한국 콩(Glycine max (L.) Merr) 품종의 효율적인 원형질체 분리 및 유전자 도입

Efficient Isolation and Gene Transfer of Protoplast in Korean Soybean (Glycine max (L.) Merr.) Cultivars

Soybean (Glycine max (L.) Merr.) is one of the most important crops with economic value as a source of protein and vegetableoil for human food and animal feed. In recent years, rapidly developed genome editing techniques have shown widespread application prospectsfor gene function studies and for improving important agronomic traits in many crops. Therefore, it is important to establish a highly efficientmethod for protoplast isolation and transient expression systems in soybeans. In this study, we established an efficient method for protoplastisolation and its application to transient gene expression in Korean soybean cultivars. The protoplasts were isolated from leaves, epicotyls,hypocotyls, cotyledons, and etiolated hypocotyls using various combinations of enzyme mixtures. We found that high-quality and large amountsof protoplasts were isolated from the etiolated hypocotyls when incubated for 8 h under conditions of 0.5% cellulase, 0.5% pectinase, and1% viscozyme. In addition, we observed a high transfection efficiency of green fluorescent protein using etiolated hypocotyl protoplasts. Takentogether, our protoplast isolation and transfection method is highly efficient and can be used for gene function and molecular analysis tobetter understand the biological and physiological processes in soybean.