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The use of beneficial fungi and bacteria stimulate plant growth and serve to improve yield and food quality in a sustainable manner. The electrical conductivity of nutrients solution is closely linked to better nutrition of vegetable plants in a hydroponic system. Therefore, objectives of current study were to evaluate the effect of isolated and combined inoculation with Azospirillum brasilense and Trichoderma harzianum under two electrical conductivities on growth, nutrition, and yield of lettuce in hydroponic cultivation. The experiment was designed in a strip-plot block with five replications in a 4 × 2 factorial scheme. The treatments were consisted of four microbial inoculations (without, A. brasilense, T. harzianum and co-inoculation) and electrical conductivities (1.2 and 1.4 dS m -1 ). Inoculation with A. brasilense and T. harzianum increased lettuce root growth by 47% and 20%, respectively. The single inoculation of T. harzianum provided higher fresh leaves yield (24%) at electrical conductivity of 1.2 dS m -1 , while single inoculation with A. brasilense increased fresh leaves yield by 17% at electrical conductivity 1.4 dS m -1 . The lowest shoot NO 3 - accumulation (40%) was observed with inoculation of A. brasilense and highest (28%) with inoculation T. harzianum in both electrical conductivities. Inoculation with A. brasilense increased leaf accumulation of K, P, Ca, Mg, Fe, Mn, Cu, and Zn, which are essential for human nutrition and being recommended to improve yield of lettuce plants in hydroponics. It is recommended to use EC 1.4 dS m -1 of the nutrients solution to improve accumulation of K, Mn, Cu, and Zn, regardless of inoculations for biofortification of lettuce with application of fertilizers., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)