The development of biological pesticides is rapidly becoming an integral aspect
of pest management in sustainable agriculture. This study was conducted to evaluate the
effectiveness of Bacillus halotolerans strain B33 against three common seedborne fungal
pathogens—Fusarium graminearum, Alternaria alternata, and Aspergillus flavus. B33 strain
identity was determined using the 16S rRNA and tuf gene sequences. Commercial wheat,
barley, oat, and rye seeds were artificially infected by fungal isolates and then treated with
B33 overnight culture in Nutrient Broth. The obtained results indicate high efficacy against
F. graminearum (83.55–94.38%) and A. alternata (85.05–96.70%), whereby the highest efficacy
was noted on wheat seed and the lowest was detected on rye seed. On the other hand,
B33 achieved 100% efficacy against A. flavus on barley, rye, and oat seeds, while being
96.24% effective against this pathogen on wheat. Principal component analysis indicated
the highest treatment influence on A. flavus. The effect of all tested treatments on seed
germination was statistically significant compared to the controls, whereby the number of
germinated seeds declined as the seed infection rate increased. B. halotolerans strain B33
effectively managed seedborne fungal pathogens, thereby enhancing seed germination
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