AbstractVarious endophytic fungi of native plants and crops are important entomopathogens. The objective of this study was to investigate the entomopathogenic action of Beauveria bassiana (Balsamo) Vuillemin, Metarhizium robertsii (Metchnikoff) Sorokin, and Isaria fumosorosea (Wize) Brown & Smith (all Ascomycota: Hypocreales) against larvae of Sesamia nonagrioides (Lefebvre) (Lepidoptera: Noctuidae) artificially introduced into Sorghum bicolor L. (Moench) (Poaceae) plants under natural environmental conditions. Sorghum bicolor is an economically important crop cultivated for grain, fiber, forage, and lately for biofuel, and S. nonagrioides is its main pest in Mediterranean areas. Young sorghum plants were inoculated with the entomopathogens by spraying in the field. Plant water status, chlorophyll concentration, photosynthesis, and transpiration were not affected. Thirty days after endophyte establishment, plants were infested with fourth instars of S. nonagrioides. The endophytes prevented 50–70% of larvae from entering stalks. Larval mortality was 70–100% and tunnel lengths were reduced by 60–87%. Larval infestation resulted in reduced electron transport capacity and net photosynthetic rate, which was ameliorated in the presence of I. fumosorosea and reversed by B. bassiana and M. robertsii. The growth of sorghum was unaffected in all treatments during the experimental period. Beauveria bassiana and M. robertsii can protect sweet sorghum from damage induced by S. nonagrioides under natural environmental conditions without affecting plant physiology and growth.