5. Conclusions
In this study, the chloroplast targeting transit peptide and Kz sequence for the enhanced translation efficiency were applied to improve the fluorescence intensity of FCA using the rice protoplast system. The localization of sGFP in chloroplasts contributes to the enlargement of the population with the intensity above the threshold value to increase of transfection ratio on FCA from 14.4% to 64.4%. It suggests that the loss of the transfected population is minimized, which is regarded as a non-transfected population, because of their weak fluorescence intensity similar to the background signals. Also, it plays an important role in the increase of population with high intensity (>104) to improve the average of fluorescence intensity. Our strategy could be usefully applicable to various FCAs using green tissue protoplasts.