Department of Pathogen Biology, Guangdong Provincial Key Laboratory of Tropical Disease Research, Southern Medical University, Guangzhou, Guangdong Province, China.
PLoS Negl Trop Dis. 2019 Jul 15;13(7):e0007590. doi: 10.1371/journal.pntd.0007590. eCollection 2019 Jul.
The entomopathogenic fungus Beauveria bassiana has been widely used to kill mosquito larvae and adults in the laboratory and field. However, its slow action of killing has hampered its widespread application. In our study, the B. bassiana fungus was genetically modified to express the Bacillus thuringiensis (Bt) toxin Cyt2Ba to improve its efficacy in killing mosquitoes.
METHODOLOGY/PRINCIPAL FINDINGS: The efficacy of the wild type (WT) of B. bassiana and a transgenic strain expressing Cyt2Ba toxin (Bb-Cyt2Ba) was evaluated against larval and adult Aedes mosquitoes (Aedes aegypti and Aedes albopictus) using insect bioassays. The Bb-Cyt2Ba displayed increased virulence against larval and adult Aedes mosquitoes compared with the WT: for Ae. aegypti adults, the median lethal time (LT50) was decreased by 33% at the concentration of 1× 108 conidia/ml, 19% at 1× 107 conidia/ml and 47% at 1× 106 conidia/ml. The LT50 for Ae. albopictus adults was reduced by 20%, 23% and 29% at the same concentrations, respectively. The LT50 for Ae. aegypti larvae was decreased by 42% at 1× 107 conidia/ml and 25% at 1× 106 conidia/ml, and that for Ae. albopictus larvae was reduced by 33% and 31% at the same concentrations, respectively. In addition, infection with Bb-Cyt2Ba resulted in a dramatic reduction in the fecundity of Aedes mosquitoes.
CONCLUSIONS/SIGNIFICANCE: In conclusion, our study demonstrated that the virulence of B. bassiana against mosquitoes can be significantly improved by introducing the Bt toxin gene Cyt2Ba into the genome to express the exogenous toxin in the fungus. The transgenic strain Bb-Cyt2Ba significantly reduced the survival and fecundity of Ae. aegypti and Ae. albopictus compared with the WT strain, which suggested that this recombinant B. bassiana has great potential for use in mosquito control.
球孢白僵菌已被广泛用于实验室和野外杀死蚊幼虫和成虫。然而,其缓慢的杀虫作用阻碍了其广泛应用。在我们的研究中,通过基因修饰球孢白僵菌表达苏云金芽孢杆菌(Bt)毒素 Cyt2Ba 来提高其杀蚊效果。
方法/主要发现:采用昆虫生物测定法评价野生型(WT)球孢白僵菌和表达 Cyt2Ba 毒素的转基因菌株(Bb-Cyt2Ba)对幼虫和成虫埃及伊蚊(Aedes aegypti 和 Aedes albopictus)的效果。与 WT 相比,Bb-Cyt2Ba 对埃及伊蚊幼虫和成虫表现出更高的毒力:在 1×108 个分生孢子/ml 的浓度下,对埃及伊蚊成蚊的半数致死时间(LT50)降低了 33%,在 1×107 个分生孢子/ml 的浓度下降低了 19%,在 1×106 个分生孢子/ml 的浓度下降低了 47%。在相同浓度下,对致倦库蚊成蚊 LT50 分别降低了 20%、23%和 29%。在 1×107 个分生孢子/ml 的浓度下,对埃及伊蚊幼虫 LT50 降低了 42%,在 1×106 个分生孢子/ml 的浓度下降低了 25%,在相同浓度下,对致倦库蚊幼虫 LT50 降低了 33%和 31%。此外,Bb-Cyt2Ba 感染导致埃及伊蚊和致倦库蚊的繁殖力显著降低。
总之,本研究表明,通过将 Bt 毒素基因 Cyt2Ba 引入基因组,在真菌中表达外源毒素,可显著提高球孢白僵菌对蚊子的毒力。与 WT 菌株相比,转基因菌株 Bb-Cyt2Ba 显著降低了埃及伊蚊和致倦库蚊的存活率和繁殖力,这表明这种重组球孢白僵菌在控制蚊子方面具有很大的潜力。