Isolation of Protoplasts

Abstract

Although protoplasts have been isolated from a range of somatic tissues and successfully cultured, the success in isolating the protoplasts from reproductive tissues has been rather limited. This is basically due to technical difficulties. In the pollen, for example, the exine, which is made up of sporopollenin, is resistant to enzymatic degradation; in the ovule the embryo sac is deeply embedded under a large number of cell layers and is not therefore readily accessible for release due to action of cellulytic enzymes. In recent years, however, some successful attempts have been made to isolate protoplasts from microspores, pollen grains, and embryo sacs. The techniques need to be refined, but yet they have great potential in basic studies on the biology of fertilization and in applications of plant breeding and biotechnology (see Zhou and Yang 1986; Knox and Singh 1987).