The invention discloses a method for identifying whether wheat to be tested is Pina-D1b deficient wheat and applications thereof. The method according to the invention comprises the following steps: 1) using a specific primer pair for PCR amplification by taking genome DNA of the wheat to be tested as a template, wherein the specific primer pair is the primer pair composed of nucleotide sequences, which are shown in sequence 1 and sequence 2 of sequence table; and 2) detecting PCR-amplified product, wherein the wheat to be tested with 776bp strips in the amplified product is candidate Pina-D1b deficient wheat, and the Pina-D1b deficient wheat is the wheat having nucleotides, which are shown in sequence 3 of the deficient sequence table of the genome DNA. The specific primer pair accordingto the invention can take the place of traditional SDS-PAGE and two-dimensional electrophoresis technology and the detection for Pina-D1b type is directly carried out at molecular level, so accuracy and reliability are greatly enhanced, screening efficiency and accuracy for grain hardness genotype is remarkably quickened; in addition, the method plays an important role in improving gene engineering of wheat grain hardness, simultaneously aids in further research on genetics and molecular biology of wheat grain hardness and is also significant to the cultivation of high-quality wheat.