Arrange the following steps of somatic hybridisation in a correct sequence.

A. Digestion of cell walls.
B. Isolation of naked protoplasts.
C. Fusion of protoplasts to get hybrid protoplast.
D. Isolation of single cells from two different varieties of plants.
E. Growing of hybrid protoplast to form a new plant.

Choose the correct answer from the options given below :

Somatic hybridisation is a technique used to combine the genetic material of two different plant varieties at the cellular level. The correct chronological order of its procedural steps is:

1. D. Isolation of single cells from two different varieties of plants.
First, cells are taken from each parent plant. These cells must be healthy and viable, providing the source material for the subsequent hybridisation.

2. A. Digestion of cell walls.
To manipulate the cells directly, the rigid cellulose cell walls are enzymatically removed, typically using cellulase and pectinase. This creates wall‑less cells (protoplasts) that can be fused.

3. B. Isolation of naked protoplasts.
After digestion, the liberated protoplasts are gently separated from the enzyme solution, usually by centrifugation and washing, yielding clean, naked protoplasts ready for fusion.

4. C. Fusion of protoplasts to get hybrid protoplast.
Protoplasts from the two varieties are brought together and induced to merge, often by applying an electrical pulse (electrofusion) or polyethylene glycol (PEG). The resulting hybrid protoplast contains the combined nuclear genomes of both parents.

5. E. Growing of hybrid protoplast to form a new plant.
Finally, the hybrid protoplast is cultured in a suitable medium to regenerate a cell wall, develop into a callus, and subsequently differentiate into a whole plant that carries traits from both original varieties.

This sequence D → A → B → C → E reflects the logical progression from obtaining source cells to producing a fully formed hybrid plant.