Rice blast is one of the most devastating diseases in rice production. Although the commercially-released resistant cultivars often become susceptible in 3-5 years due to the rapid evolution of M. oryzae isolates, breeding for resistant variety is still the most economical and effective approach to control blast disease. In this study, the donor line R6, which harbors rice blast resistance gene Pi25, was crossed with the super early-season rice Zhongzao39 (the receptor and recurrent parent), followed by generations of backcrossing, self-pollination and molecular marker-assisted selection. The offspring lines were tested for rice blast resistance by spray inoculation in the greenhouse, injection inoculation in the field and natural infection in the disease nursery. Remarkably, four lines (i.e. FY82, FY90, FY125 and FY137) carrying Pi25 showed the elevation of resistance on leaf blast and neck blast. In addition, investigation of the genealogical composition and other resistance loci provided a preliminary dataset in order to decipher the genetic basis of rice blast resistance.