ABSTRACT
Plant protoplasts ('naked' cells) provides a unique single cell system to underpin several aspects of modern biotechnology. Major advances in genomics, proteomics and metabolomics have stimulated renewed interest in these osmotically fragile wall-less cells. Reliable procedures are available to isolate and culture protoplasts from a range of plants, including both mono- cotyledonous and dicotyledonous crops. Isolation of protoplast is readily achieved by treating cells tissues with suitable mixture of cell wall degrading enzymes. The enzyme digested plant cells, besides protoplasts contain undigested cells, broken protoplasts and undigested tissues. The cell clumps and undigested tissues can be removed by filtration. This is followed by centrifugation and washings of the protoplasts. The process of cell wall formation in cultured protoplasts starts within a few hours after isolation that may take two to several days under suitable conditions. Plant regeneration can be done from the callus obtained either from protoplasts or from the culture of plant organs.
