The invention provides a preparation method and an application of an attapulgite/nano-iron composite material-microorganism coupled body. The preparation method comprises the following steps: purifying attapulgite with distilled water and sodium hexametaphosphate, modifying by hydrochloric acid, conducting a centrifugal process, washing, drying, putting into a FeSO4 solution, soaking for 24-72 hours, conducting the centrifugal process, adding ethanol into a solid after the centrifugal process under the protection of argon gas, uniformly mixing, dropwise adding KBH4 for full reaction of Fe<2+>, conducting the centrifugal process, washing and drying deposits after the centrifugal process, and producing a coupling reaction with alcaligenes eutrophus to obtain a solid matter as the attapulgite/nano-iron composite material-microorganism coupled body. According to the coupled body, the shortcomings of poor nano-iron stability, poor dispersity, high possibility of conglomeration and the like are overcome, the defect of secondary pollution on a water body due to ammonia nitrogen generated during reaction of an attapulgite/nano-iron composite material with nitrate is avoided, and microorganisms basically cannot be poisoned.