This study was conducted during 2015 & 2016 seasons, on 16 years old Thompson Seedless grapevines, grown on the experimental orchard of the Faculty of Agriculture, Assiut University to study the effect of bio-fertilization (nitrobine or EM) on vegetative growth, nutrient status, and fruiting of Thompson Seedless grapevines. The experiment was arranged in a complete randomized block design with five treatments and three replications one vine per each. The obtained results could be summarized as follow: Using the recommended dose of nitrogen (RDN) via 75 or 50% mineral plus 25 or 50% bio-form significantly increased, pruning wood weight, number of leaves/shoot and leaf area as well as leaf total chlorophyll and leaf nutrient composition compared to use RDN via mineral N fertilizer alone. No significant differences on these traits due to use RDN via 50% mineral plus either 12.5 or 37.5% of nitrobine or 37.5 or 12.5% effective microorganisms (EM) were noticed. All combined fertilization treatments significantly increased the yield and improved the cluster and berry traits compared to use RDN via mineral source only. It is evident from the forogoing results that double form 50% mineral plus either (37.5 and 12.5% nitrobine) or (12.5 or 37.5% EM) improved the vegetative growth, yield and berry quality. In addition, it minimized the production costs and reduced environmental pollution.