Department of Pathogen Biology, Institute of Tropical Medicine, School of Public Health, Southern Medical University, Guangzhou 510515, China.
College of Life Sciences, South China Agricultural University, Guangzhou 510515, China.
Toxins (Basel). 2022 Feb 17;14(2):147. doi: 10.3390/toxins14020147.
Mosquito densoviruses (MDVs) are mosquito-specific viruses that are recommended as mosquito bio-control agents. The MDV densovirus (AeDNV) is a good candidate for controlling mosquitoes. However, the slow activity restricts their widespread use for vector control. In this study, we introduced the () toxin Cry11Aa domain II loop α8 and Cyt1Aa loop β6-αE peptides into the AeDNV genome to improve its mosquitocidal efficiency; protein expression was confirmed using nanoscale liquid chromatography coupled to tandem mass spectrometry (nano LC-MS/MS). Recombinant plasmids were transfected into mosquito C6/36 cell lines, and the expression of specific peptides was detected through RT-PCR. A toxicity bioassay against the first instar larvae revealed that the pathogenic activity of recombinant AeDNV was significantly higher and faster than the wild-type () viruses, and mortality increased in a dose-dependent manner. The recombinant viruses were genetically stable and displayed growth phenotype and virus proliferation ability, similar to wild-type AeDNV. Our novel results offer further insights by combining two mosquitocidal pathogens to improve viral toxicity for mosquito control.
蚊虫浓核病毒（MDVs）是一类专性感染蚊虫的病毒，被推荐作为蚊虫生物防治剂。蚊虫浓核病毒（AeDNV）是控制蚊虫的候选者。然而，其缓慢的杀虫活性限制了其在病媒控制中的广泛应用。在本研究中，我们将Cry11Aa 毒素的 II 环α8 和 Cyt1Aa 的 loop β6-αE 肽引入 AeDNV 基因组中，以提高其杀蚊效率；使用纳升液相色谱-串联质谱法（nano LC-MS/MS）进行蛋白表达验证。将重组质粒转染到蚊虫 C6/36 细胞系中，通过 RT-PCR 检测特异性肽的表达。对第一龄幼虫进行毒性生物测定表明，重组 AeDNV 的致病活性明显高于野生型（wt）病毒，死亡率呈剂量依赖性增加。重组病毒具有遗传稳定性，表现出与野生型 AeDNV 相似的生长表型和病毒增殖能力。我们的新结果通过结合两种杀蚊病原体来提高病毒毒性以控制蚊虫，提供了进一步的见解。
