Callus formation from mesophyll protoplasts of Pisum sativum

Abstract

Protoplasts isolated from mesophyll of Pisum sativum L. cv. Century, and cultured in 0.2-ml droplets of B5 medium with 1.0 mg/liter (2,4-dichlorophenoxy)acetic acid (2,4-D) or 1-naphthaleneacetic acid (NAA) and 2.0 mg/liter kinetin regenerated cell walls within 2–3 days. The resulting cells began to divide and form calli after 19 days of culture. Protoplast survival depended on keeping the leaf material in the dark for at least 30 h before use.