Mosquitoes transmit numerous diseases, including malaria and dengue fever, making mosquito control crucial for managing the spread of these and other illnesses. Beauveria bassiana (Balsamo-Crivelli) Vuillemin (Hypocreales: Cordycipitaceae) holds promise for the biological control of mosquito vectors but faces challenges such as low pathogenicity, lengthy infection periods, reduced longevity, and a dependence on specific environmental conditions (e.g., temperature and humidity) for spore germination, thus impacting its efficacy. To address this, a strain of B. bassiana was engineered to carry the SPHvt/GNA fusion pathogenicity protein-encoding gene. A plasmid containing the gpdA promoter, GFP gene, and SPHvt/GNA gene was constructed using molecular cloning techniques, including digestion and ligation, based on the base plasmids pBarGPE1 and pBarGFP. The engineered vector plasmid pBarGFPSPHvt/GNA was integrated into the genome of B. bassiana Bb252 through Agrobacterium tumefaciens-mediated fungal genetic transformation, resulting in B. bassiana strains with the SPHvt/GNA fusion pathogenicity protein-encoding gene. These transgenic strains were identified through fluorescence screening and confirmed at the genomic, transcriptional, and protein levels. A spore suspension of Bb252::BarGFPSPHvt/GNA was applied to Anopheles stephensi Liston (Diptera: Culicidae) larvae. The effect of pathogenicity on larval survival was assessed, and the infection process was monitored. The results indicated that the construction of a transgenic B. bassiana strain for potential management of mosquito larvae.Abstract
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