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Protoplast

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Protoplast, from the ancient Greek πρωτον (first) + πλασσειν (to mould), initially referred to the first organized body of a species. This meaning is similar to the non-biological definition, the first from which all subsequent forms are derived.

Subsequently it has several biological definitions:

Enzymes for the preparation of protoplasts

Cell wall are made from a variety of polysaccharides, protoplasts can be made by degrading cell walls with a mixture of the appropriate polysaccharide degrading enzymes.
Type of cell Enzyme
Plant cells Cellulase, pectinase, xylanase
Gram-positive bacteria Lysozyme (+EDTA)
Fungal cells Chitinase
Following the digestion of the cell wall the protoplast is very sensitive to osmotic stress, and must be stored in an isotonic solution to prevent rupture.

Uses for protoplasts

Protoplast can be used to study membrane biology, including the uptake of macromolecules and viruses.

Protoplasts are widely used for DNA transformation (for making genetically modified organisms), or for studying temporal gene expression. Protoplasts may be regenerated into whole plants using Micropropagation, a modern technique employing plant tissue culture methods.

Protoplasts may also be used for plant breeding, using a technique called protoplast fusion. Protoplasts from different species are forced to fuse by using an electric field or a solution of polyethylene glycol. This technique may be used to generate hybrids in tissue culture.

 


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