Plant Pathogenic Fungi Research Articles

Regev, a New Hybrid Fungicide for Control of Cercospora Leaf Spot on Sugar Beet

Cercospora leaf spot (CLS) is one of the most yield-limiting foliar disease of sugar beet in the United States. Standard practices to control this disease include selection of tolerant varieties, crop rotations, and reliance on fungicides. However, genetic resistance still requires some fungicides for acceptable control under high disease pressure, and there are only a limited number of fungicides registered for control of this disease. Regev® is a newly registered hybrid fungicide for sugar beet disease control that contains eight biological active ingredients from tea tree oil (TTO) and one active ingredient from the synthetic fungicide, difenoconazole. Tea tree oil provides a new and unique mode of activity against both fungal and bacterial plant pathogens, and it has a low resistance risk. Eight trials were conducted to evaluate Regev for efficacy against Cercospora leafspot, and sugar beet yield and quality benefits in Minnesota and Michigan. Five of the trials had slightly different treatments with the primary goal of allowing trialists to conduct trials with treatments that were appropriate for their locations. However, one of the trials at the Northwest Research and Outreach Center (NWROC) of the University of Minnesota in Crookston, Minnesota, was conducted three times over the years 2021, 2022, and 2023. Across the eight trials, spray programs that included Regev provided control of Cercospora leaf spot equal to standard fungicide programs.

Design, synthesis, and biological activities of arecoline derivatives containing 1,3,4-oxadiazole structure

Pesticides play an important role in the development of agriculture, as they can prevent and control crop diseases and pests, improve crop yield and quality. However, the abuse and improper use of pesticides can lead to negative impacts such as environmental pollution and pest resistance issues. There is an urgent need to develop green, safe, and efficient pesticides. In this work, natural product arecoline was selected as parent structure, a series of arecoline derivatives were designed, synthesized, and systematically investigated antiviral activities against tobacco mosaic virus (TMV). These compounds were found to have good to excellent anti-TMV activities for the first time. The antiviral activities of 4a, 4 h, 4 l, 4p, 6a, 6c, and 6f are higher than that of ningnanmycin. Compounds 4 h (EC50 value 146 µg/mL) and 4p (EC50 value 161 µg/mL) with simple structures and excellent activities emerged as new antiviral candidates. We chose 4 h to further investigate the antiviral mechanism, which revealed that it can cause virus fragmentation by acting on the viral coat protein (CP). We further validated this result through molecular docking. These compounds also displayed broad-spectrum fungicidal activities against 8 plant pathogenic fungi. This work lays the theoretical foundation for the application of arecoline derivatives in the agricultural field.

Antifungal properties and molecular docking of ZnO NPs mediated using medicinal plant extracts

Significant postharvest losses and food safety issues persist in many developing nations, primarily due to fungal activities, including mycotoxin production. In this study, green synthesised zinc oxide nanoparticles (ZnO-NPs) were prepared from leaf extracts of Syzygium cordatum (ZnO 1), Lippia javanica (ZnO 2), Bidens pilosa (ZnO 3), and Ximenia caffra (ZnO 4). Physicochemical characteristics of the ZnO-NPs were determined using X-ray diffraction (XRD), Fourier transmission Infrared spectroscopy and ultraviolet–visible (UV–vis) spectroscopy, scanning electron microscopy (SEM), and transmission electron microscopy (TEM). The XRD analysis confirmed the presence of a wurtzite crystal structure in the hexagonal shape of the ZnO nanoparticles (NPs), with an average size ranging between 25 and 43 nm. The microscopic examination of the morphology revealed the presence of spherical particles with sizes ranging from 37 to 47 nm in diameter. The antifungal efficacy of the ZnO-NPs was assessed against pathogenic plant fungi, including Botrytis sp. (STEU 7866), Penicillium sp. (STEU 7865), and Pilidiella granati (STEU 7864), using the poisoned food technique. Further antifungal evaluation of the ZnOPs was performed using the broth microdilution assay. A significant interaction between the type of ZnO-NPs and fungal species was observed, with the highest susceptibility in Mucor sp. to ZnO 2, achieving over 50% inhibition. Penicillium sp. also showed high susceptibility to all ZnO-NPs. Molecular docking results confirmed the strong H-bonding interactions of ZnO-NPs with fungal receptors in Mucor sp. and Penicillium sp., Botrytis sp. and P. granati exhibited the least susceptibility. Further tests revealed that ZnO 2 exhibited the highest inhibitory effect on Botrytis sp., with a low minimum inhibitory concentration (MIC) of 25 µg/mL, attributed to its larger positive zeta potential. This study indicates that ZnO NPs, particularly those mediated using Lippia javanica (ZnO 2), have promising potential as effective antifungal agents, which could play a significant role in reducing postharvest decay and losses.

Re-routing MAP kinase signaling for penetration peg formation in predator yeasts.

Saccharomycopsis yeasts are natural organic sulfur auxotrophs due to lack of genes required for the uptake and assimilation of sulfate/sulfite. Starvation for methionine induces a shift to a predatory, mycoparasitic life strategy that is unique amongst ascomycetous yeasts. Similar to fungal plant pathogens that separated from Saccharomycopsis more than 400 million years ago, a specialized infection structure called penetration peg is used for prey cell invasion. Penetration pegs are highly enriched with chitin. Here we demonstrate that an ancient and conserved MAP kinase signaling pathway regulates penetration peg formation and successful predation in the predator yeast S. schoenii. Deletion of the MAP kinase gene SsKIL1, a homolog of the Saccharomyces cerevisiae ScKSS1/ScFUS3 and the rice blast Magnaporthe oryzae MoPMK1 genes, as well as deletion of the transcription factor SsSTE12 generate non-pathogenic mutants that fail to form penetration pegs. Comparative global transcriptome analyses using RNAseq indicate loss of the SsKil1-SsSte12-dependent predation response in the mutant strains, while a methionine starvation response is still executed. Within the promoter sequences of genes upregulated during predation we identified a cis-regulatory element similar to the ScSte12 pheromone response element. Our results indicate that, re-routing MAP-kinase signaling by re-wiring Ste12 transcriptional control towards predation specific genes contributed to the parallel evolution of this predacious behaviour in predator yeasts. Consequently, we found that SsSTE12 is dispensable for mating.

Distribution and Sequestration of Cercosporin by Cercospora cf. flagellaris.

Plant-pathogenic fungi produce toxins as virulence factors in many plant diseases. In Cercospora leaf blight of soybean caused by Cercospora cf. flagellaris, symptoms are a consequence of the production of a perylenequinone toxin, cercosporin, which is light-activated to produce damaging reactive oxygen species. Cercosporin is universally toxic to cells, except to the cells of the producer. The current model of self-resistance to cercosporin is largely attributed to the maintenance of cercosporin in a chemically reduced state inside hyphae, unassociated with cellular organelles. However, in another perylenequinone-producing fungus, Phaeosphaeria sp., the toxin was specifically sequestered inside lipid droplets (LDs) to prevent reactive oxygen species production. This study hypothesized that LD-based sequestration of cercosporin occurred in C. cf. flagellaris and that lipid-inhibiting fungicides could inhibit toxin production. Confocal microscopy using light-cultured C. cf. flagellaris indicated that 3-day-old hyphae contained two forms of cercosporin distributed in two types of hyphae. Reduced cercosporin was uniformly distributed in the cytoplasm of thick, primary hyphae, and, contrary to previous studies, active cercosporin was observed specifically in the LDs of thin, secondary hyphae. The production of hyphae of two different thicknesses, a characteristic of hemibiotrophic plant pathogens, has not been documented in C. cf. flagellaris. No correlation was observed between cercosporin production and total lipid extracted, and two lipid-inhibiting fungicides had little effect on fungal growth in growth-inhibition assays. This study lays a foundation for exploring the importance of pathogen lifestyle, toxin production, and LD content in the pathogenicity and symptomology of Cercospora.

An assessment of twenty-three mycorrhizal inoculants reveals limited viability of AM fungi, pathogen contamination, and negative microbial effect on crop growth for commercial products

Ensuring sustainable agriculture is crucial amidst global challenges, demanding effective methods to enhance soil health and nutrient cycling. Microbial inoculants, particularly arbuscular mycorrhizal (AM) fungi, offer promising solutions. However, concerns persist regarding the efficacy and quality control of commercial products. Past work assessing commercial inoculants have not controlled for fertilizers added to individual products when assessing product effects under typical use.This study examines twenty-three mycorrhizal inoculants using conventions of organic production to shed light on differences between laboratory grown fungi, commercial products, and field soil. Employing a comprehensive approach, inoculants were assessed through spore enumeration, root infection potential, and crop growth response.The results uncover significant shortcomings in many commercial products compared to laboratory grown fungi. Key findings include discrepancies of up to 100 % in reported propagule counts versus spore concentrations, insufficient root colonization by commercial inoculants, and contamination by fungal plant pathogens, particularly Olpidium, in products. Moreover, while laboratory grown fungi exhibited superior symbiotic relationships with host plants due to increased colonization abilities and crop benefit, commercial inoculants often failed to deliver significant growth benefits when fertilizers are controlled for.These findings highlight the urgent need for improved standards and practices within the commercial inoculant industry.

Botrytis cinerea combines four molecular strategies to tolerate membrane-permeating plant compounds and to increase virulence

Saponins are plant secondary metabolites comprising glycosylated triterpenoids, steroids or steroidal alkaloids with a broad spectrum of toxicity to microbial pathogens and pest organisms that contribute to basal plant defense to biotic attack. Secretion of glycosyl hydrolases that enzymatically convert saponins into less toxic products was thus far the only mechanism reported to enable fungal pathogens to colonize their saponin-containing host plant(s). We studied the mechanisms that the fungus Botrytis cinerea utilizes to be tolerant to well-characterized, structurally related saponins from tomato and Digitalis purpurea. By gene expression studies, comparative genomics, enzyme assays and testing a large panel of fungal (knockout and complemented) mutants, we unraveled four distinct cellular mechanisms that participate in the mitigation of the toxic activity of these saponins and in virulence on saponin-producing host plants. The enzymatic deglycosylation that we identified is novel and unique to this fungus-saponin combination. The other three tolerance mechanisms operate in the fungal membrane and are mediated by protein families that are widely distributed in the fungal kingdom. We present a spatial and temporal model on how these mechanisms jointly confer tolerance to saponins and discuss the repercussions of these findings for other plant pathogenic fungi, as well as human pathogens.

Regional diversity and leaf microbiome interactions of the fungal maize pathogen Exserohilum turcicum in Switzerland: A metagenomic analysis.

The spread and adaptation of fungal plant pathogens in agroecosystems are facilitated by environmental homogeneity. Metagenomic sequencing of infected tissues allowed us to monitor eco-evolutionary dynamics and interactions between host, pathogen and plant microbiome. Exserohilum turcicum, the causal agent of northern corn leaf blight (NCLB) in maize, is distributed in multiple clonal lineages throughout Europe. To characterize regional pathogen diversity, we conducted metagenomic DNA sequencing on 241 infected leaf samples from the highly susceptible Swiss maize landrace Rheintaler Ribelmais, collected over 3 years (2016-2018) from an average of 14 agricultural farms within the Swiss Rhine Valley. All major European clonal lineages of E. turcicum were identified. Lineages differ by their mating types which indicates potential for sexual recombination and rapid evolution of new pathogen strains, although we found no evidence of recent recombination. The associated eukaryotic and prokaryotic leaf microbiome exhibited variation in taxonomic diversity between years and locations and is likely influenced by local weather conditions. A network analysis revealed distinct clusters of eukaryotic and prokaryotic taxa that correlates with the frequency of E. turcicum sequencing reads, suggesting causal interactions. Notably, the yeast genus Metschnikowia exhibited a strongly negative association with E. turcicum, supporting its known potential as biological control agent against fungal pathogens. Our findings show that metagenomic sequencing is a useful tool for analysing the role of environmental factors and potential pathogen-microbiome interactions in shaping pathogen dynamics and evolution, suggesting their potential for effective pathogen management strategies.

Glyoxal oxidase-mediated detoxification of reactive carbonyl species contributes to virulence, stress tolerance, and development in a pathogenic fungus.

Reactive carbonyl and oxygen species (RCS/ROS), often generated as metabolic byproducts, particularly under conditions of pathology, can cause direct damage to proteins, lipids, and nucleic acids. Glyoxal oxidases (Gloxs) oxidize aldehydes to carboxylic acids, generating hydrogen peroxide (H2O2). Although best characterized for their roles in lignin degradation, Glox in plant fungal pathogens are known to contribute to virulence, however, the mechanism underlying such effects are unclear. Here, we show that Glox in the insect pathogenic fungus, Metarhizium acridum, is highly expressed in mycelia and during formation of infection structures (appressoria), with the enzyme localizing to the cell membrane. MaGlox targeted gene disruption mutants showed RCS and ROS accumulation, resulting in cell toxicity, induction of apoptosis and increased autophagy, inhibiting normal fungal growth and development. The ability of the MaGlox mutant to scavenge RCS was significantly reduced, and the mutant exhibited increased susceptibility to aldehydes, oxidative and cell wall perturbing agents but not toward osmotic stress, with altered cell wall contents. The ΔMaGlox mutant was impaired in its ability to penetrate the host cuticle and evade host immune defense resulting in attenuated pathogenicity. Overexpression of MaGlox promoted fungal growth and conidial germination, increased tolerance to H2O2, but had little to other phenotypic effects. Transcriptomic analyses revealed downregulation of genes related to cell wall synthesis, conidiation, stress tolerance, and host cuticle penetration in the ΔMaGlox mutant. These findings demonstrate that MaGlox-mediated scavenging of RCS is required for virulence, and contributes to normal fungal growth and development, stress resistance.

Soil Fungal Pathogens in Pinus pinaster Mature Reforestation: Silvicultural Treatments Effects.

Soil fungal communities play a key role in multiple functions and ecosystem services within forest ecosystems. Today, forest ecosystems are subject to multiple environmental and anthropogenic disturbances (e.g., fire or forest management) that mainly lead to changes in vegetation as well as in plant-soil interactions. Soil pathogens play an important role in controlling plant diversity, ecosystem functions, and human and animal health. In this work we analyzed the response of soil plant pathogenic fungi to forest management in a Pinus pinaster reforestation. We started from an experimental design, in which forest thinning and gap cutting treatments were applied at different intensities and sizes, respectively. The fungal communities of plant pathogens in spring were described, and the effect of the silvicultural treatments was evaluated 5 years after their application, as were the possible relationships between soil plant pathogenic fungal communities and other environmental factors. Only a strong low thinning treatment (35% basal area) was able to generate homogeneous changes in soil pathogenic diversity. In the gaps, only the central position showed significant changes in the soil plant pathogenic fungi community.

Inactivation of Fusarium verticilliordes by dielectric barrier discharge plasma and its mechanism

Fusarium is a kind plant pathogenic fungus, resulting in severe crops infection and food safety issues. The development of novel strategy to inactive Fusarium is in urgent need. In this study, the inactivation effect and mechanism of dielectric barrier discharge (DBD) plasma on Fusarium verticillioides was studied. The results showed that the fungicidal effect of DBD plasma was positively correlated with the voltage strength and treatment time, which could reach 100% sterilizing rate at 60–80 V. The scanning electron microscope (SEM) observation indicated the integrity of the cell wall and cell membrane of the treated F. verticilliordes was damaged, leading to leakage of the contents. The changes of mitochondrial membrane potential, adenosine 5′-triphosphate (ATP) content, cell viability and reactive substances (intracellular reactive oxygen species (ROS), intracellular hydrogen peroxide (H2O2), ozone (O3) and reactive nitrogen species (RNS) (NO2−, NO3−, etc.)) were also tested to better understand the fungicidal mechanism of cold plasma. These results demonstrated DBD plasma could be a promising technique for Fusarium inactivation in food.

A nanoscale γ-cyclodextrin metal-organic framework loaded with eugenol (CMFE) for controlling F. graminearum on the wheat during harvest time

The loss of grains in the production chain caused huge waste worldwide, and the control of plant pathogenic fungi during harvest time is a critical solution to mitigate the damage. Eugenol, an important component of clove oil (Syzygium aromaticum L.), has antioxidant, antifungal, and other activities. In this study, the unstable eugenol was incorporated into the nanoscale γ-cyclodextrin metal-organic framework (γ-CD-MOF), a cubic porous structure synthesized by vapor diffusion method. The investigation of morphologies, functional groups, crystalline structures and weight losing behaviors demonstrated the inclusion complex prepared from γ-CD-MOF and eugenol (CMFE). Compared with the γ-CD loaded with eugenol (CE), CMFE exhibited an improved loading rate with 4.06 % (w/w). And CMFE enhanced the stability of its inner eugenol against degradation by light and air. Moreover, the CMFE demonstrated a significant anti-F. graminearum activity both in the medium and on the wheat grains. The inhibition rate reached 83.8 % at 8 mg/mL in vitro, while the growth of the hyphae was effectively restrained at 4 mg/g in vivo. CMFE could damage the mycelial cell wall and trigger intracellular reactive oxygen species (ROS) accumulation, thus exhibiting its anti-F. graminearum activity. Our results showed that the biocompatible CMFE would be an environment-friendly substitute to chemical fungicides for the prevention of fungal contamination caused by F. graminearum.

Warming and rainfall reduction alter soil microbial diversity and co-occurrence networks and enhance pathogenic fungi in dryland soils

In this 9-year manipulative field experiment, we examined the impacts of experimental warming (2 °C, W), rainfall reduction (30 % decrease in annual rainfall, RR), and their combination (W + RR) on soil microbial communities and native vegetation in a semi-arid shrubland in south-eastern Spain. Warming had strong negative effects on plant performance across five coexisting native shrub species, consistently reducing their aboveground biomass growth and long-term survival. The impacts of rainfall reduction on plant growth and survival were species-specific and more variable. Warming strongly altered the soil microbial community alpha-diversity and changed the co-occurrence network structure. The relative abundance of symbiotic arbuscular mycorrhizal fungi (AMF) increased under W and W + RR, which could help buffer the direct negative impacts of climate change on their host plants nutrition and enhance their resistance to heat and drought stress. Indicator microbial taxa analyses evidenced that the marked sequence abundance of many plant pathogenic fungi, such as Phaeoacremonium, Cyberlindnera, Acremonium, Occultifur, Neodevriesia and Stagonosporopsis, increased significantly in the W and W + RR treatments. Moreover, the relative abundance of fungal animal pathogens and mycoparasites in soil also increased significantly under climate warming. Our findings indicate that warmer and drier conditions sustained over several years can alter the soil microbial community structure, composition, and network topology. The projected warmer and drier climate favours pathogenic fungi, which could offset the benefits of increased AMF abundance under warming and further aggravate the severe detrimental impacts of increased abiotic stress on native vegetation performance and ecosystem services in drylands.

Inhibition of Botrytis cinerea and Escherichia coli by Lactic Acid Bacteria on Leafy Vegetables

The evaluation of the potentiality of lactic acid bacteria (LAB) strains isolated from fermented products to inhibit Botrytis cinerea and Escherichia coli O157:H7 growth on spinach and lettuce was conducted. From a total of forty LAB strains tested, three were selected due to their high inhibitory effect on plant pathogenic fungi. The identification of these isolates based on a 16S rRNA gene fragment sequence analysis confirmed the genus of Levilactobacillus sp. and Lactiplantibacillus sp. An effective method of coating LAB isolates on the lettuce and spinach surface was developed. The leaves were immersed in bacterial suspension (5.0 × 106 cfu mL−1) for 4 s and drained on tissue paper. LAB survived on lettuce and spinach leaves for 8 days at 6 log10 cfu g−1. Additionally, these bacteria decreased the number of filamentous fungi on the leaves. These isolates were found to inhibit the growth of B. cinerea and E. coli O157:H7 in vitro conditions in growing microbiological media. Their efficacy was confirmed in vivo conditions. These isolates inhibited the development of grey mould caused by B. cinerea on lettuce leaves. Two LAB isolates reduced the abundance of the pathogenic bacterium E. coli on spinach leaves by about 0.7 log10 cfu g−1. In glasshouse conditions, LAB stimulated the growth of examined plants. The lactic acid bacteria used in this study showed the capacity to be used as possible alternatives to chemical compounds in the protection of leafy vegetables against grey mould and for a decrease in E. coli O157:H7 contamination.

GC-MS analysis and antifungal potential of flower extract of Acacia nilotica subsp. Indica against Macrophomina phaseolina

Macrophomina phaseolina is a wide host ranged soil-borne fungal plant pathogen. It infects more than 500 host plant species belonging to 100 families. Many important oil-seed and leguminous crops are known to be attacked by this devastating plant pathogen. In the present study, antifungal potential of flowers of a leguminous tree Acacia nilotica subsp. indica, was assessed against this pathogen through bioassays guided fractionation. Initially, methanolic extracts of 1 %–5 % of leaf, flower, root-bark and stem-bark of the plant species under consideration were evaluated for their antifungal potential against the target pathogen. Among these, the best antifungal activity was shown by flower extract. The reduction in growth of the test fungal strain was 27–49 %, 4–40 % and 2–27 % due to flower, root-bark and leaf extracts, respectivey, over control. Flower extract was partitioned using n-hexane, chloroform, ethyl acetate and n-butanol as the solvents. Bioassays guided study of these fractions of methanolic extract of flower revealed that high antifungal potential was shown by n-hexane and chloroform fractions against M. phaseolina causing 26–53 % and 28–50 % decline in fungal biomass, respectively, as compared to that of control. GC-MS analysis of chloroform fraction revealed the presence of 27 compounds in this fraction. Among these cyclopentanol,-1-methyl (10.93 %) was the predominant compound followed by methyl, 4,4-dimethyl butanoate (7.04 %), 1-pentanol (6.80 %), 2-propanol, 1-cyclopropyl (6.11 %), 1H,imidazole-4-5-dihydro-2-methyl (5.93 %), trichloroethane (5.91 %), carbonic acid-ethyl hexyl ester (4.59 %), 1,4-butandiol,2,3-bis(methylene)- (4.54 %) and [S]-3,4-dimethyl pentanol (4.48 %).

Plant Disease: A Growing Threat to Global Food Security

The escalating global population has led to an increased demand for both quantity and quality in food production. Throughout history, plant diseases have posed significant threats to agricultural output by causing substantial food losses annually while also compromising product quality. Accurate identification of pathogens, clarifying the pathogenic mechanism of pathogens, and understanding the interaction between pathogens and hosts are important for the control of plant diseases. This Special Issue, “Research Progress on Pathogenicity of Fungi in Crops”, belongs to the section “Pest and Disease Management” of Agronomy. It contains research papers on the identification and phylogeny of fungal pathogens, the molecular genetics of plant fungal pathogens, the molecular mechanisms of fungal pathogenicity, and the molecular basis of the interaction between fungi and crops. These studies encapsulate efforts to understand disease systems within current genomics, transcriptomics, proteomics, and metabolomics studies, highlighting research findings that could be future targets for crop disease and pest control. The studies presented in this Special Issue promote the progress of fungal pathogenicity research in crops and provide a scientific basis for future disease control, which is of great significance for sustainable agricultural development and global food security.

Microscopic and Transcriptomic Analyses to Elucidate Antifungal Mechanisms of Bacillus velezensis TCS001 Lipopeptides against Botrytis cinerea.

Botrytis cinerea is an important fungal pathogen that causes gray mold disease in plants. Previously, Bacillus velezensis TCS001 live culture presented broad-spectrum antifungal activity against various plant pathogenic fungi and oomycetes, particularly B. cinerea. Here, the bioactivity of lipopeptides produced by TCS001 against B. cinerea was investigated. The IC50 values of the crude lipopeptide extract (CLE) from TCS001 to suppress mycelial growth and conidial germination were 14.20 and 49.39 mg/L, respectively. SEM and TEM imaging revealed that CLE caused morphological deformities and ultrastructural changes in the mycelium. Transcriptomic analyses combined with ΔBcpsd mutant construction demonstrated that the CLE could confer antifungal activity via suppressing Bcpsd expression in the pathogen. In addition, the CLE activated the plant immune system by increasing the content of defense-related enzymes and the expression of marker genes in immunity signaling pathways in cucumber plants. Therefore, TCS001 CLE could be potentially developed into biopesticides for the biocontrol of gray mold disease.

Magnaporthe-Unique Gene MUG1 Is Important for Fungal Appressorial Penetration, Invasive Hyphal Extension, and Virulence in Rice Blast Fungi.

Species-unique genes that encode specific proteins and have no homologs in other species play certain roles in the evolution of species and adaptations to external environments. Nevertheless, the biological roles of unique genes in plant pathogenic fungi remain largely unknown. Here, four Magnaporthe-unique genes (MUG1-MUG4), which were highly expressed during the early infection stages, were functionally characterized in the rice blast fungus Magnaporthe oryzae. Subcellular localization assays revealed that Mug1, Mug2, and Mug4 were localized to the cytoplasm and that Mug3 was localized into the nuclei. Furthermore, through gene knockout and phenotypic analysis, only MUG1 was found to be indispensable for fungal virulence and conidiation. Detailed microscopic analysis revealed that the deletion mutants of MUG1 clearly exhibited reduced appressorial turgor pressure and invasive hyphal development. Taken together, our findings indicate that the Magnaporthe-unique gene MUG1 plays a vital role in infection-related morphogenesis and virulence in rice blast fungi and suggest the specific and important roles of species-unique genes.

A pooled mycoviral resource in a strain of Rhizoctonia solani are regulators of fungal virulence

Rhizoctonia solani is a widespread and devastating soil-borne plant fungal pathogen that causes diseases, including rice sheath blight, which are difficult to control. Some mycoviruses are potential biocontrol agents for the control of fungal diseases. In order to investigate the factors that influence the virulence of R. solani and search for mycoviruses with the potential for biocontrol of R. solani, a rice-infecting R. solani strain, ZJXD1–1, was isolated and confirmed to contain eight mycoviruses via dsRNA extraction and high-throughput sequencing. The identified mycoviruses belong to families of Endornaviridae (RsEV11 and RsEV12) and Mitoviridae (RsMV125 to RsMV129), and an unclassified Toti-like clade (RsTLV1). The C39 domain in RsEV12, which shares a close evolutionary relationship with bacteria, is observed for the first time in a mycovirus. Strains with different virus combinations were obtained through viral horizontal transfer, and pathogenicity test deduced that the Endornaviruses RsEV11 and RsEV12, and Mitovirus RsMV129 might potentially enhance the pathogenicity of R. solani, while RsMV125 might reduce the virulence or interfere with the function of other Mitoviruses. Furthermore, virus curing via protoplast regeneration and viral horizontal transfer demonstrated that RsMV129 is the causal agent of R. solani hypervirulence. Overall, our study provided the resource pool of viruses that may contribute to the discovery of new biocontrol agents against R. solani and enhance our understanding of the pathogenesis of R. solani regulated by mycoviruses.

Synthesis, application and molecular docking of modified cellulose with diaminoguanidine as complexing agent for selective separation of Cu (II), Cd (II) and Hg (II) ions from alum sample

A new modified cellulose with diaminoguanidine (Cel-Gua) synthesized for specific recovery of Cu (II), Cd (II), and Hg (II) from the alum sample. Cellulose was silanized by 3-chloropropyltrimethoxysilane and then was modified with diaminoguanidine to obtain N-donor chelating fibers. Fourier transform-infrared spectroscopy, scanning electron microscopy, X-ray diffraction, zeta potential, electrons disperse X-ray analysis, elemental analyses (C, H and N), and thermogravimetric analysis were used for characterization. Factors influencing the adsorption were thoroughly examined. Under the optimal conditions, the Cel-Gua sorbent displayed maximum adsorption capacities of 94.33, 112.10 and 95.78 mg/g for Cu (II), Cd (II), and Hg (II), respectively. The sorption process of metal ions is equipped by kinetic model PSO and Langmuir adsorption isotherm. The calculated thermodynamic variables confirmed that the adsorption of Cu (II), Cd (II) and Hg (II) by Cel-Gua sorbent is a spontaneous and exothermic process. In our study, we used the molecular operating environment software to conduct molecular docking simulations on the Cel-Gua compound. The results of the docking simulations showed that the Cel-Gua compound displayed greater potency and a stronger affinity for the Avr2 effector protein derived from Fusarium oxysporum, a fungal plant pathogen (code 5OD4). The adsorbent was stable for 7 cycles, thus allowing its safe reutilization.