Plant Regeneration from Protoplasts Isolated from Callus of Taro (Colocasia esculenta Schott)
1995 Volume 63 Issue 4 Pages 773-778
Details
1995 Volume 63 Issue 4 Pages 773-778
A system of protoplast isolation, callus formation, and plant regeneration in taro (Colocasia esculenta Schott cv. Eguimo) was established.
1. Friable calli were induced by culturing etiolated stem segments of taro on Murashige and Skoog (MS) medium supplemented with 30g•liter-1 sucrose, 2 mg•liter-1 2, 4-D and 2 mg•liter-1 2ip. Calli were maintained by subculturing on a fresh MS medium.
2. Protoplasts were easily isolated from suspension cells derived from the friable calli. The component of the enzyme solution for the isolation consisted of 1g•liter-1 Pectolyase Y-23, 5g•liter-1 Cellulase Onozuka RS, 5 mM MES, 5 mM CaCl2•2H2O, and 0.5M mannitol.
3. Isolated protoplasts were cultured in the liquid media consisting of half strength MS inorganic salts, Kao and Michayluk's (1975) organic substances, various levels of NAA and BA, 0.1M glucose and 0.3M mannitol. Numerous colonies were formed in the medium containing 2mg•liter-1 BA.
4. Shoot regeneration from protoplast-derived calli occurred on solid MS medium with 0.2mg•liter-1 NAA and 2 mg•liter-1 BA. The shoots initiated roots after being transferred to a MS medium without phytohormones.
The protoplast culture system established in this study might be useful for cell fusion and electroporation of genes as an approach to breeding of taro.
Chemical names used: 2-N-MorPholino ethanesulfonic acid (MES) ; 1-naphthanlenacetic acid (NAA) ; 2, 4-dichlorophenoxy acetic acid (2, 4-D) ; N-phenylmethyl-1H-purin-6-amine (BA) ; N-3-methyl-2-butenyl-1H-purin-6-amine (2ip).
