The toxic effects of nanoplastics on transgenerational toxicity in environmental organisms and the involved mechanisms remain poorly comprehended. This study aimed to identify the role of SKN-1/Nrf2-dependent regulation of mitochondrial homeostasis in response to transgenerational toxicity caused by changes in nanoplastic surface charges in Caenorhabditis elegans (C. elegans). Our results revealed that compared with the wild-type control and PS exposed groups, exposure to PS-NH<sub>2</sub> or PS-SOOOH at environmentally relevant concentrations (ERC) of &#
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xa0;1&#
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x3bc;g/L caused transgenerational reproductive toxicity, inhibited mitochondrial unfolded protein responses (UPR) by downregulating the transcription levels of
hsp-6,
ubl-5,
dve-1,
atfs-1,
haf-1, and
clpp-1, membrane potential by downregulating
phb-1 and
phb-2, and promoted mitochondrial apoptosis by downregulating
ced-4 and
ced-3 and upregulating
ced-9, DNA damage by upregulating
hus-1,
cep-1,
egl-1, reactive oxygen species (ROS) by upregulating
nduf-7 and
nuo-6, ultimately resulting in mitochondrial homeostasis. Additionally, further study indicated that SKN-1/Nrf2 mediated antioxidant response to alleviate PS-induced toxicity in the P0 generation and dysregulated mitochondrial homeostasis to enhance PS-NH<sub>2</sub> or PS-SOOOH-induced transgenerational toxicity. Our study highlights the momentous role of SKN-1/Nrf2 mediated mitochondrial homeostasis in the response to nanoplastics caused transgenerational toxicity in environmental organisms.