UNASSIGNED: As a genus within the Clavicipitaceae, Metarhizium exhibits rich morphological and ecological diversity, with a wide distribution and a variety of hosts. Currently, sixty-eight species of Metarhizium have been described.
UNASSIGNED: A new species of Metarhizium, M.puerense (Hong Yu bis), was described in Pu\'er City, Yunnan Province, south-western China. Based on morphological characteristics and multilocus phylogenetic analyses, Metarhiziumpuerense was confirmed to be phylogenetically related to M.album, but was clearly separated and formed a distinct branch. In contrast, the host of Metarhiziumalbum was plants and leafhoppers and that lepidopteran larvae were the host of M.puerense. The diagnostic features of M.puerense were solitary to multiple stromata and smooth-walled, cylindrical with rounded apices conidia.

UNASSIGNED: The tomato, Solanum lycopersicum L., is one of the most important horticultural crops that can be consumed fresh or after being processed worldwide. The tomato leaf miner (Tuta absoluta) is one of the most devastating pest to tomato plants due to its mine-feeding nature in the mesophyll tissue of the plant. Fungal entomopathogens can exist naturally in plants as an asymptote. This study aimed to detect the endophytic colonization of Beauveria bassiana and Metarhizium robertsii within tomato plants via artificial inoculation and their virulence effects on Tuta absoluta.
UNASSIGNED: Isolates with the highest percent germination and virulence against T. absoluta were selected for endophytic evaluation within tomato plants by different artificial inoculation techniques.
UNASSIGNED: This study revealed that, isolates with the highest percent germination and virulent to Tuta absoluta had the potential to colonize tomato plants. The result showed that, the maximum mortality rate (97.5%) of Tuta absoluta larvae was achieved by Metarhizium robertsii isolate K-61 at a concentration of 1x10 8conidial/ml at 7 days post inoculated. However, the highest cumulative mortality (100%) was recorded by Beauveria bassiana isolate APPRC-27 at 10 days post inoculated through the direct contact method. The highest endophytic colonization was registered by isolate APPRC-27 (76.67%) at 7 days post-inoculated using the leaf spray technique, but it declined to 11.67% after 28 days of inoculated. In the case of the seedling inoculation technique, the highest endophytic colonization was obtained in the root tissues of tomatoes at 28 days of inoculated by isolate K-61.
UNASSIGNED: This study revealed that the leaf spray inoculation technique was the most effective method, followed by seedling inoculation, for the deployment of Beauveria bassiana and Metarhizium robertsii endophytes in tomato plant tissues. Therefore, virulent Beauveria bassiana and Metarhizium robertsii, are promising bioagents for the control of Tuta absoluta if deployed as endophytes.

OBJECTIVE: The Colorado potato beetle, Leptinotarsa decemlineata (Say) (Coleoptera: Chrysomelidae) is the most widespread insect pest that causes major economic losses, especially on potatoes. Due to heavy insecticide use, this species now resists most pesticides, posing a significant control challenge. Frequent pesticide application also harms non-target organisms, the environment, and human health. Hence, utilizing biocontrol agents like entomopathogenic fungi (EPF) offers a viable alternative for pest management. The aim of this study was to identify and characterize new EPF strains isolated from soil samples and evaluate their efficacy against adult L. decemlineata under laboratory conditions.
RESULTS: Soil samples were collected in potato fields or uncultivated areas adjacent to the field in the Czech Republic and the EPF strains were isolated using a modified Tenebrio bait method. A total of 20 fungal strains were isolated and identified using morphological and molecular markers based on the 28S rRNA, ITS, and elongation factor 1-alpha gene sequences as Beauveria bassiana (Bals.-Criv.) Vuill., Beauveria brongniartii (Sacc.) Petch, and Cordyceps fumosorosea (Wize) Kepler, B. Shrestha & Spatafora (Hypocreales: Cordycipitaceae), Purpureocillium lilacinum (Thom.) Luangsa-ard, Houbraken, Hywel-Jones & Samson (Hypocreales: Ophiocordycipitaceae), Metarhizium brunneum (Petch), and Metarhizium robertsii Bisch., Rehner & Humber (Hypocreales: Clavicipitaceae). The bioassays revealed high variability among virulence of these strains against L. decemlineata with the shortest median time to death (LT50 = 5 days) in M. robertsii strain MAN3b.
CONCLUSIONS: Results shown that some EPF strains, particularly those of genera Metarhizium, can be promising biocontrol agents against the Colorado potato beetle.

BACKGROUND: Lyme disease continues to expand in Canada and the USA and no single intervention is likely to curb the epidemic.
METHODS: We propose a platform to quantitatively assess the effectiveness of a subset of Ixodes scapularis tick management approaches. The platform allows us to assess the impact of different control treatments, conducted either individually (single interventions) or in combination (combined efforts), with varying timings and durations. Interventions include three low environmental toxicity measures in differing combinations, namely reductions in white-tailed deer (Odocoileus virginianus) populations, broadcast area-application of the entomopathogenic fungus Metarhizium anisopliae, and fipronil-based rodent-targeted bait boxes. To assess the impact of these control efforts, we calibrated a process-based mathematical model to data collected from residential properties in the town of Redding, southwestern Connecticut, where an integrated tick management program to reduce I.xodes scapularis nymphs was conducted from 2013 through 2016. We estimated parameters mechanistically for each of the three treatments, simulated multiple combinations and timings of interventions, and computed the resulting percent reduction of the nymphal peak and of the area under the phenology curve.
RESULTS: Simulation outputs suggest that the three-treatment combination and the bait boxes-deer reduction combination had the overall highest impacts on suppressing I. scapularis nymphs. All (single or combined) interventions were more efficacious when implemented for a higher number of years. When implemented for at least 4 years, most interventions (except the single application of the entomopathogenic fungus) were predicted to strongly reduce the nymphal peak compared with the no intervention scenario. Finally, we determined the optimal period to apply the entomopathogenic fungus in residential yards, depending on the number of applications.
CONCLUSIONS: Computer simulation is a powerful tool to identify the optimal deployment of individual and combined tick management approaches, which can synergistically contribute to short-to-long-term, costeffective, and sustainable control of tick-borne diseases in integrated tick management (ITM) interventions.

Similar to the physiological importance of gut microbiomes, recent works have shown that insect ectomicrobiotas can mediate defensive colonization resistance against fungal parasites that infect via cuticle penetration. Here we show that engineering the entomopathogenic fungus Metarhizium robertsii with a potent antibacterial moricin gene from silkworms substantially enhances the ability of the fungus to kill mosquitos, locusts, and two Drosophila species. Further use of Drosophila melanogaster as an infection model, quantitative microbiome analysis reveals that engineered strains designed to suppress insect cuticular bacteria additionally disrupt gut microbiomes. An overgrowth of harmful bacteria such as the opportunistic pathogens of Providencia species is detected that can accelerate insect death. In support, quantitative analysis of antimicrobial genes in fly fat bodies and guts indicates that topical fungal infections result in the compromise of intestinal immune responses. In addition to providing an innovative strategy for improving the potency of mycoinsecticides, our data solidify the importance of both the ecto- and endo-microbiomes in maintaining insect wellbeing.

Symbiotic and pathogenic microorganisms such as bacteria and fungi represent promising alternatives to chemical insecticides to respond to the rapid increase of insecticide resistance and vector-borne disease outbreaks. This study investigated the interaction of two strains of Wolbachia, wAlbB and wAu, with the natural entomopathogenic fungi from Burkina Faso Metarhizium pingshaense, known to be lethal against Anopheles mosquitoes. In addition to showing the potential of Metarhizium against African Aedes aegypti wild-type populations, our study shows that the wAlbB and wAu provide a protective advantage against entomopathogenic fungal infections. Compared to controls, fungal-infected wAu and wAlbB-carrying mosquitoes showed higher longevity, without any significant impact on fecundity and fertility phenotypes. This study provides new insights into the complex multipartite interaction among the mosquito host, the Wolbachia endosymbiont and the entomopathogenic fungus that might be employed to control mosquito populations. Future research should investigate the fitness costs of Wolbachia, as well as its spread and prevalence within mosquito populations. Additionally, evaluating the impact of Wolbachia on interventions involving Metarhizium pingshaense through laboratory and semi-field population studies will provide valuable insights into the effectiveness of this combined approach.

Metarhizium anisopliae is an effective biopesticide for controlling Aphis citricola, which has developed resistance to many chemical pesticides. However, the powerful immune system of A. citricola has limited the insecticidal efficacy of M. anisopliae. The co-evolution between insects and entomogenous fungi has led to emergence of new antifungal immune genes, which remain incompletely understood. In this study, an important immune gene Sgabd-2 was identified from A. citricola through transcriptome analysis. Sgabd-2 gene showed high expression in the 4th instar nymph and adult stages, and was mainly distributed in the abdominal region of A. citricola. The recombinant protein (rSgabd-2) exhibited no antifungal activity but demonstrated clear agglutination activity towards the conidia of M. anisopliae. RNA interference of Sgabd-2 by dsRNA feeding resulted in decreased phenoloxidase (PO) activity and weakened defense for A. citricola against M. anisopliae. Simultaneous silence of GNBP-1 and Sgabd-2 effectively reduced the immunity of A. citricola against M. anisopliae more than the individual RNAi of GNBP-1 or Sgabd-2. Furthermore, a genetically engineered M. anisopliae expressing double-stranded RNA (dsSgabd-2) targeting Sgabd-2 in A. citricola successfully suppressed the expression of Sgabd-2 and demonstrated increased virulence against A. citricola. Our findings elucidated Sgabd-2 as a critical new antifungal immune gene and proposed a genetic engineering strategy to enhance the insecticidal virulence of entomogenous fungi through RNAi-mediated inhibition of pest immune genes.

The tropical climate in Malaysia provides an ideal environment for the rapid proliferation of Aedes mosquitoes, notably Aedes aegypti and Aedes albopictus, prominent vectors of dengue fever. Alarmingly, these species are increasingly developing resistance to conventional pesticides. This study aimed to evaluate the efficacy of Metarhizium anisopliae isolate HSAH5 spores, specifically on conidia (CO) and blastospores (BL), against Ae. albopictus larvae. The study centered on evaluating their pathogenic effects and the resultant changes in protein expression. Spore suspensions with varying concentrations were prepared for larvicidal bioassays, and protein expressions were analysed using liquid chromatography-mass spectrometry. Subsequently, protein annotation and network analysis were conducted to elucidate infection mechanisms and the proteomic response. Based on the lethal concentrations and time frames, CO exhibited faster larval mortality than BL at lower concentrations. Despite this, both spore types demonstrated comparable overall pathogenic effects. Results from the proteomic profiling revealed 150 proteins with varied expressions following exposure to Ae. albopictus extract, shedding light on distinct infection strategies between the spores. Gene Ontology enrichment and network analysis illustrated the diverse metabolic adaptations of M. anisopliae and interactions with mosquito larvae. This highlighted the complexity of host-pathogen dynamics and the significance of biosynthetic processes, energy storage, and cellular interaction pathways in disease progression. The BL network, consisting 80 proteins and 74 connections, demonstrates the intricate fungal mechanisms triggered by host stimuli. Conversely, the CO network, though smaller, displayed notable interconnectivity and concentrated involvement at the cell periphery, suggesting a deliberate strategy for initial host contact. This study offers valuable insights into proteome dynamics of M. anisopliae\'s BL and CO for managing mosquito populations and combating disease transmission, thereby significantly advancing public health and environmental conservation efforts.

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.

Entomopathogenic fungi capable of establishing mutualistic endophytic relationships with plants have a tremendous potential as biocontrol agents of insect pests. While fungi have long played an important and highly effective role in pest suppression, the utility of endophytic entomopathogenic fungi in pest management is a relatively new and emerging topic of biocontrol. Here we discuss the relevance of endophytic fungi to plant health in general, synthesize the current knowledge of the effectiveness of endophytic entomopathogenic fungi against diverse insect pests, discuss the indirect plant-mediated effects of endophytic entomopathogenic fungi on arthropods, and describe the diverse benefits of endophytic fungi to plants that are likely to affect herbivores and plant pathogens as well. Lastly, we consider major challenges to incorporating endophytic entomopathogenic fungi in biocontrol, such as their non-target effects and field efficacy, which can be variable and influenced by environmental factors. Continued research on endophyte-insect-plant-environment interactions is critical to advancing our knowledge of these fungi as a sustainable pest management tactic. © 2024 The Author(s). Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.