The invention discloses a Ce,V and Er co-doped dental microcrystalline glass, and a preparation method and application thereof. The raw materials of the microcrystalline glass consist of SiO2, Li2O, K2O, P2O5, Al2O3, ZrO2, Na2O, CeO2, V2O52 and Er2O3. A glass block is obtained through a melt casting method, and microcrystalline glass easy for machining is obtained after crystallization treatment,and then secondary crystallization is carried out to obtain the finished product of the microcrystalline glass which takes Li2Si2O5 as a main crystalline phase. The microcrystalline glass has excellent bending strength, good semi-permeability, fluorescent property, uniform and full coloring and excellent stability. The color and the aesthetic effect of the microcrystalline glass reach the color and the aesthetic effect of A2 in a VITA-Classical colourimetric card and are consistent with the fluorescent characteristic and the semi-permeability of natural teeth, and the microcrystalline glass can be applied to dental whole-porcelain repair materials.