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Canna (Canna indica L.) is an ornamental landscape plant used specially for the garden borders and beds. It grows in tropical and subtropical countries including India. Canna is a less explored crop, mainly because it is a slow growing monocot with extremely hard seed coat and difficult to establish in vitro, as bacterial contamination is carried through the soil-grown rhizome. Many cultivars (ca. 150) of canna are being maintained in the garden germplasm of National Botanical Research Institute. To obtain 100% in vitro seed germination, chipping off of seeds with a sterilized nail clipper and soaking for 24–48 h or until radical emergence was a prerequisite. To obtain a foolproof tissue culture protocol of canna, in the present study, shoot multiplication was obtained through rhizome axillary buds. Among semisolid, liquid submerged and liquid media with glass beads, the highest multiplication of shoots (10) was obtained in liquid media with glass beads in ‘Canna Flaccida’ cv. within 6 weeks of culture incubation. During a comparative analysis of shoot regeneration among ten most attractive selected cultivars of canna, two did not respond, whereas a significant difference was obtained among eight cultivars. The regenerated shoots were rooted, acclimatized, and transferred to the pots, where they grew normally.