Table 1.

Troubleshooting

Problem	Possible reason	Solution
Low protoplast yield and low transfection efficiency	Condition of suspension cells	Suspension cell condition is the most important factor for obtaining successful results. If you have problems with a transfection, prepare new suspension cells from your stock or rice seeds
	Insufficient cellulase digestion	Optimize the period of cellulase treatment or prepare fresh cellulase solution
	Problem with transfection reagents	Prepare new transfection reagents. PEG solution can not be stored, and you should prepare fresh PEG for each experiment
	Low DNA quality	Use an animal transfection grade of DNA purification kit
Decay of FRET signal during imaging	Photobleaching by the excitation	Minimize exposure time and duration of laser excitation