Last 3 Steps of Protoplast Fusion | Callus Formation & Plant Regeneration.

[00:00:00]Steps in protoplast fusion. There are six steps in protoplast fusion. One, isolation of protoplast. Two, fusion of protoplasts. Three, selection of hybrid cells. Four, cell wall regeneration.

[00:00:16]Five, callus formation. Six, plant regeneration.

[00:00:21]Next, we are going to see about cell wall regeneration. After selection, the hybrid protoplast starts forming a new cell wall. Cellulose and pectin materials are deposited around the plasma membrane. The new cell wall gradually develops and becomes complete.

[00:00:39]Cell wall regeneration helps the cell regain its shape and stability. A regenerated cell wall is essential for cell division and callus formation.

[00:00:50]Next, callus formation. After cell wall regeneration, the hybrid cells start dividing continuously. Repeated cell divisions produce a mass of undifferentiated cells called callus.

[00:01:03]Callus formation takes place on a nutrient culture medium.

[00:01:08]Plant growth regulators like auxins and cytokinins help in callus development.

[00:01:13]The callus can later regenerate into roots, shoots, and a complete plant. The last step is plant regeneration. The callus is transferred to a suitable regeneration medium. Shoots are first developed from the callus under controlled conditions. Roots are then formed from the regenerated shoots. A complete plantlet with shoots and roots is produced. The plantlet is transferred to soil, where it grows into a mature plant.