the MaATG8 genes were more highly induced in resistant cultivars than in susceptible cultivars. Finally, MaATG8F was found to interact with MaATG4B in vitro (with yeast two-hybrid assays), and MaATG8F and MaATG4B all positively regulated banana resistance to Foc TR4. Our study provides novel insights into the structure, distribution, evolution, and expression of the MaATG8 family in bananas. Furthermore, the discovery of interactions between MaATG8F and MaATG4 could facilitate future researches of disease resistance genes for genetic improvement of bananas.

: Cnidium officinale Makino, a perennial crop in the Umbeliperae family, is harvested in the summer for medicinal purposes. The average temperature in Korea has recently increased, and consequently, the area used to cultivate C. officinale has been reduced, and plant disease and soil sickness have increased. In this study, rhizosphere bacteria were isolated from C. officinale, and their antagonistic activity evaluated against Fusarium solani, which is a major issue during repeated cultivation. Twenty-eight strains were isolated from the root and rhizosphere soil, of which seven showed antagonistic activity against F. solani and were further investigated. An in planta test found that the dead rate of shoots was significantly lower in the group inoculated with PT1 isolated from the root tissue and that the fresh and dry weights were also higher, when compared with the other groups. PT1 was thus considered to have antagonistic activity against F. solani. The 16S rRNA analysis identified the PT1 as Leclercia adecarboxylata, and downstream studies confirmed the production of enzymes related to antagonism, such as siderophore and N-acetyl-β-glucosaminidase. The phosphorous solubilizing ability and secretion of related enzymes were also confirmed. The results indicate that PT1 could be utilized as a PGPR to help control fusarium wilt.

Fusarium head blight (FHB) can cause contamination of cereal grain with mycotoxins. Triticale is also infected with FHB; however, it is more resistant than wheat to head infection. The aim of this study was to identify triticale lines that combine low head infection with low toxin contamination. Resistance to FHB of 15 winter triticale and three winter wheat lines was evaluated over a three-year experiment established in two locations. At the anthesis stage, heads were inoculated with Fusarium culmorum isolates. The FHB index was scored and the percentage of Fusarium-damaged kernels (FDKs) assessed. The grain was analysed for type B trichothecenes (deoxynivalenol and derivatives, nivalenol) and zearalenone content. The average FHB index was 10.7%. The proportion of FDK was 18.1% (weight) and 21.6% (number). An average content of deoxynivalenol for wheat amounted to 7.258 mg/kg and nivalenol to 5.267 mg/kg. In total, it was 12.788 m/kg of type B trichothecenes. The zearalenone content in the grain was 0.805 mg/kg. Relationships between FHB index, FDK and mycotoxin contents were statistically significant for triticale lines; however, they were stronger for FDK versus mycotoxins. Lines combing all types of FHB resistance were found, and two of them had resistance similar to that of wheat lines with the Fhb1 gene.

Fusarium dry rot is one of the major postharvest potato diseases of during storage after harvest, which not only results in quality degradation but also brings up huge economic losses. The dis-ease can be elicited by some species of Fusarium, the pathogenic fungi of Fusarium causing potato dry rot are considerably diverse in various countries and regions. Fusarium spp. infection is as-sociated with dry rot produce mycotoxins, which has phytotoxicity and mycotoxicoses on hu-mans and animals. Chemical synthetic fungicide is considered as the main control measure to Fusarium dry rot of potato, nevertheless, it is unfortunate that the persistent application inevitably results in the emergency of resistant strain and environmental contamination. A comprehensive disease control strategy includes potato cultivar selection, appropriate cultural practices (crop rotation, cultivate pattern, fertilization, and irrigation), harvesting process and postharvest treatment (harvesting, classification, packaging, wound healing), storage condition (environ-mental disinfection, temperature, humidity and gas composition) along with the application of fungicide pre-harvest or postharvest. Recently, emerging studies indicated that eco-friendly strategies include physical control, chemical methods (such as the application of generally rec-ognized as safe (GRAS) compounds or chemical (elicitors) and biological control have been in-troduced to combat Fusarium dry rot of potato.

The Fusarium fujikuroi species complex (FFSC) includes socioeconomically important pathogens that cause disease and/or mycotoxin contamination on numerous crops. Here, we used comparative genomics to elucidate processes underlying the ability of pine-associated and grass-associated FFSC species to colonize tissues of their respective plant hosts. We characterized the identity, possible functions, evolutionary origins, and chromosomal positions of the host-range-associated genes encoded by the two groups of fungi. The 72 and 47 genes identified as unique to the respective genome groups were potentially involved in diverse processes, ranging from transcription, regulation, and substrate transport, through to virulence/pathogenicity. Most emerged early during the evolution of Fusarium/FFSC and were subsequently retained only in some lineages, while some had origins outside Fusarium. Although differences in the densities of these genes were especially noticeable on the conditionally dispensable chromosome of F. temperatum (representing the grass-associates) and F. circinatum (representing the pine-associates), the host-range-associated genes tended to be located towards the subtelomeric regions of chromosomes. Taken together, these results demonstrate that multiple mechanisms drive the emergence of genes in grass- and pine-associated FFSC taxa examined and highlighted the diversity of molecular processes potentially underlying niche-specificity in these and other Fusarium species.

Adlay seed samples were collected from 3 adlay growing regions (Yeoncheon, Jeonnam and Eumseong regions) in Korea during 2012. Among all the samples collected, 400 seeds were tested for fungal occurrence by standard blotter and test tube agar methods and different taxonomic groups of fungal genera were detected. The most predominant fungal genera encountered were Fusarium, Phoma, Alternaria, Cladosporium, Curvularia, Cochliobolus and Leptosphaerulina. The occurrence of Fusarium species were 45.6% and based on the combined sequences of two protein coding genes, EF-1a, Beta-tubulin and phylogenetic analysis, 10 species were characterized as F. incarnatum (11.67%), F. kyushense (10.33%), F. fujikuroi (8.67%), F. concentricum (6.00%), F. asiaticum (5.67%), F. graminearum (1.67%), F. miscanthi (0.67%), F. polyphialidiom (0.33%), F. armeniacum (0.33%) and F. thapsinum (0.33%). The ability of these isolates to produce mycotoxins fumonisin (FUM) and zeralenone (ZEN) were tested by ELISA quantitative analysis method. The result revealed that fumonisin (FUM) was produced only by F. fujikuroi and zeralenone (ZEN) by F. asiaticum & F. graminearum. Mycotoxigenic species were then examined for their morphological characteristics to confirm their identity. Morphological observations of the species correlated well with their molecular identification and confirmed as F. asiaticum, F. fujikuroi and F. graminearum.

Fusarium head blight (FHB) is a wheat disease caused by fungi of the genus Fusarium. The aim of the study was to find relationships between the weather conditions in the experimental years and the locations and the amount of F. culmorum DNA and trichothecene genotypes, as well as the proportions between them. A three-year field experiment (2017, 2018 and 2019) was established at two locations (Poznań, Radzików). F. culmorum DNA was detected in all grain samples in an average amount of 20124 pg per 1 g of wheat DNA. The average amount of DNA from the 3ADON genotype was 4879 pg/μg and the amount of DNA from the NIV genotype was 3330 pg/μg. In the three experimental years, a large variability was observed in the coefficients of correlation between DNA concentrations and the FHB index, FDK, ergosterol, and the corresponding toxins. There were significant correlations between disease incidence, fungal biomass (quantified as the total amount of fungal DNA or DNA trichothecene genotypes) and toxins (DON, 3AcDON and NIV) concentrations. The 3ADON trichothecene genotype dominates over the NIV genotype (ratio 1.5); however, this varied greatly depending on environmental conditions.

In order to improve the yield of the therapeutic agent N-methylsansalvamide (SA), optimization of stationary liquid fermentation conditions was carried out on an endophytic strain, Fusarium sp. R1, at flask level. Using a One-Factor-At-a-Time approach, the fermentation conditions for SA production were determined as follows: fermentation time of 13 d, 24 oC, initial pH of 6.5, seed age of 24 h, inoculum size of 5.0% (v/v), loading volume of 50% (v/v), 20.0 g/L salinity, and sucrose, tryptone, and yeast extract were found to be the best sources of carbon and nitrogen. Using response surface methodology, the optimal medium compositions consist of 22.5 g/L sucrose, 16.5 g/L tryptone, and 0.024 g/L yeast extract. Verification tests suggested that the SA yield under these optimal conditions reached up to 536.77 ± 2.67 mg/L, which was increased by almost ten times the initial yield (54.05 ± 3.45 mg/L). The findings indicate that a high SA production yield can be achieved by static culture of strain R1 under the proper fermentation conditions using a low-cost medium. This study paves the way towards industrial-scale SA production by strain R1 for new drug development.

Fusarium species are agriculturally important fungi with a broad host range and can be found as endophytic, pathogenic, or opportunistic parasites in many crop plants. This study aimed to identify Fusarium species in bare-rooted, dormant plants in Turkish grapevine nurseries using molecular identification methods and assess their pathogenicity. Asymptomatic dormant plants were sampled from grapevine nurseries (43) in different regions of the country, and fungi were isolated from plant roots and internal basal tissues. The Fusarium isolates were identified by performing gene sequencing (TEF1-α, RPB2) and phylogenetic analyses. Pathogenicity tests were carried out by inoculating mycelial agar pieces of isolates onto the stem or conidial suspensions into the rhizosphere of vines (1103 Paulsen rootstock). Laboratory tests revealed that Fusarium species were highly prevalent in Turkish grapevine nurseries (41 out of 43). Gene sequencing and phylogenetic analyses unraveled that 12 Fusarium species (F. annulatum, F. brachygibbosum, F. clavum, F. curvatum, F. falciforme, F. fredkrugeri, F. glycines, F. nanum, F. nematophilum, F. nirenbergiae, F. solani, and Fusarium spp.) existed in the ready-to-sale plants. Some of these species (F. annulatum, F. curvatum and F. nirenbergiae) consistently caused wood necrosis of seedling stems, rotting of the basal zone and roots, and reduced root biomass. Although the other nine species also caused some root rot and root reduction, their virulence was not as severe as the pathogenic ones, and they were considered opportunistic parasites or endophytic species. This study is the first detailed investigation of Fusarium species in grapevine nurseries in Türkiye and emphasizes the need to consider pathogenic species in producing healthy grapevine seedlings.

Mycotoxin contamination of food and feed is a worldwide problem that needed to be addressed with highly efficient and biologically safe techniques. RNA interference (RNAi) is a natural mechanism playing an important role in different processes in eukaryotes, including regulation of gene expression, maintenance of genome stability, protection against viruses and others. Recently, RNAi-based techniques have been widely applied for the purposes of food safety and management of plant diseases, including those caused by mycotoxin-producing fungi. In this review, we summarize the current state-of-art of RNAi-based approaches for reducing aggressiveness of key toxigenic fungal pathogens and mycotoxin contamination of grain and its products. The ways of improving RNAi efficiency for plant protection and future perspectives of this technique, including progress in methods of double-stranded RNA production and its delivery to the target cells are also discussed.

Fish is a very healthy food that is a great source of proteins, vitamins, minerals, and essential fatty acids. For many people in developing nations, like Nigeria, smoked and dried fish serves as a significant source of protein and is an integral part of the traditional diet. However, smoked and dried fish are susceptible to fungal growth, especially when the storage conditions are not right, and it has been discovered that many products contain fungi that are easily visible on the surface. This study investigated the diversity and occurrence frequency of contaminated fungi species in dried fishes sold in local markets of Awka in Anambra state, Nigeria. Twenty-four fungal isolates with varying colony morphology were isolated from the various dried fish products. The contaminating species were further subjected to lactophenol cotton blue (LPCB) staining and Ten (10) Unique fungal morphology was detected. Carbohydrate utilization test was used to select unique isolates and further molecular identification showed successful PCR amplification of the isolates. The current study identified various fungal groups contaminating dried fish products sold in Awka Markets. The three dominant genera were the fungal genera' Aspergillus, Penicillium and Fusarium. These results suggest that most dried fish being sold in the Markets are contaminated with multiple fungi, posing a critical health risk to individuals. Thus, there is a need for attention to be paid to the packaging and storage of dried fish to lessen the likelihood of air exposure, lower water activity, and prevent microbiological growth on the dried fish.

Degnala is one of the main mycotoxicoses that affects buffaloes, and the fungus Fusarium spp. is the main causative agent. This disease is strongly related to the supply of rice straw to buffaloes and is considered endemic to regions where this crop is cultivated. Cases of the disease are concentrated in the winter, when conditions favour fungal growth in inadequately stored rice straw. Degnala is characterized by necrosis and gangrene of the extremities, including the tail, lower limbs, ears, tongue, muzzle and teats. The pelvic limbs are more affected than the thoracic limbs. A tortuous appearance of the tail is very common, and cracks or corneal loss of the hooves may occur, with exposure of the blades and even the bones. There is no diagnostic method for identifying the disease in animals other than the clinical and epidemiological criteria associated with fungal culture of rice straw samples. There is no treatment that neutralizes the effects of the toxins, and the treatment currently used is a palliative and supportive method consisting of pentasulfate solution, anti-Degnala liquid, 2% nitroglycerin ointment and broad-spectrum antibiotics for secondary infections. In addition, the management of drying straw and proper storage are essential for controlling this disease.

Fusarium solani KMZW-1, is recognized for its potential as a biocontrol agent against agricultural and forestry pests, particularly due to its compatibility with integrated pest management (IPM) strategies. This study aimed to investigate the complete genome of F. solani KMZW-1 and assess its pathogenicity against Bactrocera dorsalis. Whole genome sequencing revealed a genome size of 47,239,278 bp, comprising 27 contigs, with a GC content of 51.16% and fungus identified as F. solani KMZW-1. The genome completeness was assessed as 97.93% using BUSCO analysis, DFVF sequence identifier was Fusarium 0G092560.1, AntiSMASH analysis identified 29 gene clusters associated with secondary metabolite biosynthesis providing insights into the genetic basis of its pathogenic mechanisms and biocontrol potential. Comparative genomic analysis found 269 unique genes of F. solani KMZW-1 and the collinearity analysis exhibited a high degree of synteny with Fusarium solani-melongenae. The pathogenicity of F. solani KMZW-1 was assessed using concentrations ranging from 1×10^4 to 1×10^11 conidia/mL. Higher concentrations (1×10^10 to 1×10^11 conidia/mL), resulted in significantly increased cumulative mortality rates of B. dorsalis adults compared to the control group. Notably, the pathogenicity was higher in male adults than in females. Probit regression analysis yielded LogLC50 values as 5.662 for fe-male and 4.486 for male B. dorsalis adults. In summary, F. solani KMZW-1 exhibits strong insecti-cidal activity against B. dorsalis and shows potential as a biocontrol agent with IPM strategies. These findings provide robust genomic evidence supporting the use of F. solani KMZW-1 in managing against B. dorsalis populations.

Sunflower is one of the oil crops and its planting areas was listed behind Soybean, peanut and rapeseeds in China. It will encounter different kind of diseases during planting, thus limited the seeds yield. Fusarium species are the major threat to different crops nowadays, it is very important to identify the species caused diseases on different hosts, so that the potential control measures may be explored. In Inner mongolia, Sunflower Disk Rot (SDR) was found in different sunflower planting field, the average infection ratio around 8.50%, typically in some verieties, its infection ratio reach to 11.67%. The diseased receptacle were collected from different locations and Koch’s postulate was conducted. The results showed that Fusarium verticillioides was the main causal agent caused SDR. Fungicide toxicity tests shown that Fludioxonil and Flutolanil were screened out from six fungicides for controlling F. verticillioides, with EC50 values of 0.05 µg/mL (R = 0.9825) and 0.96 µg/mL (R = 0.9964) , respectively. This is the first report of SDR caused by F. verticillioides, and it will alert sunflower reseachers to include SDR in the disease list, so as to control sunflower diseases with integrated management strategy successfully.

A strategy to propose solutions to health-related problems recognizes that people, animals, and the environment are interconnected. Fusarium and Neocosmospora are an example of this interaction due to the capable of infecting plants, animals, and human. This review provides information on various aspects of these relations and proposes how to approach fusariosis with a One Health methodology. Here we give a framework to understand infection pathogenesis, through the epidemiological triad and explain how the broad utilization of fungicides in agriculture may play a role in the treatment of human fusariosis. We assess how plumbing systems and hospital environments might play a role as a reservoir for animal and human infections. We explain the role of antifungal resistance mechanism in both humans and agriculture. Our review emphasizes the importance of developing interdisciplinarity research studies where aquatic animals, plants, and human disease interactions can be explored through coordination and collaborative actions.

Fusarium stalk rot (FSR) caused by the Fusarium species complex is an economic threat to maize cultivation all over the world. We investigated the population structure and genetic diversity of Fusarium spp. obtained from five major maize-growing regions of India. The Tef-1α locus was used for phylogenetic analysis of geographically distinct isolates of Fusarium verticillioides, F. andiyazi, F. proliferatum, F. nygamai, and F. acutatum causing FSR. Geographical separation among four local populations contributed to 7.87% variance, whereas 92.12% of the variance was within popula-tions, suggesting a predominant influence of local adaptation or stochastic events like genetic drift within populations, with geographical separation playing a lesser but significant role in shaping genetic diversity. Genetic differentiation statistics between Fusarium species showed lower gene flow from haplotypes except in the population of F. acutatum and F. andiyazi having high GST values. In contrast, a high Kxy was reported, indicating elevated genetic differentiation between populations. The haplotype network analysis revealed the presence of eight distinct haplotypes which reflected pathogenic evolution and adaptive potential of Fusarium spp. The results offer a comprehensive framework for discussing the implications of genetic diversity in pathogen management and the evolutionary dynamics of the Fusarium spp. causing FSR of maize in the Indian subcontinent.

Growing acreage and changing consumer preferences cause increasing interest in the cereal products originating from organic farming. Lack of results of objective test, however, does not allow drawing conclusions about the effects of cultivation in the organic system and comparison to currently preferred conventional system. Field experiment was conducted in organic and conventional fields. Thirty modern cultivars of winter wheat were sown. They were characterized for disease infection including Fusarium head blight, seed sowing value, the amount of DNA of the six species of Fusarium fungi as well as concentration of ergosterol and trichothecenes in grain. The intensity Fusarium head blight was at a similar level in both systems. However, Fusarium colonization of kernels expressed as ergosterol level or DNA concentration was higher for the organic system. It did not reflect in an increased accumulation of trichothecenes in grain, which was similar in both systems, but sowing value of organically produced seeds was lower. Significant differences between analyzed cropping systems and experimental variants were found. The selection of the individual cultivars for organic growing in terms of resistance to diseases and contamination of grain with Fusarium toxins was possible. Effects of organic growing differ significantly from the conventional and grain obtained such way can be recommended to consumers. There are indications for use of particular cultivars bred for conventional agriculture in the case of organic farming, and the growing organic decreases plant stress resulting from intense fertilization and chemical plant protection.

Several studies explore in depth the biochemistry and genetics of the pigments present in Fusarium graminearum but there is a need to discuss about their relationship with the mold’s observable surface color pattern variation throughout its lifecycle. Furthermore, they require basic cataloguing and description of their major features known so far. Colors are a viable alternative to size measurement in growth studies. When grown on yeast extract agar (YEA) at 25 °C, F. graminearum initially exhibits a whitish mycelium, developing into a yellow-orange mold by the sixth day and then turning into wine-red. The colors are likely due to accumulation of the golden yellow polyketide aurofusarin and the red rubrofusarin, but the carotenoid neurosporaxanthin possibly play also a major role in the yellow or orange coloration. Torulene might contribute for red tones but it perhaps ends up being converted into neurosporaxanthin. Culmorin is also present but it does not contribute for the color, though it was initially isolated in pigment studies, and there is the 5-deoxybostrycoidin-based melanin, but it occurs mostly in the teleomorph’s perithecium. There is still a need to chemically quantify the pigments throughout the lifecycle, analyze their relationships and how much each impacts F. graminearum surface color.

Fusarium graminearum is a cereal pathogen responsible for economic losses worldwide every year. An understanding of its growth is key to control its infection, but current growth models are limited because their size-based approach provides little information about the mold's metabolism. Recently, a RGB (red, green and blue) imaging analysis demonstrated the predictability of F. graminearum color change as it grows in yeast extract agar (YEA). This study aimed to verify the same phenomenon in oats (a_w = 0.94, 0.97 and 0.99) and rice (a_w = 0.97, 0.98 and 0.99). Photos were taken using a professional camera and a smartphone (iPhone 6) after incubation and during the subsequent 16 days, and average RGB was quantified using ImageJ software. The photos showed very similar color variations, regardless of the type of grain or a_w. The mold first adopted a k-selection strategy by growing as a mycelium and then a r-selection strategy, increasing spore production. All RGB channels showed positive Pearson correlations between them (p < 0.001) and it was possible to design a model showing two lag phases, the first prior to a mycelial phase and the second prior to a sporular phase at the end of the experiment.

Dragon fruit (Hylocereus polyrhizus) constitutes an important economic industry in Guizhou Province, China; however, in recent years, stem rot in this region has become increasingly severe. Moreover, the pathogens responsible for stem rot in Guizhou and their sensitivity to fungicides remain elusive. Therefore, in this study, we aimed to determine the causative pathogens of stem rot in this region and analyze their sensitivity to fungicides. Twenty-four isolates were obtained from diseased tissues, from which H-4 and H-5 were selected and confirmed as pathogens. Based on the morphological characteristics of macroconidia, microconidia, and colony morphology, the polygenic phylogenetic tree constructed using internal transcribed spacer, elongation factor 1-alpha, and retinol-binding protein-2 gene fragments, along with carbon source metabolism using FF microplates, the two pathogens were identified as Fusarium oxysporum and F. concentricum respectively. In addition, the in vitro toxicity of eight fungicides against both pathogens were measured based on mycelium growth rate. The results showed that 75% trifloxystrobin·tebuconazole exhibited the strongest inhibitory effect on both isolates, with concentration for 50% of maximum effect values of 0.1262 µg/mL and 0.1385 µg/mL, respectively. This study identified two Fusarium spp. as the causative pathogens of stem rot in dragon fruit, with F. concentricum being reported for the first time, and demonstrated the best fungicide for them. These findings hold significant potential for guiding the effective treatment of stem rot in dragon fruit in Guizhou, China.

Fusarium species are one of the filamentous fungal taxa with the most pronounced impact on agri-cultural production and human health. The mycotoxins produced by pathogenic Fusarium not on-ly attack various plants including crops, causing various plant diseases that lead to reduced yields and even death, but also penetrate into the food chain of humans and animals to cause food poi-soning and consequent health hazards. Although sporadic studies have revealed some of the bio-synthetic pathways of Fusarium toxins, they are insufficient to satisfy the need for a comprehen-sive understanding of Fusarium toxin production. In this study, we focused on 35 serious patho-genic Fusarium species with available genomes and systematically analyzed the ubiquity of the dis-tribution of identified Fusarium and non-Fusarium derived fungal toxin biosynthesis gene clusters (BGCs) in these species through the mining of core genes and comparative analysis of correspond-ing BGCs. Additionally, novel sesterterpene synthases and PKS_NRPS clusters were discovered and analysed. This work is the first to systematically analyze the distribution of related mycotoxin biosynthesis in pathogenic Fusarium species. These findings enhance the knowledge of mycotoxin production and provide a theoretical grounding for the prevention of fungal toxin production us-ing biotechnological approaches.

Size-based fungal growth studies have limitations. For example, the growth in size stops in closed systems once it reaches the borders and poorly describes the metabolic status, especially in the stationary phase. This might lead mycotoxin studies to unrealistic results. Color change could be a viable alternative as pigments are results of the mold’s metabolic activity. This study aimed to verify the possibility of using gray values and the RGB system to analyze the growth of Fusarium graminearum. It consisted color and area measurement using the ImageJ software for specimens grown in yeast extract agar (YEA). The results suggest the usability of color and gray values as reliable tools to analyze the growth of F. graminearum.

Dalbergia sissoo (shisham), an important timber yielding multipurpose tree species of the Indian subcontinent, has been afflicted with large scale mortality due to wilt in natural forests and plantations, causing huge economic losses. Fusarium solani f. sp. dalbergiae (Fsd) has been identified as one of the causal organisms for wilt disease in D. sissoo. Present study comprises in vitro screening of ten selected genotypes of D. sissoo against two strains of Fsd in a dual culture set up under axenic condition. Callus and plantlets of ten genotypes of host plant were multiplied in vitro and were inoculated with conidial suspension of two strains of Fsd at three concentrations; 1× 10¹, 1× 10³, and 1× 10⁵ conidia/ml. Gnotobiotic evaluation of dual culture set up shows variations among D. sissoo genotypes in their response towards in vitro Fsd infection; and two genotypes (14 and 66) exhibited resistance against the pathogen strains. Callus of genotypes 14 and 66 significantly restricted the fungal mycelium growth whereas callus of remaining eight genotypes were completely infested by Fsd mycelium within 9 days. Similarly, plantlets of genotype 14 and 66, had lesser disease severity and remained green, and had fewer necrotic lesions in the roots whereas plantlets of remaining eight genotypes died within 15 days.

Deoxynivalenol (DON) is a well-known mycotoxin, responsible for outbreaks of gastrointestinal disorders in Japan. Fusarium graminearum, a parasite of cereal crops, produces this toxin and this is one of the reasons why it is important to understand its metabolism. It is possible to predict the mold’s color change and the quantity of DON synthesized throughout its lifecycle. Furthermore, a_w has been found to affect the amount of DON. This study aimed to analyze the potential of F. graminearum surface color as a predictor of DON concentration at a_w = 0.94, 0.97 and 0.99. Thus, 36 specimens were incubated at 25 °C, 12 at each a_w. After 4, 8, 12 and 16 days, 3 specimens from each a_w were collected for color analysis and DON quantification. For color analysis, photos were taken and red, green and blue (RGB) channels were measured on ImageJ software. DON was quantified through liquid chromatography (HPLC). Color changes were only observed at a_w = 0.99 because at lower a_w the molds presented high growth of white mycelium. Yet, DON increased in all cases. It was only possible to relate the colors with DON concentration at a_w = 0.99, where they presented inverse proportionality.

Fusarium graminearum is a dangerous pathogen of the cereals producing mycotoxins (trichothecene and zearalenone) harmful for human and animal health. There were evaluated sixteen winter wheat varieties for their response in conditions of natural infection with F. graminearum in the epidemic year 2019, being well known that accumulation of mycotoxins (DON, ZON and T-2) is induced by different biotic and abiotic factors. Field plot was organized in Latin rectangle randomized with three replicates. For all evaluated wheat varieties were collected field data (incidence, severity and infection degree of the fungus F. graminearum) and laboratory data (mycotoxins concentration in grains) that have been processed using the software JASP (Version 0.14) for descriptive statistics, and exploratory factor analysis (EFA). Microsoft Excel 2019 was used to calculate Pearson’s correlation coefficients. The results showed negative corelation between plants’ density and F. graminearum attack frequency. Positive correlations were found between DON and T-2 and between DON and fungus attack intensity. This work highlights that during a F. graminearum epidemic year some of the most influential factors in the contamination with harmful mycotoxins (DON, ZON and T-2) are: plants density, frequency of the attack on ear, diseased ears and attack intensity on ears.

Fusarium head blight (FHB) can cause contamination of cereal grain with mycotoxins. Winter wheat is also infected with FHB. It is more resistant than durum wheat to head infection and less than other small grain cereals. The aim of this study was to identify winter wheat lines that combine low head infection and kernel damage with low toxin contamination. Resistance to FHB of 27 winter wheat lines and cultivars was evaluated over a three-year experiment established in two locations. At the anthesis stage, heads were inoculated with Fusarium culmorum isolates. The FHB index was scored and the percentage of Fusarium-damaged kernels (FDKs) assessed. The grain was analysed for type B trichothecenes (deoxynivalenol and derivatives, nivalenol) and zearalenone content. The average FHB index was 12.9%. The proportion of FDK was 6.9% (weight) and 8.5% (number). An average content of deoxynivalenol amounted to 3.543 mg/kg and nivalenol to 2.115 mg/kg. In total, it was 5.804 m/kg of type B trichothecenes. The zearalenone content in the grain was 0.214 mg/kg. Relationships between FHB index, FDK and mycotoxin contents were highly significant for wheat lines; however, they were stronger for FDK versus mycotoxins. Breeding lines combing all types of FHB resistance were found, and five of them had resistance similar to that of wheat lines with the Fhb1 gene.

Oats are a nutrient rich cereal used for animal feed and growing in human consumption. This cereal can be affected by Fusarium spp., causing the disease Fusarium Head Blight. This disease is caused mainly by species within the Fusarium graminearum species complex, and are also responsible for producing mycotoxins that are harmful to humans and animals. This study aimed to investigate fungal diversity in Brazilian oat samples, focusing on the Fusarium sambucinum species complex and the presence of type B trichothecenes (deoxynivalenol and its derivatives, and nivalenol) from two different regions; Paraná (PR) and Rio Grande do Sul (RS). The isolated fungi from oat grains were identified as species from the genera: Fusarium, Phoma and Alternaria. The majority of Fusarium isolates belonged to the Fusarium sambucinum species complex; identified as F. graminearum s.s., F. meridionale and F. poae. In the RS region, F. poae was the most frequent fungus, while FGSC was the most frequent in the PR region. The majority of F. graminearum s.s. isolates were of the 15-ADON genotype, while some 3-ADON genotypes were identified; however, F. meridionale and F. poae were all of the NIV genotype. Mycotoxin analysis revealed that 92% and 100% of the samples from PR and RS were contaminated with type B trichothecenes, respectively. The oats from PR were predominantly contaminated with DON, whereas NIV was predominant in oats from RS. Analysis showed that 24% of the samples were contaminated with DON at levels higher than Brazilian regulations. Co-contamination of DON, its derivatives and NIV was observed in 84% and 57.7% of the samples from PR and RS, respectively. The results provide new information on Fusarium contamination in Brazilian oats, highlighting the importance for further studies on mycotoxins.

Deoxynivalenol (DON) is a mycotoxin produced by Fusarium spp. that causes Fusarium head blight (FHB) disease in cereal crops. Ingestion of food contaminated with DON poses serious human health complications. However, the DON cytotoxicity has been mostly deduced from animal studies. In this study, we used the nematode Caenorhabditis elegans (C. elegans) as a tractable animal model to dissect the toxic effect of DON. Our results indicate that DON reduces the fecundity and lifespan of C. elegans. The real-time RT-PCR analysis showed that DON upregulates innate immunity-related genes including C17H12.8 and K08D8.5 encoding PMK-1 (mitogen activated protein kinase-1)-regulated immune effectors, and F35E12.5 encoding a CUB-like domain-containing protein. Furthermore, our RNAseq data demonstrate that out of ~ 17,000 C. elegans genes, 313 are upregulated and 166 were downregulated by DON treatment. Among the DON-upregulated genes, several are ugt genes encoding UDP-glucuronosyl transferase (UGTs) which are known to be involved in chemical detoxification. The three upregulated genes, F52F10.4 (oac-32), C10H11.6 (ugt-26) and C10H11.4 (ugt-28) encoding the O-acyltransferase homolog, UGT26 and UGT 28, respectively, are shown to contribute to DON tolerance by RNAi bacterial feeding experiment. The results of this study provide insights to the targets of DON cytotoxicity and potential mitigation measures.

Fungal keratitis (FK) is an invasive infection of the cornea primarily associated with Aspergillus and Fusarium species. FK is treated empirically with a limited selection of topical antifungals with varying levels of success. Though clinical infections are typically characterized by a dense network of mature mycelium, traditional models used to test antifungal susceptibility of FK isolates exclusively evaluate susceptibility in fungal cultures derived from asexual spores known as conidia. The purpose of this study was to characterize differences in fungal response when topical antifungal treatment is initiated at progressive phases of fungal development. We compared efficacy of voriconazole and luliconazole against in vitro cultures of A. flavus and F. keratoplasticum at 0, 24, and 48 h of fungal development. A porcine cadaver corneal model was used to compare antifungal efficacy of voriconazole and luliconazole in ex vivo tissue cultures of A. flavus and F. keratoplasticum at 0, 24, and 48 h of fungal development. Our results demonstrate phase-dependent susceptibility of both A. flavus and F. keratoplasticum to both azoles in vitro as well as ex vivo. We conclude that traditional antifungal susceptibility testing with conidial suspensions does not correlate with fungal susceptibility in cultures of a more advanced developmental phase. A revised method of antifungal susceptibility testing that evaluates hyphal susceptibility may better predict fungal response in the clinical setting where treatment is often delayed until days after the initial insult.

The soil-borne pathogens, particularly Fusarium oxysporum f. sp. niveum (FON) and southern root-knot nematode (RKN, Meloidogyne incognita) are the major threat to watermelon production in the south-eastern United States. The role of soil micronutrients on induced resistance (IR) to plant diseases is well-documented in soil-based mediums. However, soil-based mediums limit the determination of the role(s) of individual micronutrients in IR. In this manuscript, we utilized hydroponics to assess the effect of controlled application of micronutrient, including iron (Fe), manganese (Mn), and zinc (Zn) on the expression of some IR genes (PR1, PR5, and NPR1 from salicylic acid (SA) pathway, and VSP, PDF, and LOX genes from jasmonic acid (JA) pathway) in watermelon seedlings upon inoculation with either FON or RKN or both. Plants were treated with higher (3X) or lower (0.5X) concentrations of micronutrients in Steiner solution (X= standard dose of micronutrient) for 7 days in a hydroponics system under greenhouse conditions. A subset of micronutrient-treated plants was inoculated (on the 8th day of micronutrient application) with FON and RKN (single and mixed). The expression of the IR genes in treated and control samples were evaluated using qRT-PCR. Although, significant phenotypic differences were not observed with respect to the severity of wilt symptoms or RKN galling with any of the micro-nutrient treatments within the 30 day-experimental-period, differences in the induction of IR genes were observed. However, the level of gene expression varied with sampling period, type and concentration of micro-nutrients ap-plied, and pathogen-inoculation. In the absence of pathogens, no significant changes were observed in the expression level of IR genes on 7th day of micronutrient treatment. However, pathogen inoculation affected the expression levels of the IR genes at 3-day post-inoculation. In FON inoculated plants, PDF was upregulated in high Fe treatment, whereas in RKN inoculated plants, low Mn treatment resulted in up-regulation of VSP. In the case of mixed inoculation with FON and RKN, the plants with low Zn treatment resulted in the upregulation of PR1. These observations suggest that the type and concentration of micronutrients in watermelon may potentially induce systemic resistance against FON and RKN through SA and JA pathways.

Zucchini (Cucurbita pepo L.) is an important crash crop for producing seeds in Inner Mongolia, China. In August 2021, the harvest season of seed-producing zucchini, fruit rot symptom (cv. Jindi No.1) was found in the Wuyuan region. The incidence of infected fruits of zucchini ranged from 10% to 30% and symptom consisted in 4 to 5 cm diameter lesions where the fruit was in contact with the soil. Ten isolations from six symptomatic fruits were preliminary classified as Fusarium species based on its morphological characters in potato dextrose agar. Phylogenetic tree was constructed by concatenating TEF-1α and RPB2 gene and exhibited the isolates clustered together with the Fusarium solani f. sp. cucurbitae sequences. Pathogenicity tests revealed that two isolates caused over 3 cm diameter lesions on the fruits while negative-controls remained asymptomatic. The colonies reisolated from the leasions were morphologically identical to the original isolates, thus fulfilling Koch’s postulates. Results demonstrated that F. solani f. sp. cucurbitae was the causal agent of fruit rot observed in the field. To our knowledge, this is the first report of fruit rot of seed-producing zucchini caused by F. solani f. sp. cucurbitae in Inner Mongolia, China.

Fusarium oxysporum f. sp. lycopersici (Fol) causes vascular wilt disease in tomato. Upon colonization of the host, Fol secretes many small effector proteins into the xylem sap to facilitate infection. Besides known SIX (Secreted In Xylem) proteins, the identity of additional effectors that contribute to Fol pathogenicity remains largely unexplored. We have performed a deep RNA-sequencing analysis of Fol race 2-infected tomato, used the sequence data to annotate a published genome assembly generated via PacBio SMRT sequencing of the Fol race 2 reference strain Fol4287, and analysed the resulting transcriptome to identify Fol effector candidates among the newly annotated genes. We examined the Fol-infection expression profiles of all 13 SIX genes present in Fol race 2 and identified 27 new candidate effector genes that were likewise significantly upregulated upon Fol infection. Using Agrobacterium-mediated transformation, we tested the ability of 22 of the new candidate effector genes to suppress or induce cell death in leaves of Nicotiana benthamiana. One effector candidate designated Fol-EC19, encoding a secreted guanyl-specific ribonuclease, was found to trigger cell death and two effector candidates designated Fol-EC14 and Fol-EC20, encoding a glucanase and a secreted trypsin, respectively, were identified that can suppress Bax-mediated cell death. Remarkably, Fol-EC14 and Fol-EC20 were also found to suppress I-2/Avr2- and I/Avr1-mediated cell death. Using the yeast secretion-trap screening system, we showed that these three biologically-active effector candidates each contain a functional signal peptide for protein secretion. Our findings provide a basis for further understanding the virulence functions of Fol effectors.

The effects of soluble silicon fertilization on monocots and dicots have been widely studied. However, little is known regarding its effects on protecting epiphytes against insect and fungal pests. The efficacy of silicon fertilizer to reduce damage by thrip pest complexes, namely: Thrips palmi Karny, Frankliniella occidentalis Pergande, Chaetanaphothrips orchidii Moulton, and Chaetanaphothrips signipennis Bagnall (Thysanoptera: Thripidae), and fungal pathogens: Botrytis cinerea Persoon (Helotiales: Sclerotiniaceae) and Fusarium spp. Link (Hypocreales: Nectriaceae) was examined during a nine-month greenhouse trial in Hawaii. The trial assessed yield, quality, and pest damage across three common varieties of dendrobiums. All replicates received additional soluble silicon fertilizer applications alternating weekly between soil drench and foliar (50 mg Si/plant) applications. Yield, quality, and spray length, pest damage, plant vigor, SPAD, and leaf temperature were measured. Data were analyzed using a generalized linear model (glm) with repeated measures followed by post-hoc pair-wise comparisons in R, version 4.3.1. Treatment effects were significant at P

The FTF (Fusarium Transcription Factor) gene family is composed of two members (FTF1 and FTF2) with high sequence homology. Both genes encode Zn(II)2Cys6 binuclear zinc finger transcription factors involved in the modulation of virulence in the F. oxysporum species complex (FOSC). While FTF1 is a multicopy gene exclusive of highly virulent strains of FOSC and is located in the accessory genome, FTF2 is a single copy gene, located in the core genome, and well conserved in all filamentous ascomycete fungi, except yeast. The involvement of FTF1 in the colonization of the vascular system and regulation of the expression of SIX effectors has been stablished. To address the role of FTF2 we generated and characterized null FTF2 mutants in a F. oxysporum f. sp. phaseoli weakly virulent strain and analyzed them together with the equivalent mutants formerly obtained in a highly virulent strain. The results obtained highlight a role for FTF2 as a negative regulator of the production of macroconidia and demonstrate that it is required for full virulence and positive regulation of SIX effectors. In addition, gene expression analyses provide compelling evidence that FTF2 is involved in the regulation of hydrophobins likely required for plant colonization.

Fusarium head blight (FHB) disease is extensively distributed worldwide. This disease damages grain quality and reduces yield. The detection of this disease in a high throughput way is crucial to planters and breeders. Our study focused on developing a method for processing wheat color images and accurately detecting disease areas using deep learning and image processing techniques. The color images of wheat at the milky stage were collected and processed to construct datasets, which were used to retrain a deep convolutional neural network model using transfer learning. Testing results showed that the model can detect spikes, and the coefficient of determination of the number of spikes between the manual count and the detection was 0.80. The model was assessed, and the mean average precision for the testing dataset was 0.9201. On the basis of the results of spike detection, a new color feature was applied to obtain the gray image of each spike. Then, a modified region growing algorithm was implemented to segment and detect the diseased areas of each spike. Results show that the region growing algorithm performs better than K-means and Otsu’s method in segmenting the FHB disease. Overall, this study demonstrates that deep learning techniques enable the accurate detection of FHB in wheat using color images, and the proposed method can effectively detect spikes and diseased areas, thereby improving the efficiency of FHB detection.

The aim of the study was to determine the presence Fusarium species and mycotoxins in winter wheat grain in Poland. Grain samples from different locations in Poland in 2009 and 2010 were analysed for the content of biomass of Fusarium species and mycotoxins. In 2009 biomass of F. graminearum and F. poae was present in all samples, F. culmorum in 82% of samples, F. avenaceum in 55% of samples. F. sporotrichioides, F. tricinctum and F. equiseti were found only in individual samples. F. langsethiae was not detected. In 2010, five Fusarium species were detected with the exception of F. sporotrichioides. The highest content of biomass was found for F. graminearum followed by F. avenaceum, F. poae and F. langsethiae. The amount of F. culmorum biomass was very low. The most frequently occurring species was F. poae and F. graminearum. In 2009, deoxynivalenol was detected in all samples. In 2010, the average content of deoxynivalenol was lower than in 2009. Nivalenol was detected at very low concentration in both years. Significant correlations between content of F. graminearum biomass and deoxynivalenol concentration in grain and between content of F. poae biomass and nivalenol concentration in grain in 2009 were found. The most important finding of this study was that main Fusarium species infecting wheat kernels in Poland in both years was F. graminearum. The amount of biomass of F. graminearum was the highest in both years. It was present in the most samples. The other frequently detected species was F. poae, which in 2010 appeared in more samples than F. graminearum. However, the amount of F. poae biomass was lower. F. culmorum, species that was previously dominating as wheat pathogen in Poland, was found less frequently than F. graminearum. The amount of biomass of this species was the lowest in 2010.

Fusarium graminearum causes head blight in wheat and corn, and produces chemicals harmful for humans and other animals. It is important to understand how it grows in order to prevent outbreaks. There are 3 well-known growth models for microorganisms and they seem applicable to molds: linear, Gompertz and Baranyi. This study aimed to see which could better represent F. graminearum growth. Three replicates were grown in yeast extract agar (YEA) for 20 days. The Feret’s radius was measured in ImageJ software, and then related to the models. Linear model was the most closely correlated to the actual growth. Thus, considering that it was the most representative of the reality and it is easier to use, it seems to be the best logical choice for F. graminearum growth studies.

Seven agronomic factors (crop season, farming system, harvest date, moisture, county, oat variety, and previous crop) were recorded for 202 oat crops grown across Ireland, and samples were analysed by LC-MS/MS for four major Fusarium mycotoxins: deoxynivalenol (DON), zearalenone (ZEN), T-2 toxin and HT-2 toxin. Type A trichothecenes were present in 62% of crops, with 7.4% exceeding European regulatory limits. DON (6.4%) and ZEN (9.9%) occurrences were rela-tively infrequent, though one and three samples were measured over their set limits respectively. Overall, the type of farming system and the previous crop were the main factors identified to significantly influence mycotoxin prevalence or concentration. Particularly, adherence to an organic farming system and growing oats after a previous crop of grass were found to decrease contamination by type A trichothecenes. These are important findings and may provide valuable insights for many other types of cereals crops as Europe moves towards a much greater organic based food system.

Chalcones are a class of flavonoids possessing a variety of biological activities, including antimicrobial and antioxidant effects. Therefore, chalcones and their derivatives have potential use in the control of postharvest diseases. In this work, the effects of 2ʹ,4ʹ-dichloro-chalcone on the in vitro growth and in vivo pathogenicity of F. tricinctum and T. roseum were investigated. The results showed that 100 µM of 2ʹ,4ʹ-dichloro-chalcone strongly inhibited mycelial growth and conidial production of F. tricinctum (32.3%) and T. roseum (65.2%) in vitro. This treatment also significantly inhibited the expansion of potato dry rot from F. tricinctum (48.6%) and apple rot spot from T. roseum (36.2%). The incubation of F. tricinctum and T. roseum conidia with this agent for 2 h increased their cell membrane permeability by 25% and 22.5%, respectively and conidial membrane permeability by 41.7 and 65.4%, respectively. This treatment also significantly inhibited the total respiration rate and activated the cyanide-resistant respiratory pathway in both pathogens. Protein immunoblotting showed that the treatment increased alternative oxidase (AOX) levels after 4 h in F. tricinctum and T. roseum by 52.76% and 39.13%, respectively. Conclusion: 100 µM 2ʹ,4ʹ-dichloro-chalcone significantly inhibited mycelial growth and spore production in F. tricinctum and T. roseum in vitro and reduced their pathogenicity in postharvest potato and apple crops. Further analysis indicated that 2ʹ,4ʹ-dichloro-chalcone disrupted the integrity of cell membranes and the mitochondrial respiratory electron transport chain which leaded to ROS burst and oxidative stress, AOXs were induced directly or indirectly, and the cyanide resistant respiratory pathway was activated. The in vitro growth and pathogenicity of F. tricinctum and T. roseum were inhibited.

Cotton (Gossypium spp.) is the most important fibre crop worldwide. Black root rot and Fusarium wilt are two major diseases of cotton caused by soil-borne Berkeleyomyces rouxiae and Fusarium oxysporum f. sp. vasinfectum (Fov), respectively. Phenotyping plant symptoms caused by soil-borne pathogens has always been a challenge. To increase the uniformity of infection, we adapted a seedling screening method that directly uses liquid cultures to inoculate the plant roots and the soil. Four isolates each of B. rouxiae and Fov were collected from cotton fields in Australia and were characterised for virulence on cotton under controlled plant growth conditions. While the identity of all four B. rouxiae isolates were confirmed by multi-loci sequencing, only two of them were found to be pathogenic on cotton, suggesting variability in the ability of isolates of this species to cause disease. The four Fov isolates were phylogenetically clustered together with the other Australian Fov isolates and displayed both external and internal symptoms characteristic of Fusarium wilt on cotton plants. Furthermore, the isolates appeared to induce varied levels of plant disease severity indicating differences in their virulence on cotton. To contrast the virulence of the Fov isolates, four putatively non-pathogenic Fusarium oxysporum (Fo) isolates collected from cotton seedlings exhibiting atypical wilt symptoms were assessed for their ability to colonise cotton host. Despite the absence of Secreted In Xylem genes (SIX6, SIX11, SIX13 and SIX14) characteristic of Fov, all four Fo isolates retained the ability to colonise cotton and induce wilt symptoms. This suggests that slightly virulent strains of Fo may contribute to the overall occurrence of Fusarium wilt in cotton fields. Findings from this study will allow better distinction to be made between plant pathogens and endophytes and allow fungal effectors underpinning pathogenicity to be explored.

Water-miscible metalworking fluids (MWFs) are utilized in a variety of metal removal and forming operations. For end-use, formulation concentrates are diluted in water, creating conditions conducive to microbial growth and metabolism, possibly compromising the fluid’s integrity and mechanically obstructing filters or piping systems. Metalworking machines offer additional habitats on surfaces that are in permanent or temporary contact with MWFs. For that reason, biocides have been incorporated into concentrates for years, but legal constraints will restrain their use in the future. While bacterial contamination of MWFs is well documented, fungal contamination is often overseen and infrequently reported in the literature. In this study, we report fungal prevalence in in-use MWFs sampled worldwide over 10 years and we are convinced that presence of fungi is the norm rather than the exception. In addition, we evaluated the inhibitory effect of fungicides on fungal growth, sporulation and spore viability using traditional culture-dependent methods and flow cytometry. In essence, we show that the effectiveness of these fungicides is limited and dependent on the chemical construction of the fluid. We think that the ecology created by water-diluted MWFs is of higher importance than the anti-fungal activity of single components.

Fusarium Head Blight (FHB) is a destructive disease affecting the grain yield and quality of wheat, barley, rye and triticale. Developing varieties with genetic resistance is integral to successfully managing FHB. However, significant knowledge gap exists in the genetic diversity present in triticale for FHB resistance. This information is critical for breeding new varieties of triticale as its production continues to increase. In the present study, a set of 298 winter triticale accessions from a worldwide collection were screened for their type-2 FHB resistance in an artificially inoculated misted nursery with high levels of inoculum density. Most of the triticale accessions were susceptible to FHB, and only 8% of accessions showed resistance in the field nursery screening. The resistant accessions identified in the nursery screening were selected and further screened for three years in greenhouse conditions. Seven accessions were found to show robust FHB resistance over the three years of greenhouse testing. Thirteen accessions showed significantly lower levels of Deoxynivalenol accumulation when compared to the susceptible triticale control. The accessions identified in the study will be useful in triticale and wheat breeding programs for enhancing FHB resistance and reducing DON accumulation.

Plant pathogenic fungi are the most important cause of losses in agriculture. In the face of the overuse of synthetic fungicides, natural products are an encouraging alternative to control these plant pathogenic fungi. In this study, we tested the antifungal capacity of metabolites produced by strains of the genus Streptomyces against the plant pathogenic fungi Alternaria alternata, Botrytis cinerea and Fusarium oxysporum. In addition, we investigated the feasibility of applying these metabolites in a postharvest setting for the control of B. cinerea on cherry tomato fruits. The results revealed the ability of the strains, especially Streptomyces netropsis, to inhibit the growth of plant pathogenic fungi and, in some cases, the sporulation of these fungi. In addition, we observed an interesting phenomenon in which Streptomyces strains physically encapsulated the mycelium of the fungi, which contributed to their inhibition. Finally, the protective activity of S. netropsis extract on Cherry tomato fruits infected with B. cinerea was demonstrated. In conclusion, this research opens the door to studies on the understanding of physical microbial interactions in natural environments and holds promise in agriculture, especially in the protection of agricultural products during the post-harvest stage as a safer and more sustainable alternative.

Members of the Fusarium oxysporum species complex (FOSC) has the capacity to specialize into host-specific pathogens known as formae speciales through horizontal gene transfer between pathogenic and endophytic individuals. To this day, the origin of these formae speciales and the genetic determinants dictating the switch from endophytic to pathogenic Fusarium oxysporum (Fox) are still unknown. F. oxysporum f. sp. vanillae (Fov), member of FOSC, is the causal agent of root and stem rot disease, representing the main phytosanitary problem in vanilla plantations worldwide. Here we analyzed the RNA-seq libraries resulting from the interaction vanilla-Fov at early and late stages of the infection, and what we initially identified as control in a previous study, detecting the presence of Fox endophytes. We identified virulence, hypervirulence, sporulation, conidiation, necrosis, and production of fusaric acid as key processes taking place during Fov-vanilla interaction. Through comparison with endophytic Fox, we found that Fov can infect vanilla thanks to the presence of pathogenicity islands and genomic regions associated with supernumerary chromosomes. These play a central role as carriers of genes involved with pathogenic activity and could have being obtained by Fov through horizontal gene transfer. We also found that, unlike other pathogenic members of FOSC, Fov do not use Secreted in Xylem proteins (SIX) to infect vanilla.

Molecular identification via Internal Transcribed Spacer region (nrDNA-ITS) sequencing of Fusarium spp. isolates from wheat originated from Stara Zagora region, were performed for the first time in Bulgaria. А total of 60 wheat samples (Triticum aestivum) were morphologically identified at the genus level as Fusarium spp. in advance. The rDNA-ITS region of all isolates was successfully amplified and the PCR products obtained were directly sequenced. After a comparison of detected sequences with NCBI database, members of three different fungal genera (Fusarium, Chaetomium, and Alternaria) were identified. Among Fusarium isolates, the F. tricinctum was prevailing, followed by F. poae. A total of three isolate F. proliferatum, F. graminearum and F. equiseti were presented with a single probe. The lowest genetic distance (0.004) was detected between F. tricinctum isolates. On the base of genetic distances, fungal isolates were grouped in two main clusters – one comprising F. tricinctum isolates and F. proliferatum, and second including F. equiseti, F. graminearum and F. poae. It could be concluded that the rDNA-ITS genome region of the genus Fusarium may be used as a suitable marker of early detection, accurate and reliable identification of Fusarium spp. contamination of wheat. The timely and accurate information would assist in the selection of appropriate approaches for control of fusarium infections and possible mycotoxins contamination of agricultural production.

Abstract: In Australia adoption of reduced tillage and stubble retention cropping systems to conserve soil moisture by growers has seen an increase in the prevalence of the disease Fusarium crown rot (FCR) caused by the stubble-borne fungal pathogen Fusarium pseudograminearum. Durum wheat is particularly susceptible to FCR expressing significant yield and quality losses in the presence of infection. Increasing rates of nitrogen (N) application at sowing exacerbates FCR. However, to achieve the desired grain protein and quality suited to pasta manufacture, N application is necessary and this creates a dilemma for growers. The purpose of this study was to investigate the effects of FCR infection in the presence of different N fertiliser application rates in durum wheat varieties on yield and technological quality. Two durum varieties were evaluated at the same location over two seasons (2020 and 2021). These seasons were characterised by being wetter than normal and showed different responses to FCR and N application. In general, the 2021 season showed better responses to applied N with no impact of FCR. The FCR inoculation while it resulted in significant infection, it had no impacts on yield or grain quality in 2021 while in 2020 yield was reduced and only gluten index was reduced. Jandaroi was found to hold its kernel vitreosity at all protein levels obtaining the premium grade while DBA Lillaroi did not and would suffer downgrade if N application was insufficient. However, higher N application rates needed to achieve 12% + protein saw a reduction in dough strength with Jandaroi holding its dough strength much better than DBA Lillaroi. This was related to the lower glutenin/gliadin (Gli/Glu) ratio in response to applied N at sowing in Jandaroi which helped retain kernels with high vitreousness. This suggests genetics plays an important role in a genotype response to N fertilisation and should be considered when selecting a variety where higher premium grades are desirable.

Decoctions, macerations and fumigations of the stem bark and wood of Terminalia brownii Fresen. are used in traditional medicine for fungal infections and as pesticides on field crops and in traditional granaries in Sudan. In addition, T. brownii is commonly used for protecting wooden houses and furniture. Therefore, using agar disc diffusion and macrodilution methods, eight extracts of various polarities from the stem wood and bark were screened for their growth inhibitory effects against filamentous fungi commonly causing fruit, vegetable and grain decay, as well as infections in the immunocompromised host. Ethyl acetate extracts of the stem wood and bark gave the best antifungal activities, with MIC values of 250 µg/ml against Nattrassia mangiferae and Fusarium verticillioides, and 500 µg/ml against Aspergillus niger and Aspergillus flavus. Aqueous extracts gave almost as potent effects as the ethyl acetate extracts against the Aspergillus and Fusarium strains, and were slightly more active than the ethyl acetate extracts against Nattrassia mangiferae. Thin layer chromatography, RP-HPLC-DAD and tandem mass spectrometry (LC-MS/MS), were employed to identify the chemical constituents in the ethyl acetate fractions of the stem bark and wood. The stem bark and wood were found to have a similar qualitative composition of polyphenols and triterpenoids, but differed quantitatively from each other. The stilbene derivatives, cis- (3) and trans- (4) resveratrol-3-O-β-galloylglucoside, were identified for the first time in T. brownii. Moreover, methyl-(S)-flavogallonate (5), quercetin-7-β-O-di-glucoside (8), quercetin-7-O-galloyl-glucoside (10), naringenin-4`-methoxy-7-pyranoside (7), 5,6-dihydroxy-3`,4`,7-tri-methoxy flavone (12), gallagic acid dilactone (terminalin) (6), a corilagin derivative (9) and two oleanane type triterpenoids (1) and (2) were characterized. Our results justify the traditional uses of macerations and decoctions of T. brownii stem wood and bark for crop and wood protection and demonstrate that standardized extracts could have uses for the eco-friendly control of plant pathogenic fungi in African agroforestry systems. Likewise, our results justify the traditional uses of these preparations for the treatment of skin infections caused by filamentous fungi.

Watermelon fusarium wilt caused by Fusarium oxysporum f.sp. niveum (FON) is a destructive soil-borne disease throughout the world leading to serious economic losses and limit watermelon production. Cuminic acid, extracted from the seed of Cuminum cyminum L., belongs to benzoic acid analogues. In this study, the median effective concentration (EC₅₀) values for cuminic acid in inhibiting mycelial growth of FON was 22.53μg/mL. After treatment with cuminic acid, mycelial morphology was seriously influenced; cell membrane permeability and glycerol content were increased markedly, but pigment and mycotoxin (mainly fusaric acid) were significantly decreased. Synthesis genes of bikaverin and fusaric acid both were down regulated compared with the control confirmed by quantitative RT-PCR. In greenhouse experiments, cuminic acid at all concentrations displayed significant bioactivities against FON. Importantly, significant enhancement of activities of SOD, POD, CAT and decrease of MDA content after cuminic acid treatment in watermelon leaves were observed in vivo. These indicated that cuminic acid not only showed high antifungal activity, but also could enhance the self-defense system of the host plant. Above all, cuminic acid showed the potential as a biofungicide to control FON.

A plant’s early response to pathogen stress is a vital indicator of its disease resistance. In order to study the response mechanism of resistant and susceptible flax cultivars to Fusarium oxysporum f. sp. lini (Foln), we applied RNA-sequencing to analyze transcriptomes of flax with Foln 0.5, 2 and 8 hours post inoculation (hpi). We found a significant difference in the number of differential expression genes (DEGs) between resistant and susceptible flax clutivars. The number of DEGs in the Fusarium-resistant cultivar increased dramatically at 2 hpi, and a large number of DEGs participated in the Fusarium-susceptible cultivar response to Foln infection 0.5 hpi. GO enrichment analysis determined that the up-regulated DEGs of both flax cultivars were enriched such as oxidoreductase activity and oxidation-reduction process. At the same time, the genes involved in diterpenoid synthesis were up-regulated in resistant cultivar, while those involved in extracellular region, cell wall and organophosphate ester transport were down-regulated in susceptible cultivar. KEGG enrichment analysis showed the genes encoded WRKY 22 and WRKY33 which involved in MAPK signaling pathway were up-regulated expressed in S-29 and down-regulated expressed in R-7, negatively regulated the disease resistance of flax; The genes encoded Hsp 90 family which in involved in plant pathogen interaction pathway were up-regulated in R-7 and down-regulated in S-29, which positively regulated the disease resistance of flax; The genes encoded MYC2 transcription factor and TIFY proteins which involved in plant hormone signaling pathway were up-regulated in R-7, and regulated the jasmonic acid metabolism of flax and the signal transduction of plant hormones. Meanwhile seven regulatory genes with the most correlation were screened out, Among Lus10025000.g and Lus10026447.g regulated other genes expressed both in plant hormone signal transduction pathway and MAPK signal pathway. In conclusion, these findings will facilitate further studies on the function of these candidate genes in flax of response to Fusarium stress, and the breeding of disease-resistant flax cultivar.

RNA-viruses of the genera Ambivirus, Mitovirus, Sclerotimonavirus, and Partitivirus were found in a single isolate of Fusarium graminearum. The genomes of the mitovirus, sclerotimonavirus, and partitivirus were assigned to previously described viruses, whereas the ambivirus genome putatively represents a new species, named Fusarium graminearum ambivirus 1 (FgAV1). To investigate the effect of mycoviruses on the fungal phenotype, spontaneous loss of mycoviruses during meiosis and transmission of mycoviruses into a new strain by anastomosis were used to obtain isogenic F. graminearum strains with and without mycoviruses. Notable effects observed in mycovirus-harboring strains were (i) suppression of the synthesis of trichothecene mycotoxins and their precursor trichodiene, (ii) suppression of the synthesis of the defense compound aurofusarin, (iii) stimulation of the emission of 2-methyl-1-butanol and 3-methyl-1-butanol, and (iv) increased attractiveness of fungal mycelia for fungivorous collembolans. Increased attractiveness of mycovirus-infected filamentous fungi to animal predators opens new perspectives on the ecological implications of infection of fungi with viruses.

Gibberella ear rot (GER) caused by Fusarium graminearum (teleomorph Gibberella zeae) is one of the most destructive diseases in maize that severely reduces grain yield and contaminates several potential mycotoxins. However, few efforts had been devoted to dissect the genetic basis of maize GER resistance. In the present study, a genome-wide association study (GWAS) was conducted in a maize association panel consisting of 303 diverse inbred lines. The phenotypes of GER severity were evaluated using kernel bioassay across multiple time points in the laboratory. Then, three models including fixed and random model circulating probability unification model (FarmCPU), general linear model (GLM) and mixed linear model (MLM), were conducted simultaneously in GWAS to identify single-nucleotide polymorphisms (SNPs) significantly associated with GER resistance. A total of four individual significant association SNPs with the phenotypic variation explained (PVE) ranging from 3.51 to 6.42% were obtained. Interestingly, the peak SNP (PUT-163a-71443302-3341) with the greatest PVE value, was co-localized in all models. Subsequently, 12 putative genes were captured from the peak SNP that several of these genes were directly or indirectly involved in disease resistance. Overall, these findings contribute to understand the complex plant-pathogen interactions in maize GER resistance. The regions and genes identified herein provide a list of candidate targets for further investigation, in addition to the kernel bioassay that can be used for evaluating and selecting elite germplasm resources with GER resistance in maize.

Cereal crops are affected by one of the most devastating diseases worldwide, known as Fusarium head blight (FHB), with Fusarium graminearum being the most isolated causal pathogen. Another species associated to this disease is Fusarium poae. This species has been considered a relatively weak pathogen compared to F. graminearum, but its importance has increased for its occurrence in cereal grains worldwide. Considering the advantages of using B. distachyon as a plant model and the importance of F. poae in crops, our study aimed to evaluate the potential use of Brachypodium as a plant model to evaluate the compatible interaction with F. poae. For this, 12 Brachypodium spp. accessions from different countries were inoculated with a selected F. poae set of isolates. Disease severity, conidial quantification, fungal DNA biomass, and nivalenol quantification were assessed. The results showed a compatible interaction between Brachypodium accessions and Fusarium poae, which allowed the use of the model plant for future plant-pathogen interaction studies.

Fusarium spp. are commonly associated with the root rot complex of soybean (Glycine max). Previous surveys identified six common Fusarium species from Manitoba, including F. oxysporum, F. redolens, F. graminearum, F. solani, F. avenaceum and F. acuminatum. This study aimed to determine their pathogenicity, assess host resistance and evaluate the genetic diversity of Fusarium spp. isolated from Canada. Pathogenicity was tested on two cultivars, ‘Akras’ (moderate resistance) and ‘B150Y1’ (susceptible), under greenhouse conditions. The virulence of the fungal isolates varied, with root rot severities ranging from 1.5 to 3.3 on a 0-4 scale. The six species were used to screen a panel of 20 Canadian soybean cultivars for resistance in a greenhouse. Cluster and principal component analyses were conducted based on the same traits as for the pathogenicity study. Two of the cultivars, ‘P15T46R2’ and ‘B150Y1’, were consistently found to be tolerant to F. oxysporum, F. redolens, F. graminearum and F. solani. To investigate the incidence and prevalence of Fusarium spp. in Canada, fungi were isolated from 106 soybean fields surveyed across Manitoba, Saskatchewan, Ontario and Quebec. Two-hundred twenty-one Fusarium isolates were identified, with phylogenetic analyses indicating diversity of the isolates in the major soybean production regions of Canada.

Fusarium circinatum is an important global pathogen of pine trees. Genome plasticity has been observed in different isolates of the fungus, but no genome comparisons are available. To address this gap, we sequenced and assembled to chromosome level five isolates of F. circinatum. These genomes were analysed together with previously published genomes of F. circinatum isolates FSP34 and KS17. Multi-sample variant calling identified a total of 461683 micro variants (SNPs and small indels) and a total of 1828 macro structural variants of which 1717 were copy number variants and 111 were inversions. Variant density was higher on sub-telomeric regions of chromosomes. Variant annotation revealed that genes involved in transcription, transport, metabolism and transmembrane proteins were overrepresented in gene sets affected by high impact variants. A core genome representing genomic elements conserved in all the isolates and a non-redundant pangenome representing all genomic elements is presented. Whole genome alignments showed that an average of 93% of the genomic elements are present in all isolates. The results of this study reveal that some genomic elements are not conserved within the isolates and some variants are high impact. The described genome-scale variations will help inform novel disease management strategies against the pathogen.

Fungal keratitis (FK) is a serious ocular infection that often poses significant diagnostic and therapeutic dilemmas. This study aimed to examine the causes, clinical characteristics, outcomes, and prognostic factors of FK in the UK. All culture-positive and culture-negative presumed FK (with complete data) that presented to Queen’s Medical Centre, Nottingham, and Queen Victoria Hospital, East Grinstead, between 2011 and 2020 were included. A total of 117 patients (n=117 eyes) with FK were included in this study. The mean age was 59.0±19.6 years (range, 4-92 years) and 51.3% patients were female. Fifty-three fungal isolates were identified from 52 (44.4%) culture-positive cases, with Candida spp. (33, 62.3%), Fusarium spp. (9, 17.0%), and Aspergillus spp. (5, 9.4%) being the most common organisms. Ocular surface disease (60, 51.3%), prior corneal surgery (44, 37.6%), and systemic immunosuppression (42, 35.9%) were the three most common risk factors. Hospitalisation for intensive treatment was required for 95 (81.2%) patients, with a duration of 18.9±16.3 days. Sixty-six (56.4%) patients required additional surgical interventions for eradicating the infection. Emergency therapeutic/tectonic keratoplasty was performed in 29 (24.8%) cases, though 13 (44.8%) of them failed at final follow-up. The final corrected-distance-visual-acuity (CDVA) was 1.67±1.08 logMAR. Multivariable logistic regression analyses demonstrated increased age, large infiltrate size (>3mm), and poor presenting CDVA (<1.0 logMAR) as significant negative predictive factors for poor visual outcome (CDVA of <1.0 logMAR) and poor corneal healing (>60 days of healing time or occurrence of corneal perforation requiring emergency keratoplasty; all p<0.05). In conclusion, FK represents a difficult-to-treat ocular infection that often results in poor visual outcome, with a high need for surgical interventions. Innovative treatment strategies are urgently required to tackle this unmet need.

Fusarium culmorum is a major pathogen of grain crops. Infected plants accumulate deoxynivalenol (DON), 3-acetyl-deoxynivalenol (3-ADON), or nivalenol (NIV), which are mycotoxins of the trichothecene B group. These toxins are also produced by F. graminearum species complex. New trichothecenes structurally similar to trichothecenes B but lacking the carbonyl group on C-8, designated NX toxins, were recently discovered in atypical isolates of F. graminearum from North America. Only these isolates and a few strains of a yet to be characterized Fusarium species from South Africa are known to produce NX-2 and other NX toxins. Here we report that among 20 F. culmorum strains isolated from maize, wheat, and oat in Europe and Asia over a period of 70 years, 18 strains produced NX-2 simultaneously with 3-ADON and DON or NIV. Rice cultures of strains producing 3-ADON accumulated NX-2 in amounts corresponding to 2-8% of 3-ADON (1.2 - 36 mg/kg). A strain producing NIV accumulated NX-2 and NIV at comparable amounts (13.6 and 10.3 mg/kg, respectively). In F. graminearum, producers of NX-2 possess a special variant of cytochrome P450 monooxygenase encoded by TRI1 that is unable to oxidize C-8. In F. culmorum, producers and nonproducers of NX-2 possess identical TRI1; the reason for the production of NX-2 is unknown. Our results indicate that production of NX-2 simultaneously with trichothecenes B is a common feature of F. culmorum.

2,5-Di(hydroxymethyl)furan (DHMF) is a high-value chemical block than can be synthesized from 5-hydroxymethylfurfural (HMF), a platform chemical that results from the dehydration of biomass-derived carbohydrates. In this work, the HMF biotransformation capability of different Fusarium species was evaluated and F. striatum was selected to produce DHMF. The effects of the inoculum size, glucose concentration and pH of the media over DHMF production were evalu-ated by a 23 factorial design. A substrate feeding approach was found suitable to overcome the toxicity effect of HMF towards the cells when added at high concentrations (>75 mM). The pro-cess was successfully scaled-up at bioreactor scale (1.3 L) with excellent DHMF production yields (95%) and selectivities (98%). DHMF was purified from the reaction media with high recovery and purity by organic solvent extraction with ethyl acetate.

Fusarium wilt of banana is a devastating disease that has decimated banana production worldwide. Host resistance to Fusarium oxysporum f. sp. cubense (Foc), the causal agent of this disease, is genetically dissected in this study using two Musa acuminata ssp. malaccensis segregating populations segregating for Foc Tropical (TR4) and Subtropical (STR4) race 4 resistance. Marker loci and trait association using 11 SNP-based PCR markers allowed the candidate region to be delimited to a 12.9 cM genetic interval corresponding to a 959 kb region on Chromosome 3 of ‘DH-Pahang’ reference assembly v4. Within this region, there is a cluster of pattern recognition receptors, namely leucine rich repeat ectodomain containing receptor-like protein kinases, cysteine-rich cell wall associated protein kinases, and leaf rust 10 disease-resistance locus receptor-like proteins positioned in an interspersed arrangement. Their transcript levels were rapidly upregulated in the resistant but not in susceptible F2 progenies at the onset of infection. This suggests that one or several of these genes may control resistance at this locus. To confirm the segregation of single-gene resistance, we generated an inter-cross between the resistant parent ‘Ma850’ and a susceptible line ‘Ma848’, to show that the STR4 resistance co-segregated with marker ‘28820’ at this locus. Finally, an informative SNP marker 29730 allowed the locus specific resistance to be assessed in a collection of diploid and polyploid banana plants. Out of the 60 lines screened, 22 lines were predicted to carry resistance at this locus, including lines known to be TR4 resistant, such as ‘Pahang’, ‘SH-3362’, ‘SH-3217’, ‘Ma-ITC0250’, and ‘DH-Pahang/CIRAD 930’. Additional screening in the International Institute for Tropical Agriculture’s collection suggests that the dominant allele is common among in the elite ‘Matooke’ NARITA hybrids, as well as in other triploid or tetraploid hybrids derived from East African highland bananas. Fine-mapping and candidate gene identification will allow characterization of molecular mechanisms underlying TR4 resistance. The markers developed in this study can now aid the marker-assisted selection of TR4 resistance in breeding programs around the world.

Sarocladium and Fusarium species are commonly identified as causal agents of rice sheath rot disease worldwide. However, limited knowledge exists about their genetic, pathogenic, and toxigenic diversity in sub-Saharan African (SSA) countries, where an increasing incidence of this disease has been observed. This study aimed at identifying, characterizing, as well as assessing the genetic, pathogenic, and toxigenic diversity of the pathogens associated with rice sheath rot disease in Mali, Nigeria, and Rwanda. In this study seventy fungal isolates were obtained from rice plants displaying disease symptoms on research and farmer’s fields in Mali, Nigeria, and Rwanda. Thus, an extensive comparative analysis was conducted to assess their genetic, pathogenic, and toxigenic diversity. The Fusarium spp. were characterized using the translation elongation factor (EF-1α) region, while a concatenation of Internal Transcribed Spacer (ITS) and Actin-encoding regions were used to resolve Sarocladium species. Phylogenetic analysis revealed four Fusarium species complexes. The dominant complex in Nigeria was the Fusarium incarnatum-equiseti species complex (FIESC), comprising F. hainanense, F. sulawesiense, F. pernambucatum, and F. tanahbumbuense, while F. incarnatum was found in Rwanda. The Fusarium fujikuroi species complex (FFSC) was predominant in Rwanda and Mali, with species such as F. andiyazi, F. madaense, and F. casha in Rwanda, and F. annulatum and F. nygamai in Mali. F. marum was found in Nigeria. Furthermore, Fusarium oxysporum species complex (FOSC) members, F. callistephi and F. triseptatum, were found in Rwanda and Mali, respectively. Two isolates of F. acasiae-mearnsii, belonging to the Fusarium sambucinum species complex (FSAMSC) were obtained in Rwanda. Isolates of Sarocladium which were previously classified in three phylogenetic groups were resolved into three species which are: attenuatum, oryzae and sparsum. S. attenuatum was dominant in Rwanda, while S. oryzae and S. sparsum were found in Nigeria. Also, the susceptibility of FARO44, a rice cultivar released by Africa Rice Centre (AfricaRice) was tested against isolates from the four Fusarium species complexes and the three Sarocladium species. All isolates evaluated could induce typical sheath rot symptoms albeit with varying disease development levels. In addition, liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used to determine variation in the in vitro mycotoxins of the Fusarium species. Regional differences were observed in the in vitro mycotoxins profiling. Out of the forty-six isolates tested, nineteen were able to produce one to four mycotoxins. Notably, very high zearalenone (ZEN) production was specific to the two F. hainanense isolates from Ibadan-Nigeria, while Fusarium nygamai isolates from Mali produced high amounts of fumonisins. To the best of our knowledge, it seems this study is the first to elucidate genetic, pathogenic, and toxigenic diversity of Fusarium species associated with the rice sheath rot disease complex in selected countries in SSA.

Main pathogens affecting carob (Ceratonia siliqua) tree in the Mediterranean basin are described in this overview. The most widespread diseases periodically occurring on carob orchards are powdery mildew (Pseudoidium ceratoniae) and cercospora leaf spot (Pseudocercospora ceratoniae). The causal agents of so-called “black leaf spots” (e.g. Pestaliotiopsis, Phyllosticta and Septoria spp.) are responsible for similar symptoms to above mentioned foliar diseases but are reported on carob orchards with a negligible frequency. Likewise, canker and branch diebacks caused by fungal species belonging to Botryosphaeriaceae are almost never recorded. Among wood diseases that may compromise old or previously damaged carob specimens, “brown cubical heart rot” caused by Laetiporus sulphureus is the most widespread and recurrent issue; this pathogen is also well-known for edible fruit bodies that are also appreciated for pharmaceutical and industrial purposes. On the other hand, “white decays” caused by Fomes and Ganoderma species are less common and reported for the first time in this review. Like-gall protuberances on twigs at uncertain aetiology or tumours on branches associated to Rhyzobium radiobacter are described although these symptoms are seldom detected as it happens also for necrotic leaf spots caused by Pseudomonas syringae pv. ciccaronei. A list of worldwide pathogens not yet recorded but at high-risk potential of introduction in Italian carob producing area is also provided. Finally, some concerns are addressed to new phytopathogenic fungi vectored by invasive Xylosandrus compactus ambrosia beetle. Comprehensively, all described pathogens could potentially in near future become limiting factors for carob production, because they could be favoured by high-density orchards, increasing global network of trade exchanges and high frequency through which extreme events of climate changes globally occur. Thus, symptoms and signs, causal agents, epidemiology and whenever applicable recommendations for disease prevention and management are provided in this review.

Repeat-Induced Point mutations (RIP) serves as a genome defence mechanism that impedes the deleterious consequences of repeated motifs such as transposable elements in fungi. Genomic regions with RIP are biased for adenosine and thymine transitions and the cumulative influence of RIP is thought to have a considerable impact on genome composition. We investigated the impact of RIP on localized genomic regions and whole-genome sequences for representatives of the pine pathogen, Fusarium circinatum. We set out to determine the intraspecific variation in acquired RIP and the role of RIP in the development of diverse F. circinatum sub-genomic compartments. The results of the study show that the AT-enriched sub-genomic compartment accounts for ca. 97% of the calculated RIP and was further prominent in both core and accessory genomic regions. However, more extensive RIP was observed in the accessory sub-compartment and more variable regions of the genome. Regions with RIP indicated increased intrinsic curvature of the DNA which may influence DNA-protein interactions and may promote constitutive heterochromatin formation. The results show that RIP is an important source of functional novelty and genome variation. RIP contributes to the evolution of the genetic landscape and differentiation of diverse sub-genomic compartments of this important fungal pathogen.

Trichoderma atroviride is a strong necrotrophic mycoparasite antagonizing and feeding on a broad range of fungal phytopathogens. It further beneficially acts on plants by enhancing growth in root and shoot and inducing systemic resistance. Volatile organic compounds (VOCs) are playing a major role in all those processes. To date, T. atroviride IMI 206040 and T. atroviride P1 are among the most frequently studied T. atroviride strains and hence are used as model organisms to study mycoparasitism and photoconidiation. However, there are no studies available, which systematically and comparatively analyzed putative differences between these strains regarding their light-dependent behavior. We therefore explored the influence of light on conidiation and the mycoparasitic interaction as well as the light-dependent production of VOCs in both strains. Our data show that in contrast to T. atroviride IMI 206040 conidiation in strain P1 is independent of light. Furthermore, significant strain-and light-dependent differences in the production of several VOCs between the two strains became evident, indicating that T. atroviride P1 could be a better candidate for plant protection than IMI 206040.

The increase in arrivals of new forest pests highlights the need for effective phytosanitary legislation and measures. This paper introduces legislation targeted at prevention and management of potential introductions of forest pests and pathogens. A summary is given on plant health regulations on a global and regional level with detailed information on the situation in the European Union (EU). The current and new European legislation is discussed and a particular focus is given on eradication and contingency plans for Fusarium circinatum.