*2.5. Subcellular Localization and Expression Patterns of OsCAF1*

OsCAF1 was localized in the chloroplasts, which was predicted using the TargetP website (http://www.cbs.dtu.dk/services/TargetP/). To determine the actual subcellular localization of OsCAF1, we fused the OsCAF1 full-length cDNA with GFP driven by the 2× CaMV 35S promoter and transiently transfected in rice protoplasts. The results showed that the green fluorescent signals of OsCAF1–GFP co-localized with chloroplast autofluorescence in transformed rice protoplasts (Figure 6A). The assays indicated that OsCAF1 was localized in the rice chloroplast.

The qRT-PCR assays were used to investigate the expression patterns of OsCAF1 in different tissues at the three leaves stage. According to our results, OsCAF1 was highly expressed in green tissue, particularly in the leaves, suggesting that the OsCAF1 mainly functions in green tissue (Figure 6B).

**Figure 6.** Subcellular localization and expression patterns of OsCAF1. (**A**) Subcellular localization of OsCAF1 in rice protoplasts. (**B**) Expression analysis of OsCAF1 in different tissues. The tissues include roots (R), stems (S), and leaves (L). The values in the figures represent the means ± SE (*n* = 3). \*\* indicate *p* < 0.01.
