The invention relates to a tissue culture and propagation method of Yunnan orthosiphon aristatus and application of the tissue culture and propagation method. By utilizing the tissue culture and rapidpropagation method, wild orthosiphon aristatus leaves in Xishuangbanna of Yunnan are induced to generate a callus, and a lot of sterile tissue culture seedlings are generated after culturing of a differential culture medium, culturing of a multiplication culture medium and culturing of a rooting culture medium; leaves of the sterile tissue culture seedlings are continually subjected to callus induction, multiplication and differentiation and rooting to obtain a lot of sterile tissue culture seedlings; after open seedling exercising is carried out, the seedlings are transplanted into substratesoil containing vermiculite and soil with the ratio of 2 to 1; after the seedlings grow for 15 to 20 days, water is sprayed each day and the seedlings are managed at fixed period; after the re-growing vigor is good, the seedlings are transplanted into a large field, and the survival rate can reach 95 percent; the orthosiphon aristatus is cultured and propagated in a large scale, and the plantingarea of the orthosiphon aristatus can be expanded for hundreds of times; the requirements of markets are met, and the rapid development of an orthosiphon aristatus industry is propelled.