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Wheat stem rust causes by Puccinia graminis f. sp. tritici is a serious threat to wheat production all over the world. This study evaluated the efficacy of Bacillus subtilis, B. polymyxa and B. megaterium, Eugenol and leaf extract of Artemisia cina against the disease, and the involved mechanisms in disease resistance. B. subtilis and Eugenol treatments inhibited spore germination and increased incubation and latent periods than other treatments. Additionally, the lowest infection type, pustule length and pustule width, and receptivity (no. of pustules) were recorded in case of B. subtilis and Eugenol treatments compared with the check control. The best results in disease assessment were achieved by foliar spray application 24 h before and after inoculation. Abnormalities, lysis, collapse, and shrinking of urediniospores were recorded in treated leaves. The activities of antioxidant defense enzymes, i.e., catalase (CAT), peroxidase (POX), and polyphenoloxidase (PPO), were significantly increased in B. subtilis and Eugenol treatments. On the other hand, electrolyte leakage was decreased. The active compounds in B. subtilis were revealed by utilizing GC-MS analysis.

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