Plants are influenced by pesticides in terms of various physio-biochemical parameters. This study provides initial evidence on the effect of the insecticide spirotetramat on plant physiological characteristics as a non-target organism. Cucumber plants (Cucumis sativus L.) exposed to spirotetramat were studied 10 days after treatment. There was an increase in the activity of antioxidant enzymes including superoxide dismutase, catalase, guaiacol peroxidase, ascorbate peroxidase, glutathione reductase, and phenylalanine ammonia-lyase. Malondialdehyde, total chlorophyll, hydrogen peroxide contents, and electrolyte leakage index was not affected by spirotetramat. Further biochemical analyses revealed an increase in the content of some free amino acids, as well as sucrose, glucose, and fructose. The amount of salicylic acid and also minerals calcium, manganese, copper, zinc, iron, nitrogen, and magnesium, were elevated in spirotetramat-treated plants. Results have shown that spirotetramat can manipulate cucumber plant physiology through induces biochemical responses that are reflected in changes of antioxidative enzymes, amino acids, soluble carbohydrates, salicylic acid, and mineral elements. The findings of this study provide an insight into the side effects of spirotetramat as a chemical with no specific target site in plants that show no adverse effects on plant health indices. This study focuses on observed physiological changes related to toxicity in plants exposed to hazardous substances present in the environment that can improve our knowledge and understanding of the underlying effects of xenobiotics on plants.