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

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% (w/v) 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.