Most assassin bugs are predators that act as important natural enemies of insect pests. Mitochondrial (mt) genomes of these insects are double-strand circular DNAs that encode 37 genes. In the present study, we explore the duplication and rearrangement of tRNA genes in the mt genome of Reduvius tenebrosus, the first mt genome from the subfamily Reduviinae. The gene order rearranges from CR (control region)-trnI-trnQ-trnM-ND2 to CR-trnQ-trnI2-trnI1-trnM-ND2. We identified 23 tRNA genes, including 22 tRNAs commonly found in insects and an additional trnI (trnI2), which has high sequence similarity to trnM. We found several pseudo genes, such as pseudo-trnI, pseudo-CR, and pseudo-ND2, in the hotspot region of gene rearrangement (between the control region and ND2). These features provided evidence that this novel gene order could be explained by the tandem duplication/random loss (TDRL) model. The tRNA duplication/anticodon mutation mechanism further explains the presence of trnI2, which is remolded from a duplicated trnM in the TDRL process (through an anticodon mutation of CAT to GAT). Our study also raises new questions as to whether the two events proceed simultaneously and if the remolded tRNA gene is fully functional. Significantly, the duplicated tRNA gene in the mitochondrial genome has evolved independently at least two times within assassin bugs.

Asian citrus psyllid (ACP), Diaphorina citri (Hemiptera: Liviidae), is one of the most devastating pests in citrus orchards due to its role in transmitting Huanglongbing (HLB). Currently, chemical control remains the most effective strategy for ACP management. Mineral oils are commonly used as insecticides or adjuvants in integrated pest management (IPM) practices. To extend the product life of synthetic pesticides, we evaluated the toxicity of chlorpyrifos (CPF), thiamethoxam (THX), or pyriproxyfen (PPF) mixed with mineral oil Lvying (LY) against ACP nymphs under different weather conditions. Individual application of CPF, THX, and PPF effectively controlled against ACP nymphs under no rain conditions, with mortality rates varying from 20 to 100% during 1–5 d after treatment. The addition of LY at 1.0% or 0.5% rates to CPF, THX, and PPF significantly enhanced their control efficacy, achieving sustained mortality rates from 60 to 100% during the same period. Light rain had less impact on the control efficacy of these insecticide mixtures compared to individual insecticides. However, cumulative rainfall above 20 mm significantly reduced the control efficacy of individual insecticides and their mixtures. The addition of LY decreased both surface tension and contact angle of THX solution on citrus leaves, thereby enhancing the solution’s wetting and increasing THX deposition. Moreover, the rainfastness of THX was improved after adding LY, leading to a greater retention on the leaves. LY at a rate of 1.0% exhibited excellent efficacy against ACP nymphs, and observations using scanning electron microscopy (SEM) showed that LY altered ACP mouthpart morphology and blocked spiracles, likely contributing to its insecticidal effects. This study revealed that mineral oils can serve as both insecticides to combine with synthetic pesticides for enhancing toxicity against ACP and as adjuvants to facilitate the deposition and rainfastness of synthetic pesticides on leaves, which could be recommended for sustainable ACP management in citrus orchards.