Two field experiments were carried out at the research farm of Nubaryia Sugar and Refining Company (NSRC), located at (30°63' 88.93" N latitude; 30°22′ 46.21′′ E longitude), El-Behaira Governorate in the 2019/2020 and 2020/2021 growing seasons. The main objectives of this study were to determine the effect of four nitrogen fertilizer levels (Control, 60, 80 and 100 kg.fed^-1), three bio-fertilizers treatments (Control, Cerealine® and T.S®) and four Molasses levels (Control, 20, 40 and 60 kg.fed^-1) on yield and quality of sugar beet plants. A split-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, as well as molasses treatments occupied the sub-sub plots. The results indicated that increasing nitrogen fertilizer rates significantly improved yield and yield components as well as the quality of sugar beet plants.  The highest rates of nitrogen (100 and 80 kg N fed^-1), bio-fertilizers treatments, (T.S^®) and molasses (60 and  40 kg N fed^-1) produced the highest yield characters (root yield (ton fed^-1), top yield (ton fed^-1) and sugar yield (ton fed^-1)) and juice quality characters (total soluble solid percentage (TSS %) and Sucrose %) throughout the 1^st and 2^nd seasons, respectively, without significant differences. The interaction between nitrogen rates, molasses and bio-fertilizers (80 kg N fed^-1+ 40 kg Molasses fed^-1+ T.S) gave the highest values for most all studied characters. So, bio-fertilizer treatments proved a major role in crop production optimization and are expected to reduce the pollution of the agricultural environment.