Subcellular localization and transcriptional activity analysis of SmGRAS5
To determine the subcellular localization of SmGRAS5, SmGRAS5 protein was fused with a green fluorescent protein (GFP) label. The protoplasts of tobacco leaves and onion epidermal cells were used to analyze the SmGRA5-GFP signal (Fig. 2). The results showed that the GFP fluorescence of SmGRAS5 was enriched in the nucleus, indicating that SmGRAS5 might function as a TF.
To further identify the characteristics of SmGRAS5, the transcriptional activity of SmGRAS5 was analyzed. The results showed that the yeast withSmGRAS5-pGBKT7 and control constructs were able to grow on the SD/-Trp medium. However, the yeast with SmGRAS5-pGBKT7 and negative control constructs could not survive on the SD/-Trp-His-Ade medium (Fig. S2). The results demonstrated that SmGRAS5 had no transcriptional activity.