Advancing plant protoplasts: innovative techniques and future prospects

Abstract The agricultural industry is facing increased pressure due to climate change and a growing population. Crops must be fortified with genetic advantages that will allow them to survive the effects of climate change. Protoplasts, an experimental plant system with regenerative capabilities, is one of the anticipated new plant breeding technologies (NPBT). These isolated dedifferentiated plant cells have the potential to undergo cell division and regenerate into a complete plantlet. They can be fused, transformed, or edited, making them ideal models for investigating plant cell cycle, cell wall biogenesis, organogenesis, and totipotency. Protoplast-based systems present unique advantages and challenges in crop improvement. In this mini-review, we will explore the steps required for isolating and culturing protoplasts, as well as the potential and applications of an experimental plant-based protoplast system. Cutting-edge methods in protoplast isolation, encompassing enzymatic, mechanical, and high-throughput approaches, have streamlined the release of protoplasts from plant cells. These advancements are instrumental in expediting genetic modification processes, enabling the harnessing of plant cells for biotechnological applications. The integration of CRISPR/Cas9 technologies amplifies the precision and speed of genetic modifications, fostering a new era of tailored cells for bioproduction and space research.