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The application discloses a preparation method of super-high thermal conductive short-cut mesophase pitch-based graphite fiber and belongs to the technical field of fiber materials. The mesophase pitch fiber is obtained through twice melt spinning treatment, then the mesophase pitch fiber is cut into fixed-length fibers and subjected to non-fusion treatment, tension is applied to the obtained non-fusion fiber during carbonization treatment, then the non-fusion fiber is cut, and finally, graphitization treatment is performed. The mesophase pitch is used as raw material, twice melt spinning is performed, the fiber with more highly ordered aromatic molecular arrangement can be obtained by not adding a filter screen during the second spinning, the aromatic macromolecule can be converted into more regular carbon layers by applying tension during the carbonization process, the orientation of graphite microcrystals is favorable, and thus the super-high thermal conductive short-cut mesophase pitch-based graphite fiber can be obtained, and the carbonization is followed by short cutting, which is helpful to maintaining the orientation structure of the graphite microcrystals. The thermal conductivity of the short-cut mesophase pitch-based graphite fiber can be up to 1358 W / (m*K).