Mechanical Method, Plasmolyticum, Plasmolysed Cells, Protoplasts Shrink, Cell Wall, Deplasmolysed, Onion Bulbs, Scales, Radish Roots

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Protoplast Culture Regeneration and Somatic Hybridization Protoplast Culture Regeneration and Somatic Hybridization - Introduction Isolation of Proplasts Mechanical Method Enzymatic Method Direct One Step Method Sequential Two Step Method Purification of Isolated Protoplasts Sedimentation and Washing Flotation Viability and Plating Density of Protoplasts Testing Protoplast Viability Minimum Plating Density MPD and Culturing Individual Protaplasts Protaplast Culture and Regeneration of Plants Isolation of Subprotoplasts Protoplast Fusion and Somatic Hybridization Techniques of Protoplast Fusion

Home >> Plant Biotechnology and Genomics >> Protoplast Culture Regeneration and Somatic Hybridization >>Mechanical Method

Mechanical method

In mechanical method, cells are kept in a suitable plasmolyticum (in plasmolysed cells, protoplasts shrink away from cell wall) and cut with a fine knife, so that protoplasts are released from cells cut through the cell wall, and the tissue is again deplasmolysed.

Principle of Mechanical Method Isolation when tissue is cut along the dotted line, with a razor blade, some protoplasts will be released

A. Tissue is cut along the dotted line	B. Release of Protoplasts from Damaged cells

This method is suitable for isolation of protoplasts from vacuolated cells (e.g. onion bulbs, scales, radish roots). However, this method is not suitable for isolating protoplasts from meristematic and less vacuolated cells, where it gives poor yield of protoplasts.

The mechanical method, was, though, used as early as 1892, is now only rarely used for isolation of protoplasts.

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