The invention provides a method for preserving the callus of rubber plant anther with vitrification and super low temperature, comprising the following steps: the anther callus is inoculated to the pre-culture medium, dark culture is carried out for 2 to 3 days with the culture temperature of 25 to 28 DEG C; the anther callus is processed by 60 percent of PVS2 for 10 to 30 minutes at room temperature; after dehydration process, the anther callus is processed by the PVS2 for 20 to 40 minutes at the temperature of 0 DEG C, the stock solution of the PVS2 is removed, then new PVS2 is added, and the anther callus is stocked in liquid nitrogen; and after the anther callus is resurgent, successive transfer culture is carried out, the regeneration plant can be obtained after further embryoid is induced and the mature embryoid is induced to carry out differentiation. The preservation method by liquid nitrogen with vitrification and super low temperature related by the invention has the advantages of simple equipment, simple experiment procedure and good effect and repetitiveness, etc., can preserve the callus of rubber plant anther for long time, obtains the regeneration plant by culturing after unfrozen and creates conditions for the genetic transformation of the rubber plant.