The invention discloses a breeding method of a polyclonal antibody cold-resistant two-line sterile line, belonging to the technical field of breeding for disease resistance. According to the breeding method, by using a two-line sterile line as a recurrent parent, and using a tryporyza incertulas-resistant variety, a rice blast-resistant variety, a stunt-resistant variety and a cold-resistant variety as antigen parents, through a hybrid polymerization process, antigen genes of four rice diseases, insect and cold damages are polymerized in the same rice material, and then subjected to multi-generation backcross and screening together with the recurrent parent of the two-line sterile line, so that the insect and cold-resistant two-line sterile line, with economical characters, sterility period translative temperature and sterility highly consistent with those of the recurrent parent of the two-line sterile line, is bred. The two-line sterile line is used for preparing two-line hybrid rice, and thus the level of resisting insects and low-temperature coldness of the hybrid rice can be improved.