Allyl isothiocyanate (AITC) is a promising alternative to chemical fumigants, and mitochondrial dysfunction has been proposed to play a crucial role in its lethal mechanisms; however, the specific lethal mechanisms of AITC remain unknown. Four mitochondrial electron transport chain genes, nd5, nd6, cox1, and cox5, were selected from adult Sitophilus zeamais and processed with RNA interference experiments. Then, the biochemical and biophysical effects were compared between double-stranded RNA (dsRNA)-mediated insects and wild-type insects after AITC fumigation at the concentration of LC₅₀ values. The bioactivity of AITC against dsnd6-mediated insects increased, while the bioactivity against dcox1-mediated insects decreased. Compared with the wild-type insects, the increase of reactive oxygen species (ROS) levels by AITC in mitochondria from dsnd6-mediated insects increased by 18.95%, while that of dscox1-mediated insects decreased by 27.45%. The effects of AITC on mRNA expression levels of detoxifying enzymes including CAT (down-regulation effect) and CuZnSOD (overexpression effect) partly recovered in the dsnd5-mediated insects, while a greater effect was observed for dscox1-mediated insects. Molecular docking results indicated that ASN511 at the cox1 subunit was the binding site of AITC by one hydrogen bond, with a bond distance of 2.1 Å. These findings provide insight for further applications of AITC and could provide a novel strategy to investigate lethal mechanisms of insecticides.

异硫氰酸烯丙酯 (AITC) 是一种很有前景的化学熏蒸剂替代品，有人提出线粒体功能障碍在其致死机制中起关键作用；然而，AITC 的具体致死机制仍然未知。四倍线粒体电子传递链的基因，ND5，ND6，COX1，和cox5，从成年选择玉米象，并用RNA干扰实验处理。然后，比较了双链RNA（dsRNA）介导的昆虫和野生型昆虫在LC ₅₀浓度下经过AITC熏蒸后的生化和生物物理效应。值。AITC对dsnd6介导的昆虫的生物活性增加，而对dcox1介导的昆虫的生物活性降低。与野生型昆虫相比，dsnd6介导的昆虫通过AITC增加线粒体中活性氧（ROS）水平增加了18.95%，而dscox1介导的昆虫减少了27.45%。AITC 对解毒酶 mRNA 表达水平的影响，包括 CAT（下调效应）和 CuZnSOD（过表达效应）在 dsnd5 介导的昆虫中部分恢复，而对 dscox1 介导的昆虫观察到更大的影响。分子对接结果表明cox1亚基的ASN511是AITC的一个氢键结合位点，键距为2.1 Å。