Global climate change is characterized by increased extreme temperatures affecting insects at all trophic levels. Zinc finger proteins (ZFPs) are key regulators of gene expression and cell differentiation in eukaryotes, essential for stress resistance in both animals and plants. Using CRISPR/Cas9 for gene deletion, this study predicted and examined the structure of ZFP320 in the diamondback moth (Plutella xylostella) and investigated its function in temperature stress response through a comprehensive age-stage, two-sex life table analysis. We found ZFP320 encodes a 387 amino acid protein (43 kDa) with no transmembrane domains, featuring a ZnF-C2H2 domain. Quantitative fluorescence analysis showed that ZFP320 expression increased under high temperatures. ZFP320 knockout altered antioxidant gene expression, resulting in higher levels of superoxide dismutase and catalase in mutant strains compared with wild-type strain. Life table analysis revealed that the mutant strains had shorter fecundity and oviposition periods under both normal and high temperatures. Additionally, mutant strains exhibited lower parameters (r, λ, R₀), as well as reduced survival rates and critical thermal maxima. Notably, PxZFP320 plays a crucial role in temperature adaptation, paving the way for future investigations on the significance of ZFPs in P. xylostella's temperature tolerance.

Keywords: CRISPR/Cas9; Plutella xylostella; age-stage two-sex life table; temperature adaptability.

全球气候变化的特点是极端温度增加，影响了所有营养级的昆虫。锌指蛋白（ZFPs）在真核生物中是基因表达和细胞分化的关键调节因子，在动植物中的应激抵抗中必不可少。本研究使用CRISPR/Cas9进行基因敲除技术，预测并分析了小菜蛾(Plutella xylostella)中的ZFP320结构，并通过全面的年龄阶段、两性生命表分析来调查其在温度压力反应中的功能。我们发现ZFP320编码一个由387个氨基酸组成的蛋白质（43 kDa），不包含跨膜域，具有ZnF-C2H2结构域。定量荧光分析显示，在高温条件下ZFP320的表达量增加。ZFP320敲除改变了抗氧化基因的表达，在突变株中导致了超氧化物歧化酶和过氧化氢酶水平高于野生型株。生命表分析表明，无论是在正常温度还是在高温下，突变株的繁殖期和产卵期都比野生型短。此外，突变株还表现出较低的参数（r、λ、R₀），以及更低的生存率和临界最高温度。值得注意的是，PxZFP320在温度适应中起着关键作用，为未来研究小菜蛾P. xylostella温度耐受性的锌指蛋白的重要性铺平了道路。

关键词：CRISPR/Cas9；Plutella xylostella；年龄阶段两性生命表；温度适应性。