Dang Shao-Xia, Zhu Run-Qi, Wang Shu-Jing, Chen Ju-Hong, Ren Yi-Peng, Bu Wen-Jun, Xue Huai-Jun
Institute of Entomology, College of Life Sciences, Nankai University, Tianjin, China.
Pest Manag Sci. 2025 Sep;81(9):5903-5916. doi: 10.1002/ps.8944. Epub 2025 Jun 2.
Laccase2 is a member of the multicopper oxidase family that plays essential roles in the sclerotization and pigmentation of the insect cuticle. Previous studies have suggested RNA interference (RNAi) targeting laccase2 often resulted in high mortality in various pests. This study aims to evaluate the potential effects of laccase2 gene knockdown on a serious pest, Halyomorpha halys (Stål) (Hemiptera: Pentatomidae).
We identified, validated and characterized one laccase2 (Hhlac2) gene from H. halys. Quantitative real-time PCR results demonstrated that the expression of the Hhlac2 is tissue-specific and changes periodically during development. The results of RNA interference revealed that injecting nymphs with the double-stranded RNA Hhlac2 (dsHhlac2) at doses ranging from 100 to 500 ng/individual significantly suppressed Hhlac2 expression, and pigmentation was not significantly inhibited until the injection dose reached more than 500 ng/individual, while the mortality rate reached over 90% at a dose of 100 ng. RNA-sequencing analysis revealed 802 up-regulated and 1052 down-regulated genes in the dsHhlac2-injected group compared with control, and these unigenes were mainly enriched in terms of tyrosine metabolism, melanin biosynthesis, chitin, immunity and apoptosis. Notably, several key differentially expressed genes associated with metabolism (tyrosine metabolism, lipid metabolism and chitin metabolism) and innate immunity (Toll signaling pathway) were mainly significantly suppressed, which may influence host fitness together with phenotypic changes and increased mortality.
Our study elucidates the pivotal roles of Hhlac2 knockdown induces overall disorder and high mortality at the molecular and individual levels, respectively, which provide insight into biocontrol via RNAi-based technology in heteropteran pests. © 2025 Society of Chemical Industry.
漆酶2是多铜氧化酶家族的成员，在昆虫表皮的硬化和色素沉着中起重要作用。先前的研究表明，靶向漆酶2的RNA干扰（RNAi）通常会导致多种害虫的高死亡率。本研究旨在评估敲低漆酶2基因对一种严重害虫——褐飞虱（Halyomorpha halys (Stål)，半翅目：蝽科）的潜在影响。
我们从褐飞虱中鉴定、验证并表征了一个漆酶2（Hhlac2）基因。定量实时PCR结果表明，Hhlac2的表达具有组织特异性，并且在发育过程中周期性变化。RNA干扰结果显示，以100至500 ng/个体的剂量向若虫注射双链RNA Hhlac2（dsHhlac2）可显著抑制Hhlac2的表达，直到注射剂量超过500 ng/个体时色素沉着才受到显著抑制，而在100 ng的剂量下死亡率达到90%以上。RNA测序分析显示，与对照组相比，注射dsHhlac2的组中有802个基因上调，1052个基因下调，这些单基因主要富集在酪氨酸代谢、黑色素生物合成、几丁质、免疫和凋亡方面。值得注意的是，几个与代谢（酪氨酸代谢、脂质代谢和几丁质代谢）和先天免疫（Toll信号通路）相关的关键差异表达基因主要受到显著抑制，这可能与表型变化和死亡率增加一起影响宿主适应性。
我们的研究阐明了敲低Hhlac2分别在分子和个体水平上诱导整体紊乱和高死亡率的关键作用，这为通过基于RNAi的技术对异翅目害虫进行生物防治提供了见解。© 2025化学工业协会。
