Development of an Efficient Micropropagation Procedure for an Ornamental Aquatic Plant, Cryptocoryne wendtii through Shoot Multiplication and Plantlet Regeneration

Abstract

An in vitro plant regeneration protocol was successfully established for Cryptocoryne wendtii , an ornamental aquatic plant by culturing shoot tip explants. The shoot tips were surface sterilized using 8 % Clorox® (5.25 % sodium hypochlorite, NaOCl) for 15 minutes followed by rinsing three times with sterile distilled water. They were again surface sterilized for another 4 % Clorox® (5.25 % sodium hypochlorite, NaOCl) for 5 minutes. Of the two cytokinins, 6-benzyl-aminopurine (BAP) and Kinetin (Kin) were evaluated as supplements to Murashige and Skoog (MS) medium [1], BAP at an optimal concentration of 3.0 mg/l was effective in inducing multiple shoots. The highest shooting percentage was 100.0 % on MS medium. Shoot multiplication of C. wendtii can be induced from shoot tip explants cultured on MS medium supplemented with 3.0 mg/l BAP and significantly different from other treatments, with the highest number of 16.20 shoots per explant and number of leaves at 72.40 leaves per explant. Shoots produced an average of 36 roots per plantlet and root length at 26.02 mm when cultured on MS medium supplemented with 1.0 mg/l α-Naphthaleneacetic acid (NAA). Plants were successfully transferred into plastic pots containing a mixture of organic soil and sand (1:1) overlaid with tap water with 100 % survival rate under greenhouse conditions. The in vitro regenerated plantlets were hardened and acclimatized successfully. The current protocol is the first successful report of C. wendtii micropropagation