Desarrollo de un protocolo para extracción y fusión de protoplastos de solanum tuberosum (papa) y solanum lycopersicum (tomate)

Abstract

Protoplast fusion is a crucial biotechnological technique that enables the combination of characteristics from different cells, advancing research and the development of new plant varieties. This study evaluates the impact of varying concentrations of polyethylene glycol (PEG) and pH levels on protoplast fusion efficiency. Five treatments were applied: 40% PEG at pH 8, 40% PEG at pH 9, 50% PEG at pH 8, 50% PEG at pH 9, and a control with water, each with three repetitions. Protoplast fusion and non-fusion/dead counts were conducted using a Neubauer chamber, allowing clear identification of fused cells due to morphological differences between the species involved. Results indicated that both PEG concentration and pH significantly impact protoplast fusion. Notably, the 50% PEG at pH 9 treatment was the most effective, yielding the highest number of fused protoplasts. A 2x2+1 factorial analysis confirmed the significant interaction between PEG concentration and pH. The control treatment, without PEG, demonstrated the lowest efficiency, highlighting the critical role of PEG in promoting cell fusion. These findings provide a solid foundation for optimizing protoplast fusion processes in biotechnological applications, with a recommendation to use 50% PEG and pH 9 to maximize fusion efficiency.