The invention belongs to the technical field of tobacco breeding and plant tissue culture and particularly relates to a method for inducing a tobacco haplobiont by adopting a one-step method. The method comprises the following steps: (1) low-temperature pretreatment of flower buds; (2) disinfection and sterilization; and (3) inoculation on an improved MS culture medium and culturing. The technological idea for inducing the tobacco haplobiont comprises the following steps: performing dedifferentiation on anthers; forming an embryoid; and directly forming an intact pollen plant. As the culture mode is relatively close to the development process of plants, the frequency of inducing the haplobiont is relatively high, and a plurality of pollen plants can be induced from one anther. Meanwhile, the induction time required for obtaining the haplobiont is relatively short and the haplobiont can be obtained within a month. The integral culture steps are further relatively simple and convenient, so that the method has relatively good popularization and application values in tobacco cross-breeding.