Plant diseases are mainly caused by plant pathogens, some of which have the ability to modify the plants genetically. Agrobacterium rhizogenes is such bacteria which has the ability to genetically modify the plant. Paederia foetida L. is one of the important medicinal plants of tremendous medicinal importance. This study aims to extract essential secondary metabolites from the roots of Paederia foetida for the design and manufacture of drugs without the plant being uprooted. Thus helps in the conservation of the plant species. We used the previously established plant tissue culture protocol as a pre-requisite for the development of hairy roots in Paederia foetida L. in this paper. Via the research aid of two Agrobacterium rhizogenes, MTCC 2364 and MTCC 532 wild type strains. In vivo and in vitro leaves and internodes were used as explants and to infection techniques viz, injecting and dipping for 15, 30 & 45 minutes were implemented. Highest (53.3 %) result was recorded in in vivo internodal explant with 30 min dipping but 15 min and 45 mins dipping showed less response (20.0%). Hairy roots induction in medicinal plants can provide the supplement to the pharmaceutical companies without disturbing or uprooting the plants, which will lead to the conservation of medicinal plants.