The invention discloses rice herbicide resistant protein and application thereof in plant bleeding. The ALS (acetolactate synthase) protein provided by the invention realizes that a plant has the herbicide resistance, and is derived from rice varieties Huangsizhan and Huanghuazhan, and carries Trp548, Ala96 or Ser627mutation. Furthermore, the invention also provides corresponding nucleic acid, expression cassettes, carriers, cells, plants, and method for obtaining the plants with herbicide resistance and application of the plants, as well as a method for identifying the plants in the invention.