Overview of protoplast fusion workflows Which sequence best describes what the protoplast fusion technique generally involves in plant biotechnology?

Introduction / Context:
Protoplast fusion enables somatic hybridization, combining genomes from sexually incompatible species or genotypes. The classic workflow proceeds from wall removal to fusion and then to culture-based regeneration.

Given Data / Assumptions:

• Cell walls are enzymatically removed to yield protoplasts.
• Fusion is induced chemically (PEG) or electrically (electrofusion).
• Fused cells are regenerated into plants via callus/organogenesis.

Concept / Approach:
The technique does not end at fusion; viable hybrids require culture steps to reform walls, divide, form callus, and regenerate shoots/roots. Selection markers and cytological checks verify hybridity.

Step-by-Step Solution:

Verification / Alternative check:
Hybridity confirmed by chromosome counts, molecular markers, or organelle-specific markers (cybrids).

Why Other Options Are Wrong:

• Isolation alone is incomplete.
• Isolation+fusion without culturing will not yield plants.
• Conventional crossing is sexual, not somatic fusion.
• Electroporation of intact tissues is a different method (gene transfer), not fusion.

Common Pitfalls:
Failing to optimize osmotic conditions; inadequate selection allows unfused cells to dominate.

Final Answer:
Isolation, fusion, and culturing (regeneration)