Detection of 2-ethyl-1-hexanol and its modulating effect in biofilm of Fusarium oxysporum.

In immunocompetent individuals, Fusarium spp. stands out as the causative agent of onychomycosis, among the non-dermatophyte molds. Despite evidence indicating that Fusarium oxysporum organizes itself in the form of a biofilm causing onychomycosis, there is little literature on the etiopathogenesis of the biofilm on the nail, specifically the signaling molecules present, known as quorum sensing (QS). Thus, this study detected the presence of a molecule related to QS from the ex vivo biofilm of F. oxysporum on human nail and investigated its effect on preformed biofilm in vitro. The detection and physicochemical characterization of a QS molecule, from the extracellular matrix (ECM), was carried out by Fourier transform infrared (FTIR) spectroscopy with an attenuated total reflectance (ATR) accessory and by headspace gas chromatography coupled to mass spectrometry (GC-MS) analyses. Determination of viable cells, cell activity, total biomass, ECM components and scanning electron microscopy (SEM) were performed to evaluate the influence of the QS molecule on the in vitro biofilm of F. oxysporum. The beginning, in the ex vivo biofilm of F. oxysporum on human nails, the volatile organic compound 2-ethyl-1-hexanol (2EH) was detected as a component of QS. Thereafter in vitro analyses, synthetic 2EH was able to modulate the biofilm by stimulating its filament, increasing total biomass and ECM production in terms of total carbohydrates, but with a reduction in total proteins and nucleic acids. We thus evidence, for the first time, the presence of 2EH in the biofilm of F. oxysporum, developed on the human nail, and the in vitro action of this compound as a QS molecule.

Shift in the rhizosphere soil fungal community associated with root rot infection of Plukenetia volubilis Linneo caused by Fusarium and Rhizopus species.

BACKGROUND: Plukenetia volubilis Linneo is an oleaginous plant belonging to the family Euphorbiaceae. Due to its seeds containing a high content of edible oil and rich in vitamins, P. volubilis is cultivated as an economical plant worldwide. However, the cultivation and growth of P. volubilis is challenged by phytopathogen invasion leading to production loss. METHODS: In the current study, we tested the pathogenicity of fungal pathogens isolated from root rot infected P. volubilis plant tissues by inoculating them into healthy P. volubilis seedlings. Metagenomic sequencing was used to assess the shift in the fungal community of P. volubilis rhizosphere soil after root rot infection. RESULTS: Four Fusarium isolates and two Rhizopus isolates were found to be root rot causative agents of P. volubilis as they induced typical root rot symptoms in healthy seedlings. The metagenomic sequencing data showed that root rot infection altered the rhizosphere fungal community. In root rot infected soil, the richness and diversity indices increased or decreased depending on pathogens. The four most abundant phyla across all samples were Ascomycota, Glomeromycota, Basidiomycota, and Mortierellomycota. In infected soil, the relative abundance of each phylum increased or decreased depending on the pathogen and functional taxonomic classification. CONCLUSIONS: Based on our results, we concluded that Fusarium and Rhizopus species cause root rot infection of P. volubilis. In root rot infected P. volubilis, the shift in the rhizosphere fungal community was pathogen-dependent. These findings may serve as a key point for a future study on the biocontrol of root rot of P. volubilis.

Resistance risk, resistance mechanism and the effect on DON production of a new SDHI fungicide cyclobutrifluram in Fusarium graminearum.

Fusarium head blight in wheat is caused by Fusarium graminearum, resulting in significant yield losses and grain contamination with deoxynivalenol (DON), which poses a potential threat to animal health. Cyclobutrifluram, a newly developed succinate dehydrogenase inhibitor, has shown excellent inhibition of Fusarium spp. However, the resistance risk of F. graminearum to cyclobutrifluram and the molecular mechanism of resistance have not been determined. In this study, we established the average EC50 of a range of F. graminearum isolates to cyclobutrifluram to be 0.0110 µg/mL. Six cyclobutrifluram-resistant mutants were obtained using fungicide adaptation. All mutants exhibited impaired fitness relative to their parental isolates. This was evident from measurements of mycelial growth, conidiation, conidial germination, virulence, and DON production. Interestingly, cyclobutrifluram did not seem to affect the DON production of either the sensitive isolates or the resistant mutants. Furthermore, a positive cross-resistance was observed between cyclobutrifluram and pydiflumetofen. These findings suggest that F. graminearum carries a moderate to high risk of developing resistance to cyclobutrifluram. Additionally, point mutations H248Y in FgSdhB and A73V in FgSdhC1 of F. graminearum were observed in the cyclobutrifluram-resistant mutants. Finally, an overexpression transformation assay and molecular docking indicated that FgSdhBH248Y or FgSdhC1A73V could confer resistance of F. graminearum to cyclobutrifluram.

Metconazole inhibits fungal growth and toxin production in major Fusarium species that cause rice panicle blight.

Rice panicle blight (RPB) caused by various Fusarium spp. is an emerging disease in the major rice-growing regions of China. Epidemics of this disease cause significant yield loss and reduce grain quality by contaminating panicles with different Fusarium toxins. However, there is currently no registered fungicide for the control of RPB in China. The 14α-demethylation inhibitor (DMI) fungicide metconazole has been shown to be effective against several Fusarium spp. that cause wheat head blight, wheat crown rot and maize ear rot. In this study, we investigated the specific activity of metconazole against six Fusarium spp. that cause RPB. Metconazole significantly inhibited mycelial growth, conidium formation, germination, germ tube elongation and major toxin production in Fusarium strains collected from major rice-growing regions in China, as well as disrupting cell membrane function by inhibiting ergosterol biosynthesis. Greenhouse experiments indicated a significant reduction in blight occurrence and toxin accumulation in rice panicles treated with metconazole. Overall, our study demonstrated the potential of metconazole for managing RPB and toxin contamination, as well as providing insight into its bioactivities and modes of action of metconazole against distinct Fusarium spp.

Unravelling Species Diversity and Pathogenicity of Fusarium spp. Associated with Soybean Leaf and Root in Heilongjiang Province, China.

Soybean (Glycine max L.) holds significant global importance and is extensively cultivated in Heilongjiang Province, China. Soybean can be infected by Fusarium species, causing root rot, seed decay, stem rot, and leaf blight. In 2021 to 2022, a field survey of soybean diseases was carried out in 11 regions of Heilongjiang Province, and 186 soybean leaves with leaf blight symptoms and 123 soybean roots with root rot symptoms were collected. Unexpectedly, a considerable number of Fusarium isolates were obtained not only from root samples but also from leaf samples. A total of 584 Fusarium isolates (416 from leaves and 168 from roots) were obtained and identified as 18 Fusarium species based on morphological features and multilocus phylogenetic analyses with tef1 and rpb2 sequences. Fusarium graminearum and Fusarium sp. 1 in FOSC were the dominant species within soybean leaf and root samples, respectively. Pathogenicity tests were conducted for all Fusarium isolates on both soybean leaves and roots. Results showed that F. graminearum, F. ipomoeae, F. citri, F. compactum, F. flagelliforme, F. acuminatum, and F. sporotrichioides were pathogenic to both soybean leaves and roots. F. solani, F. avenaceum, F. pentaseptatum, F. serpentinum, F. annulatum, and Fusarium sp. 1 in FOSC were pathogenic to soybean roots, not to leaves. To our knowledge, this is the first study to thoroughly investigate soybean-associated Fusarium populations in leaves and roots in Heilongjiang Province.

Characterizing the Pathogenicity and Mycotoxin Production Capacity of Fusarium spp. Causing Root Rot of Angelica sinensis in China.

Root rot is a very destructive soil-borne disease, which severely affects the quality and yield of Angelica sinensis in major planting areas of Gansu Province, China. Twelve Fusarium strains were identified from root rot tissue and infected soil in the field by comparing each isolate strain internal transcribed spacer, translation elongation factor 1-α sequence and RNA polymerase second largest subunit gene with the sequences of known fungal species in the NCBI database. Of these isolates, four were F. acuminatum, followed by three F. solani, two F. oxysporum, and one each of F. equiseti, F. redolens, and F. avenaceum. Under greenhouse conditions, pathogenicity testing experiment was carried out using five strains: two F. acuminatum, one F. solani, one F. oxysporum, and one F. equiseti. Among them, the incidence of F. acuminatum-induced root rot on A. sinensis was 100%; hence, it was the most aggressive. Liquid chromatography was used to show that F. acuminatum could produce neosolaniol (NEO), deoxynivalenol, and T-2 toxins. Of these, the level of NEO produced by F. acuminatum was high compared with the other two toxins. By isolating Fusarium spp. and characterizing their toxin-producing capacity, this work provides new information for effectively preventing and controlling A. sinensis root rot in the field as well as improving the quality of its medicinal materials.

Diversity and Pathogenicity of Fusarium Root Rot Fungi from Canola (Brassica napus) in Alberta, Canada.

Root rot disease poses a significant threat to canola (Brassica napus), underscoring the need for a comprehensive understanding of its causal agents for more effective disease mitigation. The composition and diversity of fungal pathogens associated with root rot of canola in Alberta, Canada, were evaluated from plant tissue samples collected in 2021 and 2022. The study revealed Fusarium spp. as the predominant pathogens found in almost all surveyed fields. Fusarium avenaceum, F. redolens, and F. solani were among the most frequently recovered species. Greenhouse trials confirmed their pathogenicity, with F. avenaceum and F. sporotrichioides found to be particularly aggressive. Additionally, F. sporotrichioides and F. commune were identified for the first time as canola root rot pathogens. Inoculation with isolates of most species resulted in significant reductions in seedling emergence, plant height, and shoot and root dry weights. Analysis of translation elongation factor 1-α (TEF-1α) and internal transcribed spacer (ITS) sequences confirmed the identity of the Fusarium spp., while concatenating the ITS and TEF-1α sequences enabled improved species differentiation. Geographic and year effects did not influence fungal diversity or aggressiveness, as determined by principal component analysis. This study emphasized the high diversity and impact of Fusarium spp. in causing canola root rot.

Inhibitory Effect of Sorbus aucuparia Extracts on the Fusarium proliferatum and F. culmorum Growth and Mycotoxin Biosynthesis.

Fungal infections are among the most common diseases of crop plants. Various species of the Fusarium spp. are naturally prevalent and globally cause the qualitative and quantitative losses of farming commodities, mainly cereals, fruits, and vegetables. In addition, Fusarium spp. can synthesize toxic secondary metabolites-mycotoxins under high temperature and humidity conditions. Among the strategies against Fusarium spp. incidence and mycotoxins biosynthesis, the application of biological control, specifically natural plant extracts, has proved to be one of the solutions as an alternative to chemical treatments. Notably, rowanberries taken from Sorbus aucuparia are a rich source of phytochemicals, such as vitamins, carotenoids, flavonoids, and phenolic acids, as well as minerals, including iron, potassium, and magnesium, making them promising candidates for biological control strategies. The study aimed to investigate the effect of rowanberry extracts obtained by supercritical fluid extraction (SFE) under different conditions on the growth of Fusarium (F. culmorum and F. proliferatum) and mycotoxin biosynthesis. The results showed that various extracts had different effects on Fusarium growth as well as ergosterol content and mycotoxin biosynthesis. These findings suggest that rowanberry extracts obtained by the SFE method could be a natural alternative to synthetic fungicides for eradicating Fusarium pathogens in crops, particularly cereal grains. However, more research is necessary to evaluate their efficacy against other Fusarium species and in vivo applications.

Essential oils of thyme (Thymus vulgaris) and oregano (Origanum vulgare) as an alternative for the control of pesticide-resistant Fusarium spp. in quinoa seeds.

BACKGROUND: The phytopathogenic genus Fusarium can cause damage such as root and stem rot in economically important crops, with significant implications. To seek a sustainable method for controlling this phytopathogen in seeds, the antifungal activity of essential oils (EOs) from thyme (Thymus vulgaris) and oregano (Origanum vulgare) was evaluated against isolates of F. graminearum, F. equiseti, F. culmorum and F. oxysporum originating from quinoa (Chenopodium quinoa) crops in the Boyacá (Colombia). RESULTS: Initially, the effectiveness of commercial fungicides against the mentioned phytopathogenic fungi was evaluated. Upon verifying that these isolates exhibited high resistance to these compounds, the EOs were assessed as a potential control alternative. A disk diffusion assay demonstrated complete in vitro inhibition of the growth of the evaluated phytopathogens when undiluted EOs were used. Subsequently, the minimum inhibitory concentration (MIC) of these oils was determined using the agar well diffusion technique, revealing a MIC of 10 and 1 µL mL-1 for thyme and oregano oil, respectively. Following this, the antifungal activity of the EOs applied to quinoa seeds was evaluated, and germination indices were measured as an indirect indicator of their toxicity. CONCLUSION: Despite both EOs successfully inhibiting microbial growth in the seeds, it was also found that thyme EO at 100 µL mL-1 and oregano EO at 10 µL mL-1 inhibited seed emergence and germination. However, lower concentrations exhibited a reduction in fungal population without affecting these germination indices. Therefore, it is suggested that the use of these compounds has potential in the treatment and disinfection of quinoa seeds. © 2024 Society of Chemical Industry.

Changing climate, shifting mycotoxins: A comprehensive review of climate change impact on mycotoxin contamination.

Climate change (CC) is a complex phenomenon that has the potential to significantly alter marine, terrestrial, and freshwater ecosystems worldwide. Global warming of 2°C is expected to be exceeded during the 21st century, and the frequency of extreme weather events, including floods, storms, droughts, extreme temperatures, and wildfires, has intensified globally over recent decades, differently affecting areas of the world. How CC may impact multiple food safety hazards is increasingly evident, with mycotoxin contamination in particular gaining in prominence. Research focusing on CC effects on mycotoxin contamination in edible crops has developed considerably throughout the years. Therefore, we conducted a comprehensive literature search to collect available studies in the scientific literature published between 2000 and 2023. The selected papers highlighted how warmer temperatures are enabling the migration, introduction, and mounting abundance of thermophilic and thermotolerant fungal species, including those producing mycotoxins. Certain mycotoxigenic fungal species, such as Aspergillus flavus and Fusarium graminearum, are expected to readily acclimatize to new conditions and could become more aggressive pathogens. Furthermore, abiotic stress factors resulting from CC are expected to weaken the resistance of host crops, rendering them more vulnerable to fungal disease outbreaks. Changed interactions of mycotoxigenic fungi are likewise expected, with the effect of influencing the prevalence and co-occurrence of mycotoxins in the future. Looking ahead, future research should focus on improving predictive modeling, expanding research into different pathosystems, and facilitating the application of effective strategies to mitigate the impact of CC.

Stalk rot species diversity and molecular phylogeny associated with diseased maize in India.

Stalk rot disease is a major constraint in maize production and till date reported to be caused by two to three species of phytopathogenic fungi but, in our present study, we disclose the first report of stalk rot is caused by complex species of phytopathogens, which belongs to five different genera. Therefore, to substantiate these findings, a total of 105 diseased samples of maize were collected from 21 different locations in six different geographical locations of India from which 48 isolates were used for the research study. Morphological features such as pigmentation, colony color, type of mycelium and pattern of mycelium was examined using macro and microscopic methods. A total of 11 different spp. of pathogens belonging to the five different genera: Fusarium verticillioides (56.25%), F. equiseti (14.5%), F. andiyazi (6.25%), F. solani (2.08%), F. proliferatum (2.08%), F. incarnatum (2.08%), Lasidioplodia theobrame (6.25%), Exserohilum rostrtum (4.16%), Nigrospora spp. (4.16%). and Schizophyllum commune (2.08%) were identified by different housekeeping genes (ITS, TEF-1α, RPB2 and Actin). Fusarium verticillioides, F. equiseti and F. andiyazi were major pathogens involved in stalk rot. This is the first report on F. proliferatum, F. solani, F. incarnatum, Lasidioplodia theobrame, Exserohilum rostrtum, Nigrospora spp. and Schizophyllum commune causing stalk rot of maize and their distribution in the different states of India. Studies on population dynamics of PFSR will enhance the understanding of pathogen behavior, virulence, or its association with different pathogens across India, which will facilitate the development of resistant maize genotypes against the PFSR.

Identification and pathogenicity analysis of Fusarium spp. on peach in China.

BACKGROUND: Vascular system is affected by diseases that can seriously damage plant health by inducing browning and death of branches. This study aimed to identify and analyze the pathogenicity of Fusarium spp. isolates obtained from diseased peach branches in several peach-producing areas of China. RESULTS: We obtained and confirmed nine Fusarium isolates based on morphological and molecular characteristics. Phylogenetic relationships using a combination of rDNA-internal transcribed spacer (ITS), elongation factor (EF)-1α, and mitochondrial small subunit (mtSSU) gene sequences were analyzed. GJH-Z1, GJH-6, and GJH-1 were identified as F. avenaceum; HYR-Z3, and ZLZT-6 as F. concentricum, HH-2020-G2, and HYTZ-4 as F. solani, GG-2020-1 as F. asiaticum, SYGZ-1 as F. equiseti. Through acupuncture comparison, the pathogenicity of F. equiseti (SYGZ-1) was highest amongst nine strains. Meanwhile, F. concentricum (HYR-Z3 and ZLZT-6), and F. solaini (HYTZ-4) had a higher level of pathogenicity as revealed by impregnation. CONCLUSIONS: Our study shed light on the findings that Fusarium spp. can inflict vascular bundle browning of peach plants. Our results will extend the understanding of pathogenic diseases in China's peach industry.

Investigation of antifungal activities of myrcene on Fusarium reference strains.

Antifungal effects of myrcene, the plant-based naturel compound, were investigated on Fusarium graminearum PH-1 and Fusarium culmorum FcUK99 references, for the first time. Minimum inhibitory concentration (MIC) and half of MIC (MIC50) of both Fusarium strains against myrcene were found as 25 µg/µl and 12.5 µg/µl, respectively. MIC50 application decreased the cell viabilities in the ratios of 34.90% and 33.91% in PH-1 and FcUK99, respectively (p < 0.01). The significantly increased catalase (CAT) activity was recorded in MIC50 treated strains (p < 0.01). Apoptosis-like process and cellular oxidative stress were also monitored with acridine orange/ethidium bromide (Ao/Eb) dual staining and 2',7'-dichlorodihydrofluorescein diacetate (DCF-DA) staining. The genomic template stability (GTS) percentages were calculated as 79% for PH-1 and 71% for FcUK99 via random amplified polymorphic DNA (RAPD). Methylation polymorphism values were calculated as 53.8% and 50.6% in PH-1 and 40.4% and 39% in FcUK99 for HapII and MspI, respectively by coupled restriction enzyme digestion-random amplification (CRED-RA). Methylation-sensitive amplified polymorphism (MSAP) revealed that myrcene caused both type I and type III epigenetic modifications in both genomes. MIC50 dose caused up to 13.86 ± 0.42-fold changes in the expressions of cat, mst20, and stuA, whereas downregulation in tri5 was recorded. Myrcene application did not change the retrotransposon movement in both species by the amplifying of idiomorphic retrotransposon patterns through inter-retrotransposon polymorphism-polymerase chain reaction (IRAP-PCR). This study demonstrated that myrcene is an effective compound in the management of phytopathogenic Fusarium species by causing morphological, genetic, epigenetic, and cellular alterations, and has a potential to utilize as an antifungal agent.

Treatment of Fusarium Infection of the Central Nervous System: A Review of Past Cases to Guide Therapy for the Ongoing 2023 Outbreak in the United States and Mexico.

INTRODUCTION: Fusariosis of the central nervous system (CNS) is extremely uncommon. Treatment and outcome data from previously published cases may provide some guidance in light of the ongoing fungal meningitis outbreak in 2023 involving Fusarium spp. in the United States and Mexico. METHODS: We reviewed the published literature describing cases of invasive fusariosis of the (CNS) that included data on patient demographic characteristics, treatment, and outcome. RESULTS: Twenty-six cases met inclusion criteria. The mean age was 36 years, 55% involved females, 60% had underlying hematologic malignancy, and another 16% were on immunosuppressants. The majority of infections were from Fusarium solani species complex. Overall 72% of patients died. The majority received monotherapy with amphotericin B, although some received voriconazole monotherapy or combination therapy with amphotericin B plus voriconazole with or without adjuvant surgery. Among the survivors, 3 received amphotericin B monotherapy, 2 voriconazole monotherapy, 1 combination therapy of both, and one surgery only. CONCLUSION: The overall mortality rate in published cases of fusariosis of the CNS was high, although-unlike during the current outbreak-the preponderance of patients were severely immunocompromised. While historically the majority were treated with amphotericin B monotherapy, some recent patients were treated with voriconazole monotherapy or combination therapy with amphotericin B plus voriconazole. Current guidelines recommend monotherapy with voriconazole or lipid formulations of amphotericin B or combination of both for the treatment of invasive fusariosis, which is in line with the findings from our literature review and should be considered during the ongoing 2023 outbreak.

Ecology and diversity of culturable fungal species associated with soybean seedling diseases in the Midwestern United States.

AIMS: To isolate and characterize fungi associated with diseased soybean seedlings in Midwestern soybean production fields and to determine the influence of environmental and edaphic factors on their incidence. METHODS AND RESULTS: Seedlings were collected from fields with seedling disease history in 2012 and 2013 for fungal isolation. Environmental and edaphic data associated with each field was collected. 3036 fungal isolates were obtained and assigned to 76 species. The most abundant genera recovered were Fusarium (73%) and Trichoderma (11.2%). Other genera included Mortierella, Clonostachys, Rhizoctonia, Alternaria, Mucor, Phoma, Macrophomina and Phomopsis. Most recovered species are known soybean pathogens. However, non-pathogenic organisms were also isolated. Crop history, soil density, water source, precipitation and temperature were the main factors influencing the abundance of fungal species. CONCLUSION: Key fungal species associated with soybean seedling diseases occurring in several US production regions were characterized. This work also identified major environment and edaphic factors affecting the abundance and occurrence of these species. SIGNIFICANCE AND IMPACT OF THE STUDY: The identification and characterization of the main pathogens associated with seedling diseases across major soybean-producing areas could help manage those pathogens, and devise more effective and sustainable practices to reduce the damage they cause.

Bioactivity of mefentrifluconazole against different Fusarium spp.

Emergence and development of resistance to 14α-demethylase inhibitors (DMIs) have become a critical issue in both agriculture and medical fields. Mefentrifluconazole, the first isopropanol triazole fungicide belonging to a new subclass of DMIs, has been proposed to show high activity, minimal adverse side effects, and inconsistent cross resistance with other DMIs due to its high structural flexibility. In this study, mefentrifluconazole showed disparate inhibitory activity against the mycelium growth of seven tested Fusarium species. The most sensitive species included F. oxysporum, F. proliferatum, F. commuae, and F. fujikuroi, followed by F. equiseti and F. graminearum, while F. solani was most insensitive. Consistently, mefentrifluconazole presented the strongest inhibiting effects on conidium germination, cell membrane integrity, and ergosterol biosynthesis in F. fujikuroi, followed by F. graminearum, while F. solani ranked last. Further results indicated that all F. fujikuroi isolates causing rice bakanae disease (RBD) were sensitive to mefentrifluconazole regardless of their resistance to prochloraz, tebuconazole, carbendazim, and phenamacril. Additionally, the inoculation tests found that mefentrifluconazole presented a better protective efficacy on rice seedlings when applied 12 h before the F. fujikuroi inoculation, compared to applied 12 h post the inoculation. Overall, this study demonstrated the various bioactivity of mefentrifluconazole combating Fusarium spp. and put new insights into RBD management as well as the applications of DMIs.

Identification of Pathogenic Fusarium spp. Responsible for Root Rot of Angelica sinensis and Characterization of Their Biological Enemies in Dingxi, China.

Root rot is a serious disease in plantations of Angelica sinensis, severely reducing yield and quality and threatening sustainable production. Fusarium isolates (n = 32) were obtained from field samples of root rot tissue, leaves, and infected soil. Isolates were identified by comparison of the sequences of their internal transcribed spacer region and translation elongation factor 1-α to sequences of known species in the National Center for Biotechnology Information database. These Fusarium isolates include Fusarium tricinctum (43.75%), F. equiseti (31.25%), F. solani (9.37%), F. oxysporum (6.25%), F. acuminatum (6.25%), and F. incarnatum (3.12%). For pathogenicity testing under greenhouse conditions, seven isolates were selected based on a phylogenetic analysis, including four strains of F. tricinctum and one strain each of F. solani, F. oxysporum, and F. acuminatum. The seven isolates were all pathogenic but differed in their ability to infect: The four F. tricinctum strains were capable of causing root rot in A. sinensis at 100% incidence and were highly aggressive. Furthermore, the symptoms of root rot induced by those seven isolates were consistent with typical root rot cases in the field, but their disease severity varied. Observed histopathological preparations of F. tricinctum-infected seedlings and tissue slide results showed that this fungal species can penetrate epidermal cells and colonize the cortical cells where it induces necrosis and severe plasmolysis. Plate confrontation experiments showed that isolated rhizosphere bacteria inhibited the Fusarium pathogens that cause root rot in A. sinensis. Our results provide timely information about the use of biocontrol agents for suppression of root rot disease.

Discovery of Fusarium proliferatum f. sp. malus domestica Causing Apple Replant Disease in China.

Apple replant disease (ARD) is the most serious threat facing the apple industry globally. ARD is mainly manifested as decreased plant growth, serious root rot disease, and considerable yield loss. Microbial factors are the dominant factors leading to the occurrence of ARD. Research on soil-borne pathogenic fungi leading to the occurrence of ARD in China is limited. In the present study, we selected 16 replanting orchards from the Northwest Loess region and around the Bohai Gulf. Diseased roots and rhizosphere soil from healthy apple trees and trees showing ARD symptoms were sampled at random. High-throughput sequencing was used to study the fungal communities in the rhizosphere soil, which showed that the composition of the rhizosphere soil fungal community of ARD-symptomatic and healthy apple trees was different. Nectriaceae at the family level and Fusarium at the genus level dominated the rhizosphere soil fungal community in the two regions, while for healthy apple trees, the relative abundance of Mortierella, Minimedusa, Tetracladium, and Chaetomium was higher. Tissue separation and serial dilution were used to separate fungi, and a total of 89 genera and 219 species were obtained, most of which were Fusarium. Fusarium was further confirmed to be the most abundant pathogen species leading to the occurrence of ARD in China through pathogenicity assays. A pathogenicity assay was carried out by the dip-and-cut technique using different host plants. It was found that Fusarium MR5 showed strong aggressiveness to apple rootstocks. Diseased seedlings specifically exhibited chlorosis of the leaves, browning from the edge of the leaf, followed by rolling and yellowing of the leaves, resulting in wilting and eventually death. Strain MR5 was preliminarily identified as F. proliferatum according to the morphological and cultural characteristics. A maximum likelihood analysis of identities based on six gene sequence (ITS, TUB2, IGS, mtSSU, RPB2, and the TEF gene) alignments between the MR5 strain and other strains showed 99 to 100% homology with F. proliferatum. Based on our test results, strain MR5 was identified as F. proliferatum f. sp. malus domestica, which is of great significance for finding new measures to control ARD in China.

Identification, Pathogenicity, and Genetic Diversity of Fusarium spp. Associated with Maize Sheath Rot in Heilongjiang Province, China.

Maize sheath rot is a prevalent maize disease in China. From 2020 to 2021, symptomatic samples were collected from the main maize-growing regions of Heilongjiang province. To clarify the population and genetic diversity, as well as the virulence of pathogens responsible for maize sheath rot, a total of 132 Fusarium isolates were obtained and used for follow-up studies. Ten Fusarium species were identified based on morphological characteristics, and phylogenetic analysis was conducted using the TEF-1α gene sequences, including F. verticillioides (50.00%), F. subglutinans (18.94%), the Fusarium incarnatum-equiseti species complex (14.39%), F. temperatum (5.30%), F. acuminatum (3.03%), F. solani (2.27%), F. sporotrichioides (2.27%), F. tricinctum (1.52%), F. asiaticum (1.52%), and F. proliferatum (0.76%). All 10 Fusarium species could produce oval-to-annular lesions on maize sheath, and the lesions were grayish yellow to dark brown in the center and surrounded by a dark gray-to-dark brown halo. Of these, F. tricinctum and F. proliferatum showed significantly higher virulence than the other Fusarium species. In addition, haplotype analysis based on the concatenated sequences of the ITS and TEF-1a genes showed that 99 Fusarium isolates which belonged to the Fusarium fujikuroi species complex-consisting of F. verticillioides isolates, F. subglutinans isolates, F. temperatum isolates, and F. proliferatum isolates-could be grouped into 10 haplotypes, including 5 shared haplotypes (Haps 1, 2, 4, 5, and 6) and 5 private haplotypes (Haps 3, 7, 8, 9, and 10). Furthermore, the F. verticillioides clade in the haplotype network was radial with the center of Hap 2, suggesting that population expansion occurred. This research showed that Fusarium species associated with maize sheath rot in Heilongjiang province are more diverse than previously reported, and this is the first time that F. subglutinans, F. temperatum, F. solani, F. sporotrichioides, F. tricinctum, and F. acuminatum have been confirmed as the causal agents of maize sheath rot in Heilongjiang province.

Trichoderma versus Fusarium-Inhibition of Pathogen Growth and Mycotoxin Biosynthesis.

This study evaluated the ability of selected strains of Trichoderma viride, T. viridescens, and T. atroviride to inhibit mycelium growth and the biosynthesis of mycotoxins deoxynivalenol (DON), nivalenol (NIV), zearalenone (ZEN), α-(α-ZOL) and ß-zearalenol (ß-ZOL) by selected strains of Fusarium culmorum and F. cerealis. For this purpose, an in vitro experiment was carried out on solid substrates (PDA and rice). After 5 days of co-culture, it was found that all Trichoderma strains used in the experiment significantly inhibited the growth of Fusarium mycelium. Qualitative assessment of pathogen-antagonist interactions showed that Trichoderma colonized 75% to 100% of the medium surface (depending on the species and strain of the antagonist and the pathogen) and was also able to grow over the mycelium of the pathogen and sporulate. The rate of inhibition of Fusarium mycelium growth by Trichoderma ranged from approximately 24% to 66%. When Fusarium and Trichoderma were co-cultured on rice, Trichoderma strains were found to inhibit DON biosynthesis by about 73% to 98%, NIV by about 87% to 100%, and ZEN by about 12% to 100%, depending on the pathogen and antagonist strain. A glycosylated form of DON was detected in the co-culture of F. culmorum and Trichoderma, whereas it was absent in cultures of the pathogen alone, thus suggesting that Trichoderma is able to glycosylate DON. The results also suggest that a strain of T. viride is able to convert ZEN into its hydroxylated derivative, ß-ZOL.