Chen Xuanhao, Yao Shuyuan, Xie Liangqian, Li Jinyang, Xiong Lei, Yang Xiaozhen, Chen Yi, Cao Fang, Hou Qing, You Minsheng, Liu Yuanyuan, Gurr Geoff M, You Shijun
State Key Laboratory of Ecological Pest Control for Fujian and Taiwan Crops, College of Life Science, Haixia Institute of Science and Technology, Fujian Agriculture and Forestry University, Fuzhou, 350002, China; Ministerial and Provincial Joint Innovation Centre for Safety Production of Cross-Strait Crops, Fujian Agriculture and Forestry University, Fuzhou, 350002, China; Joint International Research Laboratory of Ecological Pest Control, Ministry of Education, Fuzhou, 350002, China.
Institute of Plant Protection, Jiangxi Academy of Agricultural Sciences, Nanchang, 330200, China.
Insect Biochem Mol Biol. 2025 Feb;177:104248. doi: 10.1016/j.ibmb.2024.104248. Epub 2024 Dec 12.
The olfactory system of insects plays a pivotal role in multiple, essential activities including feeding, mating, egg laying, and host localization. The capacity of odorant receptors to recognize odor molecules relies on odorant receptor co-receptors forming heterodimers. Here we report the successful engineering a homozygous mutant strain of diamondback moth (Plutella xylostella) in which the odorant receptor co-receptor PxOrco was silenced using CRISPR/Cas9. This insect is a globally important crop pest for which novel control methods are urgently required. Behavioral assays demonstrated that PxOrco knockout males exhibited abolished courtship behaviors, inability to mate, and loss of selective preference for P. xylostella's key sex pheromone components. Whilst female mating behavior and fecundity remained unaffected by PxOrco knockout, oviposition response to leaf alcohol, a key cue for normal oviposition behavior, was lost. Electroantennography revealed drastically reduced responses to sex pheromones and plant volatiles in PxOrco-deficient adults but food location by larvae was unaffected. Moreover, expression analysis of PxOrco-deficient pheromone receptors (PRs) indicated varied regulation patterns, with down-regulation observed in several PRs in both sexes. These findings underscore the critical role of PxOrco in regulating multiple olfactory aspects in P. xylostella, including feeding, mating, and host location. Our study identifies the potential of disrupting the Orco gene in this and other pest species to provide novel avenues for future pest control.
昆虫的嗅觉系统在多种重要活动中起着关键作用，这些活动包括取食、交配、产卵和宿主定位。气味受体识别气味分子的能力依赖于气味受体共受体形成异二聚体。在此，我们报告成功构建了小菜蛾（Plutella xylostella）的纯合突变品系，其中利用CRISPR/Cas9使气味受体共受体PxOrco沉默。这种昆虫是一种全球重要的农作物害虫，迫切需要新的防治方法。行为分析表明，PxOrco基因敲除的雄虫求偶行为消失、无法交配，并且对小菜蛾关键性信息素成分丧失了选择性偏好。虽然PxOrco基因敲除对雌虫的交配行为和繁殖力没有影响，但对正常产卵行为的关键线索叶醇的产卵反应丧失。触角电位图显示，PxOrco缺陷型成虫对性信息素和植物挥发物的反应大幅降低，但幼虫的食物定位不受影响。此外，对PxOrco缺陷型性信息素受体（PRs）的表达分析表明其调控模式各异，在雌雄两性的几个PRs中均观察到下调。这些发现强调了PxOrco在调节小菜蛾包括取食、交配和宿主定位在内的多个嗅觉方面的关键作用。我们的研究确定了在该害虫及其他害虫物种中破坏Orco基因的潜力，为未来害虫防治提供了新途径。
