Protoplast regeneration and organogenesis from pea protoplasts

Protoplasts of young Pisum Sativum L. seedlings from 7 different genotypes were isolated and regenerated to the callus stage. Germinating embryos were cultivated with cotyledons removed, thus avoiding intracellular starch accumulation in donor tissue. The first lateral shoots provided a source of homogenous meristematic cells which gave rise to sustained protoplast division and resulted in callus formation within 4 weeks. Root formation occurred on hormone-free medium and shoots developed on medium containing kinetin, 2iP or zeatin in the third subculture, when subcultured in monthly intervals.