This experiment was carried out during the two successive seasons of 2019/2020 and 2020/2021 at El-Kasasin Research Station. The current study was conducted to determine the effect of bio-fertilizer and some microelements on the growth, yield, and active ingredients of Plantago psyllium plants. This experiment included five treatments, as following: Control, bio-fertilizer + (Zn and Mn), bio-fertilizer + (Fe and Mn), bi-ofertilizer + (Fe and Zn) and bio-fertilizer + (Fe, Mn, and Zn). The treatments were arranged in a complete randomized block design with three replicates. The best treatment was bio-fertilizer + (Fe, Mn, and Zn), which recorded the highest values in vegetative growth including plant height (40.41 and 41.68 cm) and number of branches/plant (22.67 and 24.33), in the first and second seasons, respectively. Also, there was a significant increase in seed yield; seed weight per plant was 8.23 g and 8.57 g, and the seed weight per feddan was 276.64 kg and 287.84 kg,  in the first and second seasons, respectively, when using the same treatment. Furthermore, the results showed that an increase in oil yield; fixed oil percentage of 15.64% and 15.95%, oil yield per plant of 1.29 ml and 1.37 ml, and oil yield per feddan of 43.27l and 45.91l when treated with bio-fertilizer  + (Fe, Mn, and Zn) and gave a significant increase in the active ingredients (mucilage of 30.20% and 30.26%, protein of 22.33% and 22.83%, seed husk/plant of 27.60 g and 27.72 g and carbohydrates of 49.87% and 51.23% when using the same treatment, in the first and second seasons, respectively.