Seeds obtained from mature green capsules (‘pods’) of a highly fragrant and floriculturally important epiphytic orchid, Dendrobium chrysotoxum Lindl. were aseptically grown on agar gelled KC, MS, MVW, and PM media with three different sources of carbohydrates i.e. glucose, lactose, and sucrose; high response was achieved on sucrose containing PM medium. Highest mean increased seedling length (2.95±0.19 cm) was achieved on agar gelled MS medium with 0.5 mgl-1 IAA + 1.0 mgl-1 BAP followed by next seedling length (2.93±0.14 cm) on liquid MS medium with 1.0 mgl-1 IAA + 1.0 mgl-1 BAP. The pseudobulb segments directly produced highest average number of multiple shoot buds (7.04±0.46/segment) on MS medium with 1.0 mgl-1 IAA + 2.0 mgl-1 BAP followed by next response of shoot buds (6.76±0.37 segment) on MS medium with 1.0 mgl-1 NAA + 2.0 mgl-1 BAP. Leaf segments showed proliferation and produced maximum per cent of greenish PLBs (Protocorm Like Bodies) on MS medium with 1.0 mgl-1 IAA + 2.0 mgl-1 KN followed by MS medium supplemented with 1.0 mgl-1 IBA and 2.0 mgl-1 BAP. The highest mean increased individual shoot bud length (2.73±0.21 cm) was achieved on liquid MS medium with 1.0 mgl-1 IAA + 1.0 mgl-1 BAP followed by next shoot bud length (2.65±0.17 cm) on agar gelled MS medium with 0.5 mgl-1 NAA + 1.0 mgl-1 BAP. The increase in length as well as the number of roots developed in seed originated seedlings and shoot bud derived plantlets were best on MS medium with 1.0 mgl-1 IAA followed by MS medium with 0.5 mgl-1 NAA. The highest number of shoot bud formation took place in agar gelled PM medium with 0.5 mgl-1 NAA + 1.0 mgl-1 BAP and liquid MS medium with 0.5 mgl-1 Pic + 1.0 mgl-1 BAP. This protocol is a reliable way for the in vitro germination, micropropagation, seedlings development, and multiple shoot bud production of Dendrobium chrysotoxum.