The increasing biogeochemical imbalance of nitrogen (N) heightens the importance of studying rhizosphere bacteria, which aid crop nutrient uptake, and their responses to N deficiency. The aim of the study was to assess variety-specific responses of the tomatoes and their associated rhizobacteriome to low N availability. Three determinant varieties of Solanum lycopersicum L. were cultivated in pot-scale experiment during 10 weeks on low-fertility substrate (sod-podzolic soil:peat:clay:sand, 1:1:1:2, v/v), half of which were supplemented with ammonium nitrate (60 mg N kg⁻¹ in total). A comprehensive methodology was employed, including 16S rRNA metagenomic Nanopore sequencing, quantitative assessment of N-cycling bacteria, and analysis of plant growth, photosynthetic pigments, total N in biomass, and fine root architecture. Results demonstrated that N deficiency significantly reduced plant biomass and photosynthetic pigments. The rhizosphere contained a diverse community of N-transforming bacteria (38 identified genera), whose composition and relative abundance were strongly influenced by both tomato variety and N fertilization. Nitrogen application increased the abundance of N-fixers and altered alpha-diversity in a variety-dependent manner. Significant correlations were found between the abundance of key bacterial genera (e.g., Stenotrophomonas, Rhizobium) and N parameters in plants and substrates. The study concludes that the response of the tomato rhizobacteriome to N availability is variety-specific, which is important for the development of microbiome management strategies for enhancing N use efficiency.