BACKGROUNDLaccase2 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).RESULTSWe 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.CONCLUSIONOur 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.

中文翻译：

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漆酶 2 对 Halyomorpha halys （Stål） （半翅目：Pentatomidae） 表皮色素沉着和存活影响的综合 RNA 干扰和 RNA 测序分析

背景 Laccase2 是多铜氧化酶家族的一员，在昆虫角质层的硬化和色素沉着中起着重要作用。先前的研究表明，靶向漆酶 2 的 RNA 干扰 （RNAi） 通常会导致各种害虫的高死亡率。本研究旨在评估漆酶 2 基因敲低对严重害虫 Halyomorpha halys （Stål） （半翅目：Pentatomidae） 的潜在影响。结果我们鉴定、验证和表征了 H. halys 的一个漆酶 2 （Hhlac2） 基因。定量实时 PCR 结果表明，Hhlac2 的表达是组织特异性的，并且在发育过程中会周期性地变化。RNA 干扰结果显示，以 100 至 500 ng/个的剂量注射双链 RNA Hhlac2 （dsHhlac2） 显着抑制了 Hhlac2 的表达，直到注射剂量达到 500 ng/个才显着抑制色素沉着，而在 100 ng 剂量下死亡率达到 90% 以上。RNA 测序分析显示，与对照组相比，dsHhlac2 注射组有 802 个上调和 1052 个下调基因，这些单基因主要富集于酪氨酸代谢、黑色素生物合成、几丁质、免疫和细胞凋亡方面。 值得注意的是，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） 相关的几个关键差异表达基因主要受到显著抑制，这可能会影响宿主适应性以及表型变化和死亡率增加。结论我们的研究阐明了 Hhlac2 敲低分别在分子和个体水平上诱导整体疾病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的生物控制提供了见解。© 2025 化工学会.