| Abstract | Somatic cell hybridization is a potentially useful technique for the introduction of genetic variability into plant species. One proposed method involves fusion of plant protoplasts (which is a cell lacking its wall—defined here on the basis of light microscope examination) to form a hybrid cell which can be induced to form a plant. Plant protoplasts produced by the use of crude, commercial lytic enzymes are not always healthy (as defined by consideration of factors such as cytoplasmic streaming, presence of cytoplasmic strands and position of the nucleus within the cell, in relation to survival time). Peroxidase, basic proteins and ribonuclease have been implicated as possible deleterious agents. The use of purified enzymes offers a more rigorously defined system for obtaining healthy protoplasts. We have used purified preparations of cellulase (EC 3.2.1.4), pectate lyase (EC 4.2.99.3) and endopolygalacturonase (EC 3.2.1.15) to prepare protoplasts from a range of plant cell suspension cultures. We have also prepared inducible streptomycete enzymes lytic towards plant cell walls. |
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