Xie Jingang, Ali Amjad, Li Yuan, Zhuang Ziyan, Liu Xiaoning
Xinjiang Key Laboratory of Biological Resources and Genetic Engineering, College of Life Science and Technology, Xinjiang University, Urumqi, China.
J Econ Entomol. 2025 Feb 11;118(1):381-390. doi: 10.1093/jee/toae283.
Tuta absoluta has developed resistance to many biological insecticides, causing substantial agricultural and economic losses annually. P450s have been the most extensively studied enzymes in the context of insecticide metabolism in insect pests, and the detoxification metabolism of P450s in T. absoluta against biological insecticides remains poorly understood. In T. absoluta, CYP304F1 was screened from the comparative transcriptome of 2 regional populations in Xinjiang, China. The objective of the present study was to characterize and analyze CYP304F1 of T. absoluta and explore its role in detoxification of spinetoram as well as the growth and development of T. absoluta. Following cloning and sequence analysis of the target gene, it was named CYP304F1. Expression levels of CYP304F1 were then determined after spinetoram exposure and across various developmental instars and tissues. Finally, dsCYP304F1 was synthesized and utilized to assess the effects of post-RNAi on larval spinetoram susceptibility, growth, and development. Sequence analysis revealed that CYP304F1 harbors conserved domains characteristic of P450 proteins, exhibiting high conservation within the Lepidoptera clade. Treatment with an LC50 dose of spinetoram significantly upregulated CYP304F1 expression in T. absoluta larvae. Silencing CYP304F1 significantly enhanced larval susceptibility to spinetoram and prolonged leaf-mining duration and developmental time from the 2nd instar to 4th instar by 40% and 17.6%, respectively, compared to controls. And feeding on dsCYP304F1-treated leaves for 6 days resulted in 71% larval mortality. These results suggested that CYP304F1 played a crucial role in detoxification of spinetoram as well as in the growth and development of T. absoluta larvae.
番茄潜叶蛾已对多种生物杀虫剂产生抗性，每年造成巨大的农业和经济损失。在害虫杀虫剂代谢方面，细胞色素P450（P450s）是研究最为广泛的酶，而番茄潜叶蛾中P450s对生物杀虫剂的解毒代谢仍知之甚少。在中国新疆的两个区域种群的比较转录组中筛选出了番茄潜叶蛾的CYP304F1。本研究的目的是对番茄潜叶蛾的CYP304F1进行表征和分析，探索其在多杀菌素解毒以及番茄潜叶蛾生长发育中的作用。对目标基因进行克隆和序列分析后，将其命名为CYP304F1。然后在多杀菌素处理后以及不同发育龄期和组织中测定CYP304F1的表达水平。最后，合成双链RNA（dsCYP304F1）并用于评估RNA干扰后对幼虫多杀菌素敏感性、生长和发育的影响。序列分析表明，CYP304F1具有P450蛋白的保守结构域，在鳞翅目进化枝中表现出高度保守性。用多杀菌素的半数致死浓度（LC50）剂量处理显著上调了番茄潜叶蛾幼虫中CYP304F1的表达。与对照相比，沉默CYP304F1显著增强了幼虫对多杀菌素的敏感性，从二龄到四龄的潜道持续时间和发育时间分别延长了40%和17.6%。取食dsCYP304F1处理的叶片6天导致71%的幼虫死亡。这些结果表明，CYP304F1在多杀菌素解毒以及番茄潜叶蛾幼虫的生长发育中起关键作用。
