The invention discloses an assembly adjustment fluorescence enhancement aggregation-induced emission material, a micro-nano sphere, a preparation method and application. The molecular formula of the luminescent material is formula I. The material disclosed by the invention is stable in structure, has aggregation-induced emission and mechanochromic properties, and is good in luminescence property, the powder state fluorescence quantum yield can reach 45.5%, and the powder fluorescence quantum yield after grinding can reach 65.6%; by adopting an aggregation method in an evaporation solvent and a poor solvent, the luminescent material can be prepared into micro-nano spheres with different sizes through self-assembly, and the fluorescence quantum yield of the nano-scale spherical assembly can reach 85.3%; the synthesis and preparation method of the aggregation-induced emission material is simple to operate, mild in reaction condition and high in yield; the aggregation-induced emission material and the assembled micro-nano spheres are insoluble in water and most of organic solvents, have high fluorescence efficiency and mechanochromic performance, and can be used in the fields of counterfeiting prevention, real-time monitoring of pressure change, biological fluorescent probes, biological imaging and the like.