Plant‑herbivorous insects are a severe danger to the world’s agricultural production of various crops. Insecticides used indiscriminately resulted in habitat destruction due to their high toxicity, as well as disease resist‑ ance. In this respect, the development of a sustainable approach to supreme crop production with the least damage is a crucially prerequisite. As a result, the current study was carried out to understand the potential effect of arbuscu‑ lar mycorrhizal (AM) fungi along with Beauvaria bassiana silica nanoparticles (Si NPs) as a new approach to increase cotton (Gossypium hirsutum L. Merr.) defense against an insect herbivore, Spodoptera littoralis. AM and non‑AM cotton plants were infested with S. littoralis and then sprayed with a biopesticide [B. bassiana Si NPs] or a chemical insecticide (Chlorpyrifos). Results: The gas chromatography‑mass spectrometry (GC–MS) analysis of B. bassiana Si NPs fungal extract showed that the major constituents identified were Oleyl alcohol, trifluoroacetate, 11‑Dodecen‑1‑AL and 13‑Octadecenal, (Z)‑(CAS). Besides, results revealed a highly significant decrease in growth parameters in S. littoralis infested plants, however, with AM fungal inoculation a substantial improvement in growth traits and biochemical parameters such as protein and carbohydrates contents was observed. In addition, stimulation in proline and antioxidant enzymes activ‑ ity and a decrease in malondialdehyde content were observed after AM inoculation. Conclusion: AM fungi mitigate the harmful effects of herbivorous insects by strengthening the cotton plant’s health via enhancing both morphological and biochemical traits that can partially or completely replace the application of chemical insecticides.