Search Results (20)

Pyrethroids are chemical insecticides used on a large scale in agriculture, horticulture, and forest protection. In order to reduce their use in IPM, alternative methods of controlling insect pests are introduced, such as the use of biopesticides based on entomopathogenic fungi (EPF). Species of the Beauveria genus are characterized by a very broad spectrum of action, which is why they are often used to produce preparations based on EPF. The aim of the study was to determine the effect of different doses of tested pyrethroids on the colony growth and metabolic activity of EPF from the Beauveria genus. In vitro, the effect of three pyrethroids (deltamethrin, λ-cyhalothrin, and α-cypermethrin) added to SDA medium at a dose 10 times lower than the recommended field dose (A), the recommended field dose (B), and 10 times higher than the recommended field dose (C) on colony growth and metabolic activity of B. bassiana and B. brongniartii was tested. The research carried out showed that pyrethroid insecticides used in the experiment showed various toxic effects towards the tested EPF of the genus Beauveria. The studies conducted showed that on the 20th day of the observation, λ-cyhalothrin used in the recommended field dose limited the growth of B. bassiana to the least extent in relation to the other tested pyrethroids. However, with respect to the fungus B. brongniartii, no toxic effect of this pyrethroid was found. Based on the results obtained, it was found that λ-cyhalothrin used in the recommended field dose and 10 times lower than recommended significantly increased the metabolic activity of B. bassiana. In relation to the B. brongniartii strain, detlamethrin used in each of the tested concentrations significantly affected its viability. Full article

Spodoptera frugiperda (Lepidoptera: Noctuidae) is a migratory agricultural pest that is devastating on a global scale. Beauveria bassiana is a filamentous entomopathogenic fungus that has a strong pathogenic effect on Lepidoptera pests but little is known about the microbial community in the host gut and the dominant populations in fungus-infected insects. B. bassiana AJS91881 was isolated and identified from the infected larvae of Spodoptera litura. The virulence of AJS91881 to the eggs, larvae, pupae and adults of S. frugiperda was measured. Moreover, the gut microbial community diversity of healthy and fungus-infected insects was analyzed. Our results showed that after treatment with B. bassiana AJS91881, the egg hatching rate, larval survival rate and adult lifespan of the insects were significantly reduced, and the pupae rigor rate was significantly increased compared to that of the control group. Additionally, the gut microbial community was reconstructed after B. bassiana infection. At the phylum and genus level, the relative abundance of the Proteobacteria and Serratia increased significantly in the B. bassiana treatment group. The KEGG function prediction results showed that fungal infection affected insect gut metabolism, environmental information processing, genetic information processing, organism systems and cellular processes. Fungal infection was closely related to the metabolism of various substances in the insect gut. Serratia marcescens was the bacterium with the highest relative abundance after infection by B. bassiana; intestinal bacteria S. marcescens inhibited the infection of insect fungi B. bassiana against the S. frugiperda. The presence of gut bacteria also significantly reduced the virulence of the fungi against the insects when compared to the group with the larvae fed antibiotics that were infected with fungal suspension (Germfree, GF) and healthy larvae that were infected with fungal suspension prepared with an antibiotic solution (+antibiotic). In conclusion, the reconstruction of the insect intestinal bacterial community is an indispensable link for understanding the pathogenicity of B. bassiana against S. frugiperda. Most importantly, in the later stage of fungal infection, the increased abundance of S. marcescens in the insect intestine inhibited the virulence of B. bassiana to some extent. The findings aid in understanding changes in the gut microbiota during the early stages of entomopathogenic fungal infection of insects and the involvement of insect gut microbes in host defense mediated by pathogenic fungal infection. This study is also conducive to understanding the interaction between entomopathogenic fungi, hosts and gut microbes, and provides a new idea for the joint use of entomopathogenic fungi and gut bacteria to control pests. Full article

The species composition of fungal pathogens among three of the most harmful locust species (Asian migratory locust, Moroccan locust and Italian locust) in the southern Russian Federation was studied. Insects were sampled in 20 locations of seven federal subjects of the Russian Federation (Republic of Dagestan, Republic of Kalmykia; Krasnodar and Stavropol Krai; Astrakhan, Volgograd and Rostov Regions). Forty-five isolates belonging to the fungal genus Beauveria were collected, particularly isolates of B. bassiana and B. pseudobassiana. B. bassiana was the most prevalent, constituting 98% of the samples, and could be differentiated into three clades, as evidenced by the TEF gene and intergenic spacer Bloc. Clade 1, represented by the reference isolate ARSEF 2040, was most abundant (61%), and Clade 2, represented by ARSEF 1811, had lower abundance (27%). The remaining isolates either belonged to the genetically distinct Clade 3, represented by ARSEF 1564, or were found to fall outside the major lineages. The frequency of infection in locust populations was variable and tended to increase under conditions unfavorable for the insects. The vast majority of Beauveria isolates from locusts were highly virulent in this insect group. Full article

The genus Beauveria includes important entomopathogenic and endophytic fungi; among them, Beauveria bassiana is the most studied species. However, there is little knowledge regarding their antimicrobial activity. The current research has been conducted to evaluate the in vitro antagonistic activity of B. bassiana and the antimicrobial efficacy of its Exo and Endo metabolites against Bacillus cereus, B. megaterium, Clavibacter michiganensis (Gram positive bacteria, G+ve), Xanthomonas campestris, Pseudomonas aeruginosa and P. fluorescence (Gram negative bacteria, G−ve). In addition, solid-phase microextraction (SPME) was coupled with Gas Chromatography-Mass Spectrometry (GC/MS) to qualitatively measure the volatile organic compounds’ (VOCs) metabolic profile of the most efficient studied isolate of B. bassiana. The obtained results showed that the isolate UniB2439-3 has a promising antibacterial effect against most of the studied target bacteria. An SPME-GC/MS analysis of VOCs revealed the presence of ethanol, butanal,2-methyl, 2,4-dimethyl-1-heptene, octane, 4-methyl and β-elemene as the dominant bioactive compounds. The results demonstrated that the efficient isolate of B. bassiana can be potentially used as a biocontrol agent against several bacteria, especially G+ve ones. Full article

The genus Amphichorda has been recently re-erected as an independent lineage from Beauveria, circumscribed within Cordycipitaceae. However, its phylogenetic relationships with other members of this family remain obscure. In our ongoing efforts to expand the knowledge on the diversity of culturable ascomycetes from the Mediterranean Sea, we isolated several specimens of Amphichorda. Preliminary sequence analyses revealed great phylogenetic distance with accepted Amphichorda species and a close relation to Onychophora coprophila. Onychophora is a monotypic genus of enteroblastic conidiogenous cells, presumably related to Acremonium (Bionectriaceae), while Amphichorda exhibits holoblastic conidiogenesis. Here, we examine representative strains of Amphichorda species to elucidate the taxonomy of the genus and the aforementioned fungi, combining morphological, ultrastructure and multi-locus phylogenetic analyses (ITS, LSU, tef1, BenA). The results revealed Amphichorda as a member of the Bionectriaceae, where its asexual morphs represent a transition between enteroblastic and holoblastic conidiogenesis for this group of fungi. We also describe and illustrate Amphichorda littoralis sp. nov. and propose Amphichorda coprophila comb. nov. In addition, we establish key phenotypic features to distinguish Amphichorda species and demonstrate the higher salt tolerance degree of A. littoralis, consistent with its marine origin. This work provides a comprehensive framework for future studies in the genus. Full article

Oaxaca, Mexico, is home to over 30 species of the genus Agave, and its cultivation is of great economic and social importance for the mezcal industry, which depends on its production. The incidence of the pest Scyphophorus acupunctatus causes severe losses and damage. Agrochemicals are used for its control, but a viable alternative is microbial control. The objectives of this study were to determine the natural occurrence of the entomopathogenic fungi (EPF) Beauveria bassiana and Metarhizium anisopliae, isolated from S. acupunctatus in agave crops, and to evaluate the effect of vegetable oil in water emulsions containing conidia from the native fungi against adults of S. acupunctatus under laboratory conditions. Viability of the fungal isolates was determined at a concentration of ×10⁸ conidia/mL in avocado (Persea americana), mamey (Mammea americana), chia (Salvia hispanica), or olive (Olea europaea) oil in water emulsions at two concentrations, 20% and 40%. The most effective oil emulsion on S. acupunctatus adults was determined under laboratory conditions. Naturally occurring fungi in 900 field collected insects was 2.21%. Ninety-six hours after preparing a 20% emulsion in P. americana oil, B. bassiana and M. anisopliae had a viability of 75% and 66.5%, respectively, while the control conidia suspended in distilled water remained viable for only 48 h. Twenty-four hours after applying M. anisopliae conidia in a 40% P. americana oil emulsion, effectiveness was 100% on S. acupunctatus, followed by M. anisopliae in 20% P. americana oil emulsion with 75% effectiveness. At 72 h post-application, all fungus in oil emulsions achieved an accumulated insect mortality of 100%, while the control showed no effect on adult S. acupunctatus. The most promising combination was 40% P. americana oil emulsion, which achieved 50% viable B. bassiana or M. anisopliae conidia up to 96 h after preparation, and its accumulated effectiveness on S. acupunctatus adults was 87.5% after 24 h. Full article

The Dioryctria genus contains several destructive borer pests that are found in coniferous forests in the Northern Hemisphere. Beauveria bassiana spore powder was tested as a new method of pest control. In this study, Dioryctria sylvestrella (Lepidoptera: Pyralidae) was used as the object. A transcriptome analysis was performed on a freshly caught group, a fasting treatment control group, and a treatment group inoculated with a wild B. bassiana strain, SBM-03. Under the conditions of 72-h fasting and a low temperature of 16 ± 1 °C, (i) in the control group, 13,135 of 16,969 genes were downregulated. However, in the treatment group, 14,558 of 16,665 genes were upregulated. (ii) In the control group, the expression of most genes in the upstream and midstream of the Toll and IMD pathways was downregulated, but 13 of the 21 antimicrobial peptides were still upregulated. In the treatment group, the gene expression of almost all antimicrobial peptides was increased. Several AMPs, including cecropin, gloverin, and gallerimycin, may have a specific inhibitory effect on B. bassiana. (iii) In the treatment group, one gene in the glutathione S-transferase system and four genes in the cytochrome P450 enzyme family were upregulated, with a sharp rise in those that were upregulated significantly. In addition, most genes of the peroxidase and catalase families, but none of the superoxide dismutase family were upregulated significantly. Through innovative fasting and lower temperature control, we have a certain understanding of the specific defense mechanism by which D. sylvestrella larvae may resist B. bassiana in the pre-wintering period. This study paves the way for improving the toxicity of B. bassiana to Dioryctria spp. Full article

Diamondback moth (DBM) is an important horticultural pest worldwide as the larvae of these moths feed on the leaves of cruciferous vegetables. As DBM has developed resistance to more than 100 classes of synthetic insecticides, new biological control options are urgently required. Beauveria species are entomopathogenic fungi recognized as the most important fungal genus for controlling a wide range of agricultural, forestry, and veterinary arthropod pests. Previous research, aimed at developing new Beauveria-based biopesticides for DBM, has focused on screening single isolates of Beauveria bassiana. However, these fungal isolates have individual requirements, which may limit their effectiveness in some environments. This current study separately assessed 14 Beauveria isolates, from a range of habitats and aligned to four different species (Beauveria bassiana, B. caledonica, B. malawiensis, and B. pseudobassiana), to determine the most effective isolate for the control of DBM. Further assays then assessed whether selected combinations of these fungal isolates could increase the overall efficacy against DBM. Six Beauveria isolates (three B. bassiana and three B. pseudobassiana) achieved high DBM mortality at a low application rate with the first documented report of B. pseudobassiana able to kill 100% of DBM larvae. Further research determined that applications of low-virulent Beauveria isolates improved the control of DBM compared to mixtures containing high-virulent isolates. This novel approach increased the DBM pest mortality and shortened the time to kill. Full article

The entomopathogenic fungus Beauveria pseudobassiana strain RGM 2184 can reach a maximum efficacy of 80% against the quarantine pest Lobesia botrana in field assays. In this study, the RGM 2184 genome was sequenced, and genome mining analyses were performed to predict the factors involved in its insecticidal activity. Additionally, the metabolic profiling of the RMG 2184 culture’s supernatants was analyzed by mass spectrometry, and the insecticidal activity from one of these extracts was evaluated in Galleria mellonella larvae. The genome analysis resulted in 114 genes encoding for extracellular enzymes, four biosynthetic gene clusters reported as producers of insecticidal and bactericidal factors (oosporein, beauvericin, desmethylbassianin, and beauveriolide), 20 toxins, and at least 40 undescribed potential biocontrol factors (polyketides and nonribosomal peptides). Comparative genomic analysis revealed that 65–95% of these genes are Beauveria genus-specific. Metabolic profiling of supernatant extracts from RGM 2184 cultures exhibited secondary metabolites such as beauveriolide, oosporein, inflatin C, and bassiatin. However, a number of detected metabolites still remain undescribed. The metabolite extract caused 79% mortality of Galleria mellonella larvae at 28 days. The results of this research lay the groundwork for the study of new insecticidal molecules. Full article

Entomopathogenic fungi (EPF) in Croatian forests are known only from observations of insect cadavers that show obvious signs of disease. To date, their presence in soils has not been investigated. The aim of this study was to investigate their occurrence, diversity, and distribution, and to assess their density in tested soils. Soil samples were collected during 2018, 2019, and 2020 at different localities throughout the country, and analyzed by using a method of isolation of fungi on selective culture media. To assess the density of EPF in tested soils, colonies of individual fungal species were counted and recorded; the results were expressed as the number of colony-forming units (CFU) per gram of dry soil. After morphological and molecular analysis, five entomopathogenic fungal genera were identified: Beauveria spp., Metarhizium spp., Purpureocillium spp., Lecanicillium spp., and Paecilomyces spp. Results also showed that the range of a total EPF colony density in the soil varies from 4 × 10³ to 27.4 × 10³ CFU g⁻¹. The most common were EPF of the genus Beauveria, which were recorded at four of five locations, and at 16 of 25 sampling points, but the highest average number (density) of colonies belonged to the genus Metarhizium. Since this type of research was never conducted in Croatia previously, this is the first evidence that insect pathogenic fungi are present in soils of different natural forest habitats. Such research can be useful in selecting and utilizing entomopathogens that are suitable for biological pest control in certain target areas. Full article

This paper investigates 71 isolates of two genera of entomopathogens, Metarhizium and Beauveria, and a biostimulative genus Trichoderma, for their ability to infect yellow mealworms (Tenebrio molitor) and to stimulate maize (Zea mays) growth. Fungal origin, host, and isolation methods were taken into account in virulence analysis as well. Isolates Metarhizium brunneum (1154) and Beauveria bassiana (2121) showed the highest mortality (100%) against T. molitor. High virulence seems to be associated with fungi isolated from wild adult mycosed insects, meadow habitats, and Lepidopteran hosts, but due to uneven sample distribution, we cannot draw firm conclusions. Trichoderma atroviride (2882) and Trichoderma gamsii (2883) increased shoot length, three Metarhizium robertsii isolates (2691, 2693, and 2688) increased root length and two M. robertsii isolates (2146 and 2794) increased plant dry weight. Considering both criteria, the isolate M. robertsii (2693) was the best as it caused the death of 73% T. molitor larvae and also significantly increased maize root length by 24.4%. The results warrant further studies with this isolate in a tri-trophic system. Full article

The cedar forests of Lebanon have been threatened by the outbreak caused by climate change of a web-spinning sawfly, Cephalcia tannourinensis (Hymenoptera: Pamphiliidae), which negatively impacted the survival of one of the oldest tree species on earth. In this study, we investigated the occurrence of naturally soil-inhabiting entomopathogenic fungi for their role in containing the massive outbreak of this insect. We used a combination of fungal bioexploration methods, including insect bait and selective media. Morphological features and multilocus phylogeny—based on Sanger sequencing of the transcripts encoding the translation elongation factor 1-alpha (TEF-α), RNA polymerase II second largest subunit (RBP2), and the nuclear intergenic region (Bloc) were used for species identification. The occurrence rate of entomopathogenic fungi (EPF) varied with location, soil structure, forest structure, and isolation method. From 15 soil samples positive for fungal occurrence, a total of 249 isolates was obtained from all locations using different isolation methods. The phylogenetic analysis confirmed the existence of two novel indigenous species: Beauveria tannourinensis sp. nov. and Beauveria ehdenensis sp. nov. In conclusion, the present survey was successful (1) in optimizing the isolation methods for EPF, (2) investigating the natural occurrence of Beauveria spp. in outbreak areas of C. tannourinensis, and (3) in characterizing the presence of new Beauveria species in Lebanese cedar forest soil. Full article

Beauveria and Metarhizium fungi are facultative plant endophytes that provide plant growth-stimulating, immunomodulatory, and other beneficial effects. However, little is known about the level of plant colonization by these fungi under natural conditions. We assessed the endophytic colonization of potatoes (Solanum tuberosum) with entomopathogenic fungi at their natural load in soils (10²–10⁴ colony-forming units per g). Microbiological analyses of soils and plant organs, as well as a metagenomic analysis of potato roots and leaves, were conducted in three locations in Western Siberia, consisting of conventional agrosystems and kitchen gardens. The fungi were isolated at a relatively high frequency from unsterilized roots (up to 53% of Metarhizium-positive plants). However, the fungi were sparsely isolated from the internal tissues of roots, stems, and leaves (3%). Among the genus Metarhizium, two species, M. robertsii and M. brunneum, were detected in plants as well as in soils, and the first species was predominant. A metagenomic analysis of internal potato tissues showed a low relative abundance of Beauveria and Metarhizium (<0.3%), and the communities were represented primarily by phytopathogens. We suggest that colonization of the internal tissues of potatoes occurs sporadically under a natural load of entomopathogenic fungi in soils. The lack of stable colonization of potato plants with Beauveria and Metarhizium may be due to competition with phytopathogens. Full article

The natural occurrence of entomopathogenic fungal endophytes in sugarcane (Saccharum officinarum) and in soil samples from sugarcane fields was evaluated in Chikwawa District, southern Malawi. Fungi from soil were isolated by baiting using Galleria mellonella larva. Fungal endophytes were isolated from surface-sterilized plant tissue sections. Forty-seven isolates resembled the genus Beauveria, 9 isolates were Metarhizium, and 20 isolates were Isaria. There was no significant difference in the number and type of fungal isolates collected from soil and from plant tissue. There was, however, a significant difference in the part of the plant where fungal species were isolated, which fungal species were isolated, and the number of fungal species isolated at each location. Phylogenetic analysis of 47 Beauveria isolates based on DNA sequencing of the Bloc intergenic region indicated that these isolates all belonged to B. bassiana and aligned with sequences of B. bassiana isolates of African and Neotropical origin. The Malawian B. bassiana isolates formed a distinct clade. No larvae died from infestation by multiple fungi. To the best of our knowledge, this is the first report of B. bassiana and Isaria spp. occurring naturally as endophytes in sugarcane. Further, it is the first report of B. bassiana, Isaria spp., and Metarhizium spp. in the soil of sugarcane fields in Africa. Full article

The ability of ants to detect and remove conidia-contaminated aphids, aimed at limiting contacts with potentially dangerous entities, is an effective antifungal mechanism to prevent the spread of infection among both their nestmates and aphids, their main suppliers of carbohydrates. However, the spread and the scale of this quarantining behaviour among ants are still scarcely studied. Among seven ant species studied, active usage of quarantining behaviour was found only in Formica ants. The behaviour of Lasius fuliginosus (Latreille) aphid milkers towards Chaitophorus populeti (Panzer) aphids covered with conidia of Beauveria bassiana (Balsamo-Crivelli) Vuillemin was studied in the field. Most aggressive milkers quickly detected and removed conidia-contaminated aphids from the plant, carrying them down and placing them some distance away from the experimental aspen trees. In general, active usage of quarantining behaviour towards conidia-contaminated aphids was found to be not limited to the genus Formica, but typical of L. fuliginosus as well. The response of milkers of L. fuliginosus and Formica s. str. ants to living aphids covered with conidia is quite similar. Removal of most fungus-contaminated aphids from the plant enables these ants to reduce the risk of infection transmission among both their nestmates and aphids. Full article