Desmodium triquetrum DC possess immense pharmaceutical value. Preliminary phytochemical examinations were carried out before in vitro micropropagation. A rigorous attempt has been made for development of micropropagation procedure for this species, involving five steps, namely: in vitro seed germination, shoot multiplication, callus induction, rooting and hardening. In this study, direct shoot development and callus formation were achieved from different nodes and leaf disc of in vitro seedling of Desmodium triquetrum. Murashige and Skoog’s (MS) medium supplemented with different concentrations of cytokinin and in combination with auxin were used for shoot multiplication, callus induction and root formation. Polyphenolic exudation from the cultured explant was controlled by the incorporation of citric acid to the medium. The rooted plantlets were transplanted to a plastic bags containing soil mixture and grown in a green house conditions for adaptation to natural environment.