Abstract

Sustainable agriculture and food security are challenged by the indiscriminate use of synthetic nitrogen (N2) fertilizers, inefficient water management, and land degradation. Hydroponic cultivation uses nutrient-rich aqueous media and is a climate-resilient and resource-efficient alternative to traditional farming methods, whose dependence on synthetic N2 fertilizers reduces their long-term sustainability. Biological nitrogen fixation (BNF), which is mediated by diazotrophs that reduce atmospheric N2 to plant-available ammonium, has emerged as a sustainable alternative to synthetic N2 input in hydroponic systems. This review discusses the integration of BNF into hydroponic systems by exploring the functional diversity of diazotrophs, root–microbe interactions, and environmental constraints. It further highlights recent advances in strain improvement, microbial consortia development, nitrogenase protection, and genome editing tools, novel bioformulation strategies to enhance microbial compatibility with hydroponic nutrient regimes, and omics-based tools for the real-time assessment of N2 fixation and microbial functionality. Key challenges, such as microbial leaching, nitrate-induced inhibition of nitrogenase activity, and the absence of standardized biostimulant protocols, are discussed. Case studies on staple crops have demonstrated enhanced NUE and yield productivity following diazotroph applications. This review concludes with future perspectives on synthetic biology, regulatory policies, and omics-based tools for the real-time assessment of N2 fixation and microbial functionality. © 2025 by the authors.