Two field experiments were carried out in a Research Farm in Nubaria, Sugar and Refining Company (NSRC), Behaira government, during the two successive fall seasons of 2015/2016 and 2016/2017 to determine the effect of four bio-fertilizer treatments, and four nitrogen fertilizer levels on yield and its components as well as quality of sugar beet (Beta vulgaris L) cv. Gazel. A split plot design with three replications was used, where the nitrogen fertilizer levels were allocated in the main plots and bio-fertilizer treatments were distributed in the sub plots. The results indicated that increasing nitrogen fertilizer rates significantly increased yield characters, roots, top, biologicaland sugar yields (tons fed^-1), since the highest rates of nitrogen (90 kg N fed^-1) produced the highest roots, top, biological, and sugar yields, for the 1^st and 2^nd seasons, respectively. Increasing nitrogen rate up to (90 kg N fed^-1) significantly increased of some quality parameters, total soluble solids (T.S.S) and sucrose concentration in roots juice, since the maximum T.S.S% and sucrose % was achieved by adding 90 kg N fed^-1, for the two seasons. Moreover, bio-fertilizers treatments, (T.S^®) gave the maximum of roots, top, biological, and sugar yields, in the two seasons and significantly increased of some quality parameters, total soluble solids (T.S.S) and sucrose concentration in roots juice by bio-fertilizer treatments, since the maximum T.S.S% (20.00 and 21.52%) and sucrose % (18.79 and 18.27%) was achieved by adding (T.S^®), for the two seasons of 2015/2016 and 2016/2017, respectively. The interaction between nitrogen rates and bio-fertilizers indicated that the highest all harvested yield, total soluble solids (T.S.S) and sucrose % was obtained by application (T.S^® + 90 kg N fed^-1) in both seasons. So, bio-fertilizer treatments proved a major role in crop production optimization and expected to reduce the pollution of the agricultural environment.