5. All of the following are reasons for purifying protoplasts before culture except All of the...

5. All of the following are reasons for purifying protoplasts before culture except

All of the above are valid reasons
Remove non-totipotent cells
Remove cellular debris
Remove non-protoplasted cells
Remove all traces of protoplasting enzymes

Expert Solution

Q.5. Answer- Remove non-totipotent cells

Explanation-

Remove non-totipotent cells- not all plant cells are totipotent. So that for protoplast culture the non- totipotent cells are removed from sample. Totipotent cells are the cells that can regenerate in cultural conditions. Before protoplast culture non-totipotent cells can not be identified and removed. When protoplast is inoculated in media containing all components required for growth and even though protoplast cannot be divide and regenerate then they are non-totipotent cells. And these cells are not cultivable in tissue culture. So that these non-totipotent cells can not be removed before culture.

Reasons for purifying protoplast before culture are-

Rrmove cellular debris- During protoplast isolation mechanical and enzymatic methods are use to isolate protoplast. But some cells gets ruptured during process or some cells release broken dead protoplast. So that before culture these cellular debries are removed by filtering sample through nylon mesh.

Remove non-protoplasted cells - non-protoplasted cells are the cells whose cell wall is not broken to release protoplast. So that before protoplast culture these non-protoplasted cells are removed from protoplasted cells.

Remove all trees of protoplasting enzymes- during protoplast isolation enzymes are used to degrade cell wall. After degradation of cell wall protoplast is isolated by centrifugation. By centrifugation, protoplast settles at the bottom of tube and supernatant contains enzymes is removed by pipette.

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