The invention discloses a preparation method for an aseptic explant of a virus-free seedcane of sugarcane. The preparation method comprises the following steps: cutting a plant which grows robustly and does not have diseases and insect pests into single-bud sections; planting the single-bud sections in an aseptic provenance garden after the single-bud sections are soaked for 2 hours in a constant temperature field of 52 DEG C; after 5 to 7 months, taking the tips of pathogen-free plant sugarcane after more than or equal to 3 consecutive sunny days, and cutting the tips of the stems to serve as an aseptic explant by adopting an aseptic operation. The biggest difference between the preparation method and the conventional external building method is that the mercury chloride sterilization step is omitted. The preparation method is ecological, environmentally friendly, simple and feasible, and provides a good aseptic explant material taking method for the scale production of the virus-free seedcanes of the sugarcanes.