Nanoplastic pollution has become an increasing problem due to over-consumption and degradation in ecosystems. A little is known about ecological toxicity and the potential risks of nanoplastics on plants. To better comprehend the hormetic effects of nanoplastics, the experimental design was conducted on the impacts of polymethyl methacrylate (PMMA) on water status, growth, gas exchange, chlorophyll a fluorescence transient, reactive oxygen species (ROS) content (both content and fluorescence visualization), lipid peroxidation and antioxidant capacity (comparatively between leaves and roots). For this purpose, PMMA (10, 20, 50 and 100 mg L−1) was hydroponically applied to Lactuca sativa for 15 days(d). PMMA exposure resulted a decline in the growth, water content and osmotic potential. As based on assimilation rate (A), stomatal conductance (gs), and intercellular CO2 concentrations (Ci), the decreased stomatal limitation (Ls) and, A/Ci and increased intrinsic mesophyll efficiency proved low carboxylation efficiency showing impaired photosynthesis as a non-stomatal limitation. PMMA toxicity increased the trapping fluxes and absorption with a decrease in electron transport fluxes caused the disruption in reaction centers of photosystems. The leaves and roots had a similar effect against PMMA toxicity, with increased superoxide dismutase (SOD) activity. Although, catalase (CAT) and peroxidase (POX) of leaves increased under 10 mg L−1 PMMA, these defense activities failed to prevent radicals from attacking. Compared to the leaves, the lettuce roots showed an intriguing result for AsA-GSH cycle against PMMA exposure. In the roots, the lowest PMMA application provided the high ascorbate/dehydroascorbate (AsA/DHA), GSH/GSSG and the pool of AsA/glutathione (GSH) and non-suppressed GSH redox state. Also, 10 mg L−1 PMMA helped remove high hydrogen peroxide (H2O2) by both glutathione peroxidase (GPX) and glutathione S-transferase (GST). Since this improvement in the antioxidant system could not be continued in roots after higher applications than 20 mg L−1 PMMA, TBARS (Thiobarbituric acid-reactive substances), indicating the level of lipid peroxidation, and H2O2 increased. Our findings obtained from PMMA-applied lettuce provide new information to advance the tolerance mechanism against nanoplastic pollution.