Search Results (182)

Entomopathogenic fungi are increasingly recognized as multifunctional bioinputs, but robust methodological approaches are needed to evaluate their compatibility with established microbial inoculants and their effects on plant performance. Our study proposes an integrative framework to assess Metarhizium–Bradyrhizobium interactions in soybean (Glycine max), combining laboratory compatibility screening with greenhouse assessment. First, Metarhizium anisopliae LCM S04 and Metarhizium brunneum LCM S11 were tested against Bradyrhizobium diazoefficiens BR 85 (=SEMIA 5080) and Bradyrhizobium japonicum BR 86 (=SEMIA 5079) using dual-culture assays in a medium that supported the growth of both fungal and bacterial partners, allowing direct evaluation of intermicrobial compatibility. Subsequently, the microorganisms were evaluated in soybean under greenhouse conditions through direct seed inoculation. Each seed received bacterial culture, fungal suspension, or both (including absolute and nitrogen-fertilized controls). Plants were maintained under controlled conditions for 47 days. Plant height, leaf production, branching, nodulation, biomass, and root morphology were assessed. No inhibition zones were observed in vitro, and co-inoculation did not impair nodulation. The combination BR 86 + LCM S04 improved fine root number in soybean. This framework provides a reproducible approach for evaluating microbial compatibility and functional bioinput benefits and can be adapted to crops other than soybean. Full article

This study aims to elucidate the soil fungal community structure, the distribution of entomopathogenic fungi (EPF) resources, and their driving mechanisms in the alpine grassland of the northern side of the eastern Tanggula Mountains in the Qinghai–Tibet Plateau. Five representative sites were established along an elevational gradient of 4000–5100 m, and 15 soil samples were collected. High-throughput sequencing (ITS region) and an insect baiting method were combined to analyze fungal community composition and diversity. LEfSe analysis, random forest modeling, redundancy analysis (RDA), and network analysis were employed to reveal the environmental driving mechanisms of community assembly. A total of 412 fungal species belonging to 303 genera were annotated, with Ascomycota as the dominant phylum. At the high-elevation site Jie-duo Town, Zaduo County (ZD-JD), the cold-tolerant ectomycorrhizal fungus Inocybe was absolutely dominant (17.18%). The low-elevation site Dong-ba Town, Nangqian County (NQ-DB), was enriched with the saprotrophic genus Mortierella (10.34%) and EPF taxa. The mid-elevation site Zhuo-xiao Town, Nangqian County (NQ-ZX), was characterized by Solicocozyma (11.88%) and exhibited the highest network complexity. Alpha diversity decreased with increasing elevation. A total of 35 EPF strains were isolated via the baiting method, predominantly Metarhizium anisopliae, M. brunneum, and M. robertsii, with the highest number of isolates (16 strains) recovered from NQ-DB. Elevation indirectly drove community assembly by regulating temperature and plant productivity. The soil fungal community structure in the alpine grassland of the eastern Tanggula Mountains exhibits significant spatial heterogeneity along the elevational gradient. Low-elevation habitats serve as hotspots for EPF resources, while high-elevation isolates harbor potential stress-resistance genes. The combination of culture-dependent and sequencing-based approaches provides a reliable strategy for microbial resource surveys in extreme environments. Full article

The appressorium is a specialized infection structure formed by Metarhizium anisopliae during host invasion. To investigate the correlation between appressorium formation and fungal pathogenicity, as well as its impact on insect cuticular metabolism, different concentrations of sulforaphane were used to inhibit the appressorium formation rate of M. anisopliae. The relationship between appressorium formation rate and pathogenicity against Opisina arenosella larvae was evaluated, and LC-MS-based metabolomics was employed to characterize changes in cuticular compounds during the appressorium formation stage, thereby elucidating the chemical responses of the insect cuticle to appressorium formation. The appressorium formation rate of M. anisopliae was significantly and positively correlated with its pathogenicity (p ≤ 0.05). As the appressorium formation rate increased, pathogenicity against O. arenosella larvae increased and the killing speed accelerated. LC-MS metabolomics revealed that after appressorium formation, 102 differential cuticular compounds unique to O. arenosella larvae were identified, mainly including benzenes and substituted derivatives, amino acids and derivatives, and heterocyclic compounds. In addition, metabolic pathways associated with immune defense (tyrosine metabolism), antifungal defense (histidine metabolism), and toxin degradation (flavonoid degradation) in the larval cuticle were activated. These results demonstrate that the appressorium plays an important role in host infection by M. anisopliae, markedly alters cuticular metabolism, and activates defense- and detoxification-related metabolic pathways in the host. This study provides a theoretical basis for further investigations into fungus–insect cuticle interactions. Full article

Subterranean termites, particularly Odontotermes obesus, cause severe damage to forest nurseries and plantations in arid and semi-arid ecosystems. This study demonstrates the dual functional role of endophytic entomopathogenic fungi, Metarhizium anisopliae and Beauveria bassiana, in termite suppression and induction of plant defense responses. Laboratory bioassays revealed significantly higher virulence of M. anisopliae, with a lower LT₅₀ (lethal time required to cause 50% mortality) of 33.1 h compared to B. bassiana (46.7 h), a steeper probit slope (5.4 ± 0.3), and strong model fit (R² = 0.95), indicating rapid and synchronized mortality. Endophytic colonization varied across host species and application methods, with soil incorporation consistently outperforming foliar inoculation. Maximum colonization (82.5%) was recorded in Tecomella undulata and exceeded 80% in Azadirachta indica under M. anisopliae. Biochemical analyses revealed significant increases in protein (up to 3.5 mg g⁻¹), phenols (3.7 mg g⁻¹), and tannins (2.7 mg g⁻¹). Activity of defense enzymes was significantly enhanced, with catalase reaching 263.5 U mL⁻¹, while Phenylalanine ammonia-lyase and Tyrosine ammonia-lyase exceeded 170 and 198 U mL⁻¹, respectively, indicating activation of antioxidant and phenylpropanoid pathways. Molecular docking analysis further revealed strong interactions between fungal metabolites and termite cellulase, with Bassianin (−8.4 kcal mol⁻¹) and Tenellin (−8.1 kcal mol⁻¹) showing the highest binding affinities. These findings highlight the combined biochemical and molecular mechanisms underlying fungal-mediated termite suppression and plant defense induction, and future research should prioritize transcriptomic validation, rhizosphere microbiome interactions, formulation optimization, and long-term multi-location field evaluation to support sustainable termite management strategies. Full article

Pathogen manipulation of host behavior is a widespread evolutionary strategy to enhance its transmission, yet whether different pathogens elicit distinct behavioral and molecular responses in the same host remains poorly understood. We performed parallel behavioral assays and comparative transcriptomic analyses on third-instar Lymantria dispar larvae infected with Lymantria dispar multiple nucleopolyhedrovirus (LdMNPV, virus), Staphylococcus aureus (bacterium) and Metarhizium anisopliae (fungus). Climbing height was recorded over 72 h post-infection, and gene expression pattern was profiled using RNA-seq at 72 h. Only LdMNPV infection induced significant, sustained upward climbing behavior among the three pathogen infection groups. All three pathogens activated Toll and IMD immune pathways, but LdMNPV triggered substantially broader transcriptomic reprogramming. Notably, the virus specifically upregulated multiple energy metabolism pathways (nicotinate/nicotinamide metabolism, pyruvate metabolism, TCA cycle and oxidative phosphorylation) and the neuroactive ligand-receptor interaction pathway—a pattern absent in bacterial and fungal infections. LdMNPV drove tree-top disease through a virus-specific, multi-system manipulation strategy that couples metabolic activation with neural signaling modulation. This comparative study reveals fundamental differences in behavioral manipulation across pathogen kingdoms and provides candidate pathways for functional validation. Full article

(1) Background: Potentially arthropod-pathogenic and plant-associated Metarhizium fungi are of high interest for basic research, biological pest control and plant growth promotion. Unambiguous species delineation enabling the taxonomic assignment of new isolates and the identification of new Metarhizium species is of crucial importance for both research and application. Recently, the new species Metarhizium hybridum and Metarhizium parapingshaense were introduced on the basis of phylogenomic studies. (2) Methods: Neighbor- joining and Bayesian inference-based phylogenetic reconstruction of ribosomal intergenic spacer (rIGS) sequences were used to critically evaluate new species introductions. A species-discriminating diagnostic PCR tool for Metarhizium was adapted to M. hybridum and M. parapingshaense. GenBank database mining was performed to identify cryptic descriptions of the new species. (3) Results: The introduction of M. hybridum and M. parapingshaense was corroborated by rIGS sequence comparison. Data mining revealed cryptic first descriptions of M. hybridum from Canada, China, Colombia, Costa Rica, Cuba, Honduras, Mexico, New Zealand, the USA and the Philippines, and of M. parapingshaense from China, India, Japan, the Philippines and South Korea. (4) Conclusions: Results support the reliability of rIGS as a single taxonomic marker for species-level identification of Metarhizium fungi. Species-discriminating diagnostic PCR was successfully adapted to enable the sequencing-independent identification of the confirmed new species M. hybridum and M. parapingshaense. Full article

The management of Culex pipiens (Diptera: Culicidae), key vectors of arboviruses like West Nile virus, necessitates sustainable alternatives to chemical insecticides. This study screened indigenous entomopathogenic fungi (EPF) from forest soils in Achaia, Greece, for their larvicidal efficacy against Cx. pipiens biotype molestus. Fifteen fungal isolates were obtained via insect baiting and identified as Beauveria and Metarhizium species. A comprehensive bioassay at 1 × 10⁸ conidia mL⁻¹ revealed significant variation in pathogenicity after 72 h. Two isolates, Beauveria bassiana (BB) (Hypocreales: Cordycipitaceae) and Metarhizium anisopliae (K3(1)) (Hypocreales: Clavicipitaceae), exhibited the highest virulence among the tested isolates, each causing 60% mortality with a rapid median lethal time (LT₅₀) of ~18.5 h. Survival analysis, Cox modeling, and non-linear kinetic modeling (Gompertz/Richards) classified three distinct virulence clusters: high/rapid, moderate/consistent, and low/delayed. A pathogenicity network analysis and a composite virulence index further validated BB and K3(1) as the most effective candidates. These results demonstrate the high isolate specificity of fungal efficacy and underscore the importance of screening local fungal diversity. The identified high-virulence isolates represent promising, environmentally sound candidates for the development of targeted biopesticides. Future research should focus on formulation for aquatic environments and integration into resistance-resilient integrated vector management programs. Full article

Empoasca onukii severely damages tea plants as a major sap-sucking pest, leading to the increasing adoption of biopesticides as a sustainable alternative to chemical control. However, existing research has largely focused on the final lethal effects of these agents, while their short-term interference patterns on pest feeding behavior remain unclear. In this study, six biopesticides—azadirachtin, matrine, Beauveria bassiana, Metarhizium anisopliae CQMa421, Mamestra brassicae nucleopolyhedrovirus (MbNPV), and Bacillus thuringiensis (Bt)—were evaluated using the electrical penetration graph (EPG) technique to precisely analyze their interference on the short-term (6 h) feeding behavior of E. onukii, alongside field trials to validate control efficacy. EPG analysis revealed that different types of biopesticides significantly disrupted feeding in distinct ways. The two botanical pesticides and CQMa421 mainly prolonged the non-probing phase (waveform Np) and reduced active non-phloem feeding (C waveform) (p < 0.05); Bt and B. bassiana significantly extended the resting phase (waveform R) and decreased the frequency of passive phloem feeding (waveform E) (p < 0.05), whereas MbNPV exhibited a combined effect, simultaneously prolonging both Np and R waveforms while reducing waveform C (p < 0.05). Field trials showed that all tested treatments achieved complete control (100%) at 21 days post-application. Moreover, across a wide range of concentrations, they all demonstrated excellent and stable control performance. These findings provide diverse agent options for the green control of E. onukii in tea plantations and lay a foundation for constructing a green integrated pest management system centered on biological control for tea plant pests. Full article

The tea geometrid (Ectropis grisescens) is a significant pest in Chinese tea plantations. Although Metarhizium anisopliae serves as an environmentally friendly biocontrol agent against E. grisescens, the molecular mechanisms underlying the insect’s immune response remain unclear. This study investigates changes in immunity-related genes and metabolites in E. grisescens larvae infected with a virulent strain of M. anisopliae through transcriptome sequencing and metabolome analysis. We identified 2409 differentially expressed genes (DEGs) at 48 h post-infection, with 1611 genes up-regulated. GO analysis revealed that 119 DEGs were significantly enriched in immune-related processes. Additionally, 1860 differentially accumulated metabolites (DAMs) were detected, including 652 up-regulated and 1208 down-regulated metabolites, with 236 significantly enriched in 82 KEGG pathways. These findings indicate the activation of immunity-related and detoxifying enzyme-related genes, providing new insights into the physiological and biochemical responses of insects to biopesticides and potential targets for controlling tea geometrid. Full article

Locusta migratoria manilensis (Meyen) is a highly destructive insect pest worldwide. However, excessive reliance on insecticides has resulted in significant environmental pollution. Biocontrol complexes combine two or more BCAs to address the limitations of individual agents. However, biocontrol complex for locust control has been rarely reported. Here, we propose BioControl 3.0, which integrates Metarhizium anisopliae var. acridum (Driver and Milner) and Carabus smaragdinus (Fischer von Waldheim) for locust control. We evaluated this system through a series of laboratory bioassays and semi-field cage experiments, comparing single-agent applications, sequential combinations (BioControl 2.0), and predator-mediated delivery (BioControl 3.0), and quantified locust mortality and interaction effects between predation and infection We found that M. anisopliae caused >85% mortality of locust nymphs at 1 × 10⁸ conidia/mL (LT₅₀ ≈ 6 days) while exhibiting negligible virulence toward C. smaragdinus. BioControl 2.0 (sequential application) increased mortality compared to single agents. However, this approach revealed a significant negative interaction between predation and infection, which limited the total control efficacy. BioControl 3.0 (predator-vectored fungus) achieved the highest corrected mortality, with predation and infection acting independently and additively (no detectable antagonistic interaction). By leveraging a predatory vector, BioControl 3.0 decouples negative interaction and harnesses dual biotic pressures, offering a cost-effective, environmentally benign alternative to conventional locust control. Our findings provide a blueprint for designing integrated predator-pathogen complexes and optimizing deployment strategies for sustainable management of locust outbreaks. Full article

The two-spotted spider mite, Tetranychus urticae Koch, is a major agricultural pest whose control is increasingly constrained by resistance to synthetic acaricides. This study evaluated the long-term field efficacy of three commercial entomopathogenic fungal (EPF) biopesticides—Velifer^® (Beauveria bassiana), Metab^® (B. bassiana + Metarhizium anisopliae), and Botanigard^® (B. bassiana)—against larval and protonymph stages of T. urticae on two host plants, Italian cypress (Cupressus sempervirens) and sweet orange (Citrus sinensis). Two foliar applications were conducted during the 2023 growing season (25 May and 25 July), and mite populations were monitored for 140 days after the final application. A randomized complete block design was used, and efficacy was calculated using the Henderson–Tilton formula. All EPF treatments significantly reduced mite populations compared with the untreated control throughout the monitoring period. Velifer consistently achieved the highest suppression of larval populations, particularly on C. sinensis, with efficacy comparable to the chemical standard. Botanigard showed more gradual but sustained population reduction over time, whereas Metab exhibited lower but stable efficacy in all cases. Treatment performance was strongly influenced by host plant species and mite developmental stage, with larvae consistently more susceptible than protonymphs. On C. sinensis, Velifer achieved the highest larval suppression (84.6%), comparable to the chemical standard abamectin, while Botanigard and Velifer were most effective on C. sempervirens. Survival analysis confirmed isolate- and host-dependent differences in hazard effects over time. These results demonstrate that EPF-based products can provide sustained, long-term suppression of T. urticae under field conditions, supporting their integration into integrated pest management programs. Full article

The tomato leafminer, Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae), is one of the most destructive pests of tomato crops worldwide. Its high reproductive potential and increasing resistance to conventional insecticides have made the development of sustainable management strategies essential. Biological control using entomopathogenic fungi (EPF), particularly when established as endophytes, has emerged as a promising approach. This study investigated the endophytic colonization capacity and greenhouse performance of three commercially available EPF formulations: Beauveria bassiana (Velifer^®), Lecanicillium lecanii (Lecan^®), and a Beauveria bassiana–Metarhizium anisopliae mixture (Metab^®), for the suppression of T. absoluta in tomato. Our experiment was conducted under commercial greenhouse conditions using soil drench applications at manufacturer-recommended doses. Endophytic colonization was assessed through surface-sterilized leaf assays, while pest suppression was evaluated via weekly measurements of larval mine length, infestation incidence, and survival dynamics. B. bassiana (Velifer^®) exhibited the highest endophytic colonization frequency and consistently reduced mine length and infestation levels compared with untreated plants. Survival analysis using Cox proportional hazards revealed significant reductions in infestation risk for Velifer^® (hazard ratio, HR = 0.420), Metab^® (HR = 0.480), and Lecan^® (HR = 0.599), relative to the negative control, whereas the chemical positive control provided the strongest overall suppression (HR = 0.287). Our findings demonstrate that commercial EPF formulations can significantly reduce T. absoluta infestation under greenhouse conditions and represent a valuable component of integrated pest management programs. Full article

The search for sustainable pest management alternatives has intensified due to the risks of chemical pesticides. Entomopathogenic fungi and plant extracts, rich in insecticidal secondary metabolites, are among the most promising approaches. Integrating these agents can enhance complementary mechanisms and reduce environmental impact. This study evaluated the insecticidal potential of fungi produced by submerged fermentation (Beauveria bassiana, Metarhizium anisopliae, Trichoderma asperelloides, Isaria javanica, and Cordyceps fumosorosea) applied alone and combined with Trichilia claussenii extract against Euschistus heros and Spodoptera frugiperda. Fermentation showed good fungal adaptation and high sporulation, especially B. bassiana (8.33 × 10⁸ spores mL⁻¹) and T. asperelloides (9.42 × 10⁷ spores mL⁻¹). Adding the plant extract increased colony-forming units, notably for M. anisopliae (7.40 × 10⁷ CFU mL⁻¹) and B. bassiana (1.55 × 10⁸ CFU mL⁻¹). In bioassays, cell suspensions were more effective than isolated metabolites, reaching 97.8% mortality for E. heros and 91.5% for S. frugiperda with B. bassiana plus extract. These results indicate that combining entomopathogenic fungi with T. claussenii extract is a promising strategy for developing efficient and sustainable biopesticides, contributing directly to integrated pest management practices with reduced environmental impact. Full article

Entomopathogenic fungi are promising alternatives to synthetic insecticides for the control of vector species, notably the arbovirus vector, Aedes aegypti. The influence of intrinsic mosquito midgut microbiota on host susceptibility to fungal infection and subsequent physiological processes remains poorly understood. Here we treated female Ae. aegypti with the broad-spectrum antibiotic carbenicillin to reduce gut bacterial populations, then exposed them to Metarhizium anisopliae conidia. Female Ae. aegypti offered carbenicillin and then sprayed with fungi had significantly lower survival rates (38.9% ± 1.15) compared to non-antibiotic-treated mosquitoes sprayed with fungus (68.9% ± 0.58). To monitor the kinetics of microbial community recovery, mosquitoes were challenged with conidia at 0, 3, 6, and 9 days following antibiotic removal from the diet. Reduced survival persisted through the 6-day period (survival rates 37.8% to 45.6%), with a significant increase in survival observed 9 days post-antibiotic removal (58.9% vs. control 63.3%), which coincided with recovery of gut bacterial populations. Additionally, antibiotic and fungal treatments reduced egg production, larval eclosion, and pupal formation. These results demonstrate that gut bacteria contribute to mosquito defense against fungal pathogens and support normal reproductive and developmental functions. Understanding the interplay between gut microbiota and entomopathogenic fungi may enhance biological control approaches. Full article

Chemical defenses help insects resist pathogen infection. The volatile compositions, emission patterns, and external immune functions of the red palm weevil (RPW), a major invasive pest now established in numerous countries including the Mediterranean Basin, North Africa, Middle East, and parts of Latin America and the Caribbean, are largely unknown. In this study, we examined RPW larval volatiles, analyzing their emission patterns across developmental stages and under pathogen stress from feeding Metarhizium anisopliae. RPW larvae shared a number of volatile components across stages, but the emission dynamics were significantly different. These volatile chemicals were primarily alcohols, phenols and aromatic hydrocarbons, and styrene was the predominant volatile. Furthermore, pathogen stress induced distinct volatile profiles: phenylacetaldehyde unidirectionally decreased, whereas n-nonanol, 4-ethylguaiacol, 2-phenylethanol, hexanal, and benzophenone levels increased. Antimicrobial analysis showed that these upregulated compounds broadly inhibited fungal and bacterial growth. Therefore, our results illustrated the immune defense role of RPW larval volatiles and their potential bioactive compounds, including n-nonanol, 4-ethylguaiacol, 2-phenylethanol, hexanal, and benzophenone. Full article