Chapter 10 Isolation and Culture of Protoplasts

Abstract

This chapter provides an overview of the isolation and culture of protoplasts. The use of protoplasts in somatic hybridization and genetic manipulation experiments is dependent on the ability to regenerate plants from isolated protoplasts, particularly in the species of agronomic and economic importance. Most of the attempts to culture protoplasts have also been limited to mesophyll protoplasts. In many cases, particularly in legumes, cell masses and callus tissues can be obtained from protoplasts. The physiological and morphogenetic status of the source material, whether plants or cultured cells, is of utmost value in determining not only the fate of the cultured protoplasts, but also the regeneration of plants from the resulting cell masses. Embryogenic tissue cultures obtained from immature embryos or other sources are the most significant source of protoplasts for plant regeneration. This proves to be the key factor in the successful regeneration of plantlets, through somatic embryogenesis, in protoplast-derived cell masses of the cereal, P. americanum .