CRISPR-Cas9-mediated enhancement of Beauveria bassiana virulence with overproduction of oosporein
Abstract Biocontrol agents play a pivotal role in managing pests and contribute to sustainable agriculture. Recent advancements in genetic engineering can facilitate the development of entomopathogenic fungi with desired traits to enhance biocontrol efficacy. In this study, a CRISPR-Cas9 ribonucleop...
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BMC
2024-11-01
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| Series: | Fungal Biology and Biotechnology |
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| Online Access: | https://doi.org/10.1186/s40694-024-00190-5 |
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| author | Gabriel Moura Mascarin Somraj Shrestha Marcio Vinícius de Carvalho Barros Cortes Jose Luis Ramirez Christopher A. Dunlap Jeffrey J. Coleman |
| author_facet | Gabriel Moura Mascarin Somraj Shrestha Marcio Vinícius de Carvalho Barros Cortes Jose Luis Ramirez Christopher A. Dunlap Jeffrey J. Coleman |
| author_sort | Gabriel Moura Mascarin |
| collection | DOAJ |
| description | Abstract Biocontrol agents play a pivotal role in managing pests and contribute to sustainable agriculture. Recent advancements in genetic engineering can facilitate the development of entomopathogenic fungi with desired traits to enhance biocontrol efficacy. In this study, a CRISPR-Cas9 ribonucleoprotein system was utilized to genetically improve the virulence of Beauveria bassiana, a broad-spectrum insect pathogen used in biocontrol of arthropod pests worldwide. CRISPR-Cas9-based disruption of the transcription factor-encoding gene Bbsmr1 led to derepression of the oosporein biosynthetic gene cluster resulting in overproduction of the red-pigmented dibenzoquinone oosporein involved in host immune evasion, thus increasing fungal virulence. Mutants defective for Bbsmr1 displayed a remarkable enhanced insecticidal activity by reducing lethal times and concentrations, while concomitantly presenting negligible or minor pleiotropic effects. In addition, these mutants displayed faster germination on the insect cuticle which correlated with higher density of free-floating blastospores in the hemolymph and accelerated mortality of the host. These findings emphasize the utility of genetic engineering in developing enhanced fungal biocontrol agents with customized phenotypic traits, and provide an efficient and versatile genetic transformation tool for application in other beneficial entomopathogenic fungi. |
| format | Article |
| id | doaj-art-2af9c5a5dfc14c2285d686d6bd612bc0 |
| institution | OA Journals |
| issn | 2054-3085 |
| language | English |
| publishDate | 2024-11-01 |
| publisher | BMC |
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| series | Fungal Biology and Biotechnology |
| spelling | doaj-art-2af9c5a5dfc14c2285d686d6bd612bc02025-08-20T02:32:49ZengBMCFungal Biology and Biotechnology2054-30852024-11-0111112310.1186/s40694-024-00190-5CRISPR-Cas9-mediated enhancement of Beauveria bassiana virulence with overproduction of oosporeinGabriel Moura Mascarin0Somraj Shrestha1Marcio Vinícius de Carvalho Barros Cortes2Jose Luis Ramirez3Christopher A. Dunlap4Jeffrey J. Coleman5Laboratory of Environmental Microbiology, Embrapa Environment, Brazilian Agricultural Research Corporation, Embrapa EnvironmentDepartment of Entomology and Plant Pathology, Auburn UniversityLaboratory of Agricultural Microbiology, Brazilian Agricultural Research Corporation, Embrapa Rice & BeansCrop Bioprotection Research Unit, National Center for Agricultural Utilization Research, USDACrop Bioprotection Research Unit, National Center for Agricultural Utilization Research, USDADepartment of Entomology and Plant Pathology, Auburn UniversityAbstract Biocontrol agents play a pivotal role in managing pests and contribute to sustainable agriculture. Recent advancements in genetic engineering can facilitate the development of entomopathogenic fungi with desired traits to enhance biocontrol efficacy. In this study, a CRISPR-Cas9 ribonucleoprotein system was utilized to genetically improve the virulence of Beauveria bassiana, a broad-spectrum insect pathogen used in biocontrol of arthropod pests worldwide. CRISPR-Cas9-based disruption of the transcription factor-encoding gene Bbsmr1 led to derepression of the oosporein biosynthetic gene cluster resulting in overproduction of the red-pigmented dibenzoquinone oosporein involved in host immune evasion, thus increasing fungal virulence. Mutants defective for Bbsmr1 displayed a remarkable enhanced insecticidal activity by reducing lethal times and concentrations, while concomitantly presenting negligible or minor pleiotropic effects. In addition, these mutants displayed faster germination on the insect cuticle which correlated with higher density of free-floating blastospores in the hemolymph and accelerated mortality of the host. These findings emphasize the utility of genetic engineering in developing enhanced fungal biocontrol agents with customized phenotypic traits, and provide an efficient and versatile genetic transformation tool for application in other beneficial entomopathogenic fungi.https://doi.org/10.1186/s40694-024-00190-5Secondary metaboliteFilamentous fungiBlastosporesGenome editingRibonucleoproteinHomologous recombination |
| spellingShingle | Gabriel Moura Mascarin Somraj Shrestha Marcio Vinícius de Carvalho Barros Cortes Jose Luis Ramirez Christopher A. Dunlap Jeffrey J. Coleman CRISPR-Cas9-mediated enhancement of Beauveria bassiana virulence with overproduction of oosporein Fungal Biology and Biotechnology Secondary metabolite Filamentous fungi Blastospores Genome editing Ribonucleoprotein Homologous recombination |
| title | CRISPR-Cas9-mediated enhancement of Beauveria bassiana virulence with overproduction of oosporein |
| title_full | CRISPR-Cas9-mediated enhancement of Beauveria bassiana virulence with overproduction of oosporein |
| title_fullStr | CRISPR-Cas9-mediated enhancement of Beauveria bassiana virulence with overproduction of oosporein |
| title_full_unstemmed | CRISPR-Cas9-mediated enhancement of Beauveria bassiana virulence with overproduction of oosporein |
| title_short | CRISPR-Cas9-mediated enhancement of Beauveria bassiana virulence with overproduction of oosporein |
| title_sort | crispr cas9 mediated enhancement of beauveria bassiana virulence with overproduction of oosporein |
| topic | Secondary metabolite Filamentous fungi Blastospores Genome editing Ribonucleoprotein Homologous recombination |
| url | https://doi.org/10.1186/s40694-024-00190-5 |
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