Conventional agricultural practices, which rely heavily on polluting agrochemicals, are pushing us towards an unsustainable future. Biofertilizers are multifaceted and span the environmental, agricultural, and economic dimensions. Therefore, this study was conducted to investigate the impact of plant growth-promoting bacteria Azotobacter chrocooccum and Pseudomonas sp. and their interaction on the growth and productivity of cotton (Gossypium barbadense L. vr. Giza CV 97) under different levels of nitrogen fertilization (50, 75, and 100% nitrogen recommended dose). Nitrogen deficiency resulted in lower cotton growth, chlorophyll, and stem anatomical parameters as well as yield or yield components compared to optimal nitrogen fertilization. Application of A. chrocooccum and Pseudomonas sp. and their interaction mitigated harmful nitrogen deficiency stress. Cotton fiber quality measurements (fiber length, micronaire, and fiber strength) were insignificantly affected by bacterial biofertilizers and their interactions under all levels of nitrogen fertilization. A. chrocooccum and Pseudomonas sp. interaction was the best treatment at all levels of nitrogen fertilization, producing thicker, wider vascular conductive, cortical tissue, and xylem vessel diameters. This treatment also improved cotton growth and gave the highest seed yield, lint percentage, and boll weight.