Protoplast isolation, culture and plant regeneration in Butterflypea [Clitoria ternatea (Linn.)]

Abstract

This study describes successful plant regeneration using protoplasts isolated from mesophyll cells of Clltoria ternatea (Linn.). Maximum protoplast yield (7.34 x 106 g/fresh wt of leaf tissue), and viability (86.3%) was recorded with an enzyme mixture containing 1.0% (wlv) Cellulase, 1.5% (w/v) Macerozyme and 0.3% (w/v) Pectolyase. An optimum density of 2.5 x 103 protoplasts/ml was required for the highest frequency (71.25 ± 6.85%) of division. Dividing protoplasts produced microcalli that proliferated on MS medium containing 9.29 pM Kinetin and produced shoot buds when MS medium incorporated with 100 mg 1-1 adenine sulphate. Elongated shoots were rooted on half strength MS basal medium without growth regulators. The well rooted shoots were transferred and acclimatized.