Controlling environmental parameters, including nitrogen and phosphorus use efficiency, is a crucial aspect of maintaining effective aquaponic systems. In this study, we evaluated the impact of start-ups under high (20 kg/m3) and low (5 kg/m3) stocking density conditions on water quality, productivity, nitrogen use efficiency (NUE), phosphorus use efficiency (PUE), microbial community compositions, and functional genes in the aquaponics system. Six experimental units (three replications per treatment) were categorized into two groups. The experimental process was divided into two phases: Phase I (Days 1–40) and Phase II (Days 41–145). During Phase I, the two system groups were initiated under high and low stocking densities. Phase II constituted the fish and vegetable production phase, with all aquaponics systems standardized at a stocking density of 10 kg/m3 and a planting density of 12 plants/m2. The results indicated superior water quality in the low aquaponics systems than that in the high aquaponics systems during Phase I, whereas the opposite was observed during Phase II. The final weight, specific growth rate, and fish weight gain of jade perch (Scortum barcoo), as well as the plant weight gain of tomato plants, were significantly higher in the high aquaponics system than those in the low aquaponics system. The NUE and PUE of the high aquaponics system were 49.55 ± 1.02% and 60.12 ± 1.28%, respectively, surpassing those of the low aquaponics system (43.41 ± 2.16% and 49.06 ± 1.59%). The microbial diversity in the moving bed biofilm reactor and roots of the high aquaponics system was lower than in the low aquaponics system. However, the relative abundances of nitrifying and denitrifying bacteria were higher than those in the low aquaponics system. The number of copies of functional genes for nitrification and denitrification, including amoA, nxrB, nirS, nirK, and nosZ, was higher in the high aquaponics systems. Overall, the results indicated that high aquaponics systems initiated with high stocking density yield more products, improved water quality, and higher NUE and PUE by altering the composition of microbial communities. This study presents a method for enhancing the functional microbial community of a system by optimizing conditions during the initiation period.