College of Agronomy and Biotechnology, Southwest University, Chongqing, China.
Laboratory Animal Center, Southwest University, Chongqing, China.
Pest Manag Sci. 2024 Oct;80(10):4915-4923. doi: 10.1002/ps.8204. Epub 2024 Jun 4.
Entomopathogenic fungi, such as Beauveria bassiana, hold promise as biological control agents against insect pests. However, the efficacy of these fungi can be hindered by insect immune responses. One strategy to enhance fungal virulence is to manipulate host immune by targeting key regulatory molecules like 20-hydroxyecdysone (20E).
In this study, we engineered B. bassiana strains to constitutively express the enzyme ecdysteroid UDP-glucosyltransferase (EGT), which inactivates 20E, a crucial insect molting hormone. The engineered strain Bb::EGT-1 exhibited robust expression of EGT, leading to a significant reduction in insect 20E levels upon infection. Moreover, infection with Bb::EGT-1 resulted in accelerated larval mortality. Immune responses analysis revealed repression of insect immune response genes and decreased phenoloxidase (PO) activity in larvae infected with Bb::EGT-1. Microbiome analysis indicated alterations in bacterial composition within infected insects, with increased abundance observed during infection with Bb::EGT-1. Additionally, the presence of bacteria hindered hyphal emergence from insect cadavers, suggesting a role for microbial competition in fungal dissemination.
Constitutive expression of EGT in B. bassiana enhances fungal virulence by reducing insect 20E levels, suppressing immune responses, and altering the insect microbiome. These findings highlighted the potential of engineered fungi as effective biocontrol agents against insect pests and provide insights into the complex interactions between entomopathogenic fungi, their hosts, and associated microbes. © 2024 Society of Chemical Industry.
拟青霉等虫生真菌有望成为防治害虫的生物防治剂。然而,昆虫的免疫反应可能会阻碍这些真菌的功效。一种增强真菌毒力的策略是通过靶向 20-羟基蜕皮激素(20E)等关键调节分子来操纵宿主免疫。
在这项研究中,我们设计了毕赤酵母菌株,使其组成型表达可使 20E 失活的酶蜕皮甾酮 UDP-葡萄糖基转移酶(EGT),20E 是一种关键的昆虫蜕皮激素。工程菌株 Bb::EGT-1 表现出 EGT 的强表达,导致感染后昆虫 20E 水平显著降低。此外,感染 Bb::EGT-1 导致幼虫死亡率迅速增加。免疫反应分析显示,感染幼虫中昆虫免疫反应基因受到抑制,酚氧化酶(PO)活性降低。微生物组分析表明,感染昆虫体内细菌组成发生变化,感染 Bb::EGT-1 时细菌丰度增加。此外,细菌的存在阻碍了昆虫尸体中菌丝的出现,表明微生物竞争在真菌传播中起作用。
在毕赤酵母中组成型表达 EGT 通过降低昆虫 20E 水平、抑制免疫反应和改变昆虫微生物组来增强真菌毒力。这些发现强调了工程真菌作为防治害虫的有效生物防治剂的潜力,并提供了对昆虫病原真菌与其宿主和相关微生物之间复杂相互作用的深入了解。