In the present book, tissue culture studies on two medicinally important plant species- Bacopa monnieri and Tylophora indica is presented which was carried out as a work for M.Sc. in 2010. Tissue culture methods are now routinely adopted for propagating medicinal plants as they are being depleted from wild. In vitro propagation includes five different stages and stage-III is rooting of microshoots in static or liquid media. It is an important stage as after that the in vitro plants are transferred to the field. Rooting can also be induced by using natural planting substrates which is advantageous, as it reduces the time and cost of in vitro plants. In the present work the potency of natural planting substrates for induction of roots in microshoots of Bacopa monnieri and Tylophora indica was studied. The substrates could induce roots in the shoots and were suitable in initiating further growth in them. Hence these substrates can be an alternative to the rooting medium in micropropagation. This book can help the researchers to adopt the use of planting substrates for rooting in other species.