Acorus calamus is an important medicinal plant which has been used in Indian traditional medicine since time immemorial. Various bioactive molecules, viz., acorin, ${alpha}$- and ${eta}$-asarone, asaryldehyde, caryophylene, isoasarone, methylisoeugenol, and safrol have been isolated from this plant. However, the use of this plant for medicinal purpose has been recently banned due to the high toxic property of ${eta}$-asarone. The triploid Acorus calamus is reported to be low in ${eta}$-asarone content and thus found to be the ideal raw material for medicinal use. The present investigation represents our finding for successful in vitro clonal propagation of the elite triploid accessions of Acorus calamus for mass propagation. In the dual-phase culture system consisting of agar-solidified Murashige and Skoog medium overlaid by liquid fraction of the same medium, maximum multiple shoot induction was favored by supplementation of ${alpha}$-naphthaleneacetic acid (0.5 mg $L^{-1}$) and 6-benzylaminopurine (2.0 mg $L^{-1}$). In vitro rooting of the micros hoots was maximum in the medium supplemented with indolebutyric acid at 2.0 mg $L^{-1}$. The well-rooted microshoots could be successfully hardened and transplanted in the field. This result can be reproduced and is a viable protocol for successful clonal propagation of the seedless triploid Acorus calamus for conservation and sustainable development.