Belonging to plant cultivation technologies, the invention provides an in vitro culturing method for photinia fraseri breeding. The method comprises initiation culture, proliferation culture, and rooting culture. The method specifically consists of the steps of: initiating culture, inoculating a terminal bud or a stem segment with one axillary bud into an initiation culture medium, which is then placed in a culture room of 23-27DEG C, taking a fluorescent lamp as the light source, controlling the illumination intensity at 1500Lx-2000Lx, and conducting illumination for 10h/d and culturing for 20-30d, with the relative humidity of the culture room maintained at 70%-80%; transferring a generated adventitious bud into a proliferation culture medium, which is placed in a culture room of 23-27DEG C, taking a fluorescent lamp as the light source, controlling the illumination intensity at 1500Lx-2000Lx, and conducting illumination for 10h/d and culturing for 20-30d, with the relative humidity of the culture room maintained at 70%-80%, thus producing more adventitious buds, and transferring the robust young seedlings into a rooting culture medium, which is placed in a culture room of 20DEG C, taking a fluorescent lamp as the light source, controlling the illumination intensity at 1500Lx and the illumination time at 12h/d, maintaining the relative humidity of the culture room at 70%-80%, and carrying out rooting culture for 20-25d so as to make the length of the root up to 3-5cm.