| dc.description.abstract | Pea (Pisum sativum) is a versatile and valuable crop with a wide array of applications,
including its role as a food source, forage, and even a potential biofuel crop. Given the
increasing interest in pea research, the development of an efficient method for transient
transformation holds paramount importance. In this study, we introduce a comprehensive
protocol for the isolation and transfection of pea primary root protoplasts.
Through exploration of the protoplast isolation technique, enzyme compositions and
concentrations, we have successfully achieved the isolation of pea root protoplasts, yielding
7.169E+03 protoplasts · root -1 after 6 hours of digestion. Moreover, we have identified
conditions that sustain high protoplast viability for up to 16 hours.
Furthermore, by optimizing the polyethylene glycol (PEG) calcium-mediated transformation
process, we have achieved an average transfection efficiency of 15.48% by subjecting
protoplasts to a 30% (w/v) PEG4000 solution for 5 minutes.
In conclusion, we are confident that this protocol will find wide-ranging applications and
prove highly beneficial for future research focused on the potential of pea crops for various
purposes, enabling investigations such as gene expression and protein subcellular
localization, and ultimately advancing our knowledge and utilization of this invaluable plant
species. | |
