## 2 Results
### Subcellular Comparmentation of Rice PTs in Rice Protoplasts  
To investigate the subcellular localization of rice PTs, synthetic green fluorescent protein (sGFP) was fused to their C-termini and transfected into the leaf protoplastics prepared from 10-day-old rice seedlings (Figure 6). For rice Group I families, green fluorescence for OsGPS:sGFP, OsGGPS1:sGFP and OsGRP:sGFP were similarly observed to be spread in stroma with strong spot-like patterns inside chloroplasts (Figure 6A). Two latter ones were also strongly detected in stroma (Supplementary Figure S3). To verify the subcellular structure of these spot-like patterns, an OsPSY2:mCherry construct, which is targeted into plastoglobules [38], was individually co-transfected. Green fluorescence exactly merged with red fluorescence, indicating that OsGPS, OsGGPS1 and OsGRP proteins localize mainly to both plastoglobules and stroma of chloroplasts. For rice Group II PTs, OsPSPS1:sGFP, OsPS2:sGFP, OsSPSS3:sGFP and OsSPSS4:sGFP were transfected into rice protoplasts and their green fluorescence resulted in different patterns in localization (Figure 6B). OsPS1 was localized in two places of stroma and strong spot-like patterns outside chloroplasts. To verify the cellular Comparmentation of OsPS1, the Red CMXRos fluorescent signals of MitoTracker as a mitochondrial marker probe were presented as yellow signals. These signals matched with green fluorescence of OsSPSS1:sGFP, indicating the dual localization to stroma and mitochondria. OsPSPS2:sGFP and OsSPSS3:sGFP were localized to stroma of chloroplasts. The green fluorescence for OsPSPS4:sGFP was observed as big spot-like patterns, but were still in cytoplasm. To verify these unusual signals, three organelle-specific markers of MitoTracker, G-rb for Golgi apparatus and px-rb for peroxisomes with red fluorescence were co-transfected, but did not merge with the green fluorescent signal of OsSPSS4:sGFP at all, proposing its cytosolic localization into some kinds of vesicles. For rice Group III PTs, five sGFP fusion constructs with OsFS1-5 were individually transfected into rice protoplasts (Figure 6C). OsFPS1-4 nicely showed cytosolic localization patterns, but OsFPS5 was aggregated as a few big-spots, but still in cytoplasm (Figure 6C). Figure 5: Expression profiles of rice _trans_-PTs in response to different hormone treatments in 10-day-old leaves. The heatmap graphs are presented by the relative fold-change expression in comparison to their respective controls. The \(\Delta\)Ct values of each time point (0, 0.5, 1, 2, 4 or 6 h) post treatment (hpt) were calculated by the \(\Delta\)Ct equation against the \(\mathit{OsEF1\alpha}\) gene and the relative fold-change values (2\({}^{-\Delta\Delta\)Ct) of each time point were obtained by the normalization using the \(\Delta\)Ct values of each 0 h time point as a respective control. The 0 h time points of each treatment are presented using the \(\Delta\)Ct values in the gray colored boxes and other time points are displayed using the fold change values. The color key of the heatmap graph is presented under the last graph. The results are expressed as the mean values of three technical repeats and also listed in Table S7. Mock, with no chemical; meJA, methyl jasmonic acid; GA, gibberellic acid; ABA, abscisic acid. Figure 6: Subcellular localization of rice _trans_-PTs. Green fluorescent protein (sGFP) was fused to rice _trans_-PTs of Group I (**A**), Group II (**B**) and Group III (**C**) and sGFP-fused constructs were transfected into rice toptoplasts of 10-day-old leaves. The images were acquired at 630x magnification using a confocal microscope. The white signals of Merged-1 are derived from overlap of the greenGFP-fluorescence signal and the blue chlorophyll autofluorescence. The mCherry fused construct of _O.