The invention discloses a method for cultivating aphelenchoides besseyi by using carrot callus and belongs to the technical field of plant nematodes cultivation and preservation. The method includes specific steps of washing fresh carrots clean, soaking in alcohol, firing unit surfaces of the carrots become dark red, peeling the surfaces, cutting into slices, placing in a sterilization petri dish containing water agar, sealing, placing in an incubator, and cultivating until the carrot callus is grown out; disinfecting the aphelenchoides besseyi by using streptomycin sulfate solution, centrifuging, precipitating, evenly mixing by using sterile water, centrifuging continuously, finally precipitating, diluting by using the sterile water, and dropping on the carrot callus in dispersing mode; and placing the carrot callus with inoculation of the aphelenchoides besseyi in a black incubator, and cultivating at the temperature of 25 DEG C-30 DEG C or preserving at the temperature of 20 DEG C-25 DEG C. The method for cultivating the aphelenchoides besseyi by using the carrot callus can cultivate and reproduce the aphelenchoides besseyi simply and largely and can also maintain parasitic and pathogenic force of the aphelenchoides besseyi to the plant.