Two protoplasts can be made to fuse through the apllication of

Options

A. Electrofusion
B. Polyethylene glycol
C. Sodium nitrate
D. All the above

Answer

To solve the question, "Two protoplasts can be made to fuse through the application of," we will analyze the options provided and understand the methods of protoplast fusion.

### Step-by-Step Solution:

1. **Understanding Protoplasts**: Protoplasts are plant cells that have had their cell walls removed. This allows for the fusion of two protoplasts to create hybrid cells, which can be useful in plant breeding and genetic engineering.

2. **Identifying Fusion Techniques**: There are several techniques that can be used to induce the fusion of protoplasts:
- **Electrofusion**: This method involves applying an electric field to the protoplasts, which causes them to fuse together. The electric field destabilizes the membranes of the protoplasts, allowing them to merge.
- **Polyethylene Glycol (PEG)**: This is a chemical method known as chemofusion. PEG facilitates the fusion of protoplasts by promoting the merging of their membranes, especially when calcium ions are present.
- **Sodium Nitrate**: This method involves using sodium nitrate in a sucrose solution to suspend the protoplasts. The sodium nitrate helps in the fusion process by creating an environment conducive to merging.

3. **Evaluating the Options**:
- **Option A: Electrofusion** - Correct, as it is a recognized method for protoplast fusion.
- **Option B: Polyethylene glycol** - Correct, as it is also a method used for protoplast fusion.
- **Option C: Sodium nitrate** - Correct, as it can facilitate protoplast fusion.
- **Option D: All of the above** - Since all the previous options are valid methods for protoplast fusion, this option is correct.

4. **Conclusion**: Based on the analysis, the correct answer is **Option D: All of the above**. All three methods can be used to induce the fusion of protoplasts.