This study deals with the toxicological impact of the herbicide anilofos on photosynthesis, respiration, nitrogen assimilation, and antioxidant system in a diazotrophic rice field cyanobacterium Anabaena torulosa. Treatment of anilofos (1.25, 2.5, and 5 mg L−1) affected growth, photosynthetic pigments, photosynthesis, and respiration of the cyanobacterium. Although all the photosynthetic pigments were affected, a maximum effect of the herbicide was observed on phycocyanin (51% reduction) followed by the carotenoids. The effect of the herbicide on photosynthetic pigments resulted in 57% decrease in photosynthetic O2 evolution. Studies on the photochemical activity demonstrated that both photosystems (PS I and PS II) were affected by the herbicide. Decrease in the photosynthesis rate resulted in decreased nitrogen assimilation, as revealed by reduced nitrate (20%) and ammonium (26%) uptake and decreased activities of nitrogenase (63% decrease) and glutamine synthetase (22% decrease). This ultimately resulted in the reduced growth of the organism. Activities of superoxide dismutase, catalase, and peroxidase in the presence of anilofos increased by 1.8–3.5 times over control cultures. Proline content increased by 1.6 times, while the content of ascorbate decreased slightly. These results indicate that the organism was able to tolerate the herbicide stress by activating oxidative stress defense mechanism.