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The hairy roots of Crotalaria sessiliflora were induced from the tissue segments infected with Agrobacterium rhizogenes ATCC 15834. The induced hairy roots were subjected to paper electrophoresis fro the detection of opine-positive clones which were considered to have been transformed. Mannopine and agropine were presented in hairy root clones while mannopine was presented in two hairy root clones. Eight hairy root clones were selected and cultured in MS, B5 and WP media. Each of hairy root clones was showed a difference in branch pattern and growth rate. The best culture medium and culture conditions of hairy roots were in MS(3% sucrose, pH 5.7) liquid medium at 25℃, 70 rpm under dark, the growth rate in MS liquid medium was increased with 210-fold more than that of inoculated hairy roots and with 2-fold more than that in MS liquid medium. Also, the adequate condition for hairy root growth was such that concentration of KH$2PO$4 was 1.25mM and the ratio of NH4 : NO was 1 to 3 in MS medium. The presence of pyrrolizidine alkaloids, monocrotaline, in the hairy roots was detected by TLC.