The invention belongs to the field of agricultural high technologies and specifically relates to a hemsleya amabilis tissue culture and lateral propagation method. The method comprises the following steps of initial medium preparation, explants sterilization and treatment, inoculation and culture, subculture medium preparation and inoculation, rooting culture, seedling hardening and transplantation. The method disclosed by the invention overcomes loss of effective ingredients cultured under a specific environment of a laboratory in the prior art and achieves artificial cultivation of wild varieties. Detection shows that dihydrocucurbitacin, cucurbitacin and hemsleya amabilis saponins all can be detected from the hemsleya amabilis propagated by the method disclosed by the invention, and contents of the dihydrocucurbitacin, the cucurbitacin and the hemsleya amabilis saponins are equivalent to that of the dihydrocucurbitacin, the cucurbitacin and the hemsleya amabilis saponins in wild hemsleya amabilis. The method disclosed by the invention improves a qualified seedling percentage of hemsleya amabilis seedlings, shortens a cultivation period, keeps effectiveness of the wild hemsleya amabilis and has an application prospect in the aspects of preserving, protecting, developing and utilizing wild hemsleya amabilis resources and the like.