Zeng Mu-Zi, Zhou Wei, Wen Shan-Shan, Wu Hao, Zhang Qing, Fu Kai-Yun, Guo Wen-Chao, Shi Ji-Feng
State Key Laboratory of Resource Insects, College of Sericulture, Textile, and Biomass Sciences, Southwest University, Chongqing 400715, China.
Institute of Plant Protection Xinjiang Academy of Agricultural Sciences/Key Laboratory of Integrated Pest Management on Crops in Northwestern Oasis, Ministry of Agriculture/Xinjiang Key Laboratory of Agricultural Biosafety, Urumqi 830091, China.
Insects. 2024 Aug 19;15(8):623. doi: 10.3390/insects15080623.
The Colorado potato beetle () is a major pest of potato crops. While () genes are essential for insect cuticle formation, their roles in pests like remain unclear. This study aims to identify and characterize genes in and explore their functions in larval development and cuticle integrity. We used genomic and transcriptomic databases to identify -family genes, validated through RT-PCR and RACE. Gene expression was analyzed at various developmental stages and tissues using qRT-PCR. RNA interference (RNAi) and Transmission electron microscopy (TEM) were applied to determine the functional roles of these genes. Four -family genes were identified. Spatio-temporal expression analysis indicated significant gene expression during larval molting and pupal stages, especially in the epidermis. RNAi experiments showed that silencing and ' led to reduced larval weight, cuticle thinning, and increased mortality, while knockdown caused abnormal cuticle thickening and molting disruptions. knockdown increased epicuticle and endocuticle thickness without visible phenotypic changes. The study highlights the essential roles of -family genes in maintaining cuticle structure and integrity, suggesting their potential as targets for RNAi-based pest control.
科罗拉多马铃薯甲虫是马铃薯作物的主要害虫。虽然几丁质合成酶基因对于昆虫表皮形成至关重要，但它们在像科罗拉多马铃薯甲虫这样的害虫中的作用仍不清楚。本研究旨在鉴定和表征科罗拉多马铃薯甲虫中的几丁质合成酶基因，并探索它们在幼虫发育和表皮完整性中的功能。我们使用基因组和转录组数据库来鉴定几丁质合成酶基因家族基因，并通过逆转录聚合酶链反应（RT-PCR）和cDNA末端快速扩增（RACE）进行验证。使用实时定量逆转录聚合酶链反应（qRT-PCR）分析不同发育阶段和组织中的基因表达。应用RNA干扰（RNAi）和透射电子显微镜（TEM）来确定这些基因的功能作用。鉴定出四个几丁质合成酶基因家族基因。时空表达分析表明，在幼虫蜕皮和蛹期，特别是在表皮中，基因表达显著。RNAi实验表明，沉默几丁质合成酶1和几丁质合成酶2导致幼虫体重减轻、表皮变薄和死亡率增加，而敲低几丁质合成酶3导致表皮异常增厚和蜕皮中断。敲低几丁质合成酶4增加了上表皮和内表皮厚度，但没有明显的表型变化。该研究突出了几丁质合成酶基因家族基因在维持表皮结构和完整性方面的重要作用，表明它们作为基于RNAi的害虫防治靶标的潜力。
