Diisononyl phthalate (DINP) as a phthalate plasticizer is widely used in daily life and production, which shows endocrine disruption effects and has several adverse effects on the normal physiological function. Here, the effects of DINP (0.1%, 0.2%, 0.5%, and 1.0%) (v/v) on the fertility, lifespan, climbing behavior, anti-starvation ability of Drosophila melanogaster and the potential modes of action were investigated. The results showed that DINP impaired fertility in a dose-dependent manner and smaller ovarian volume, lower hatching rate, and fewer offspring was observed at higher concentrations. The effect of DINP on the lifespan showed gender-specific, and mortality was increased after exposure above 0.2% DINP. The climbing ability increased at 0.1% DINP compared with the vehicle group, while it manifested a dose-dependent decrease at higher concentrations. The anti-starvation ability exhibited hormesis after short-term culture and reduced as culture time extending. By measuring the redox status (catalase (CAT) and reactive oxygen species (ROS)) of adult flies after two exposure methods, it was found that DINP induced redox instability, which may explain the above effects at the molecular level. This study provides data to support a comprehensive analysis of DINP potential toxicity and to guide its rational use and management better.Copyright © 2020. Published by Elsevier Inc.