BACKGROUND: Beauveria bassiana is one of the commercially available entomopathogenic fungi (EPF), and a number of isolates with high virulence and broad host spectrum have been used to control agricultural and forest pests. Although the functional importance of genes in EPFs' pathogenesis have been extensively studied, the precise ultrastructural mechanism of the fungal infection, particularly penetration of the host insect cuticles, is not well understood.RESULTS: In this study, we investigated the morphology and ultrastructure of the larval cuticle of the red flour beetle, Tribolium castaneum, after treatment with B. bassiana ERL1170 expressing an enhanced green fluorescent protein (Bb-eGFP). The Bb-eGFP showed high virulence against the larvae, with approximately 90% mortality at 48 h after treatment (HAT) and 100% at 72 HAT under our infection conditions. In these larvae, the regions of the body wall with flexible cuticles, such as the ventral and ventrolateral thorax and abdomen, became darkly melanized, but there was little to no melanization in the rigid dorsal cuticular structures. Confocal microscopy and transmission electron microscopy (TEM) indicated that germinated conidia on the surface of the larval cuticle were evident at 6 HAT, which formed penetration pegs and began to penetrate the several cuticle layers/laminae by 12 HAT. The penetration pegs then developed invading hyphae, some of which passed through the cuticle and reached the epidermal cells by 24 HAT. The larval cuticle was aggressively and extensively disrupted by 48 HAT, and a number of outgrowing hyphae were observed at 72 HAT. CONCLUSIONS: Our results indicate that Bb-eGFP is capable of infection and penetrating T. castaneum larvae shortly after inoculation (∼24 HAT) at the body regions with apparently flexible and membranous cuticles, such as the ventral intersegmental regions and the ventrolateral pleura. This study provides details on the histopathogenesis of the host cuticle by infection and penetration of EPFs, which can facilitate the management of insect pests.
Monochamus alternatus is a major forest pest that spreads pine wilt disease in pine trees as a vector of pine wilt nematodes. Chemical insecticides used as fumigants to control overwintering M. alternatus in forests are highly toxic to the environment, so we investigated entomopathogenic fungus Beauveria bassiana ERL836 as an eco-friendly and alternative material to control overwintering M. alternatus. In this work, we evaluated the insecticidal activity of B. bassiana ERL836 against M. alternatus adults, the possibility of fungal colonization on pine tree bark, and finally the control efficacy of fungal pre-treatment on pine tree logs against emerging M. alternatus adults in semi-field and field conditions. M. alternatus adults were killed on the pine tree logs pre-treated with the B. bassiana ERL836. White conidia were observed not only on the surface of the dead adults but also on the pine tree logs, suggesting that the adults were killed by the fungus on the pine. A formulated ERL836 powder treatment on larvae-infested pine logs showed high insecticidal activity against adults, similar to that with the fungal powder suspension treatment, but we demonstrated that using the fungal powder was simpler than using the suspension in field conditions. Even in the field condition, the fungal powder treatment showed high insecticidal activity against M. alternatus adults, which we attribute to its ability to maintain fungal activity for a long time in field conditions by covering the pine tree logs with a film during overwintering. We confirmed that the risk that fungus-infected M. alternatus adults would spread the fungus to other non-target forest insects was low. Thus, even a high-concentration treatment in a specific area is unlikely to transmit the fungus outside that area, so it can be safely used to control this pine wilt nematode vector in forest ecosystems.
In order to establish a strategy for controlling pests living in trees and causing damage, injected entomopathogenic fungal conidia suspension of Beauveria bassiana into a four‐year‐old Canary Island date palm, Phoenix canariensis. As a result, B. bassiana Bb‐egfp#3 expressing green fluorescent protein was dispersed from the injection site to the top and bottom of the tree. To evaluate the insecticidal activity of the Bb‐egfp#3 injected into the tree, we released mealworm larvae (Tenebrio molitor) on the ground palm tree sample that was injected with the conidia suspension. As a result, mycosis caused by the injected Bb‐egfp#3 in the trees was generated in all cadavers. This study suggests the possibility of establishing a new control strategy against tree pests by injecting an entomopathogenic fungal conidia suspension into trees.
The entomopathogenic fungus Beauveria bassiana JEF-350 was isolated from forest soil in South Korea. Here, we report the whole-genome sequence of JEF-350, along with the analyzed genetic information, which can be used to study insecticidal mechanisms and fungal diversity.
Background: Insect-killing fungal species, Beauveria bassiana, is as an environment-friendly pest management tool, and many isolates are on the track of industrialization. However, some of B. bassiana isolates show similar morphology and virulence against insect pests, and so it is hard to differentiate them. Herein we used two patented isolates, ERL836 and JEF-007, and investigated their virulence against western flower thrips, Frankliniella occidentalis, and further analyzed genome structures and transcriptional responses when infecting the thrips to see possible differences.Results: The two isolates showed no significant differences in fungal growth, conidial production, virulence against thrips, and they were structurally similar in genome. But, in transcription level, ERL836 appeared to infect thrips easily, while JEF-007 appeared to have more difficulty. In the GO analysis of ERL836 DEGs (differentially expressed genes), the number of up-regulated genes was much larger than that of down-regulated genes, when compared to JEF-007 DEGs (more genes down-regulated). Interestingly, in the enrichment analysis using shared DEGs between two infecting isolates, plasma membrane-mediated transporter activity and fatty acid degradation pathway including cytochrome P450 were more active in infecting ERL836.Conclusion: The two B. bassiana isolates had similar morphology and virulence as well as genome structure, but in transcription level they differently infected western flower thrips. This comparative approach using shared DEG analysis could be easily applied to characterize the difference of the two B. bassiana isolates, JEF-007 and ERL836.
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