The invention discloses a breeding method of breeding boars with high reproductive performance. The method mainly comprises the steps as follows: a seminal fluid of breeding boars with certain breeding production records (20 births or above) is acquired, sperm and seminal plasma are separated with a percoll density gradient centrifugation method, and sperm and seminal plasma protein samples are prepared respectively; quantitative protein detection is carried out on the sperm and seminal plasma protein samples with a high-throughput iTRAQ technology, and seminal fluid differential protein related to the reproductive performance of boars is screened; the correlation between the obtained seminal fluid differential protein and the reproductive performance of breeding boars is verified again, and a batch of spermatine markers reflecting the reproduction performance of the breeding boars are determined through external fertilization tests, protein expression location tests and the like; thereproductive performance of the boars and the fertilization ability of the seminal fluid are assessed by use of the relative content of these spermatine markers, so that the breeding boars with high reproductive performance are screened out. The method is convenient to use in production, all that is required is to detect the content of the spermatine markers, and sampling injury and stress of boars can be avoided.