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Various technologies, such as light-emitting diodes (LEDs) and beneficial plant micro-organisms, have been applied to enhance plant growth and development. We aimed to develop appropriate technology by incorporating the benefits of LED light, plant growth promoting rhizobacteria (PGPR), and arbuscular mycorrhiza fungi (AMF) into sweet girl cherry tomato (Solanum lycopersicum L.) seedling production. Our results demonstrated that incorporating red (R) and blue (B) LED lights, PGPR, and AMF positively affected tomato seedling growth. The optimal lighting conditions for tomato seedling growth were LEDs at 200 µmol/m2/s with a ratio of R60:B40 and 20 h/d exposure. The optimum LED-illuminated tomato seedlings significantly upregulated photosynthesis-related genes, including psbA, psbB, fdx, atpB, and rbcL. Plants inoculated with PGPR Bradyrhizobium sp. SUTN9-2, Bacillus velezensis SD10 and B. megaterium A20 had a high health index after inoculation. Furthermore, the optimized LED-illuminated tomato seedlings inoculated with SD10 had the highest health index. In addition, the optimum LED-illuminated tomato seedlings inoculated with SD10 and AMF had the highest biomass. Our experiment demonstrated that tomato seedlings produced under optimized LED lights inoculated with SD10 and AMF increased yield by about 16% under field conditions. Therefore, these results provided the optimum conditions for a high-quality tomato seedling production system. [ABSTRACT FROM AUTHOR]