2.4 Subcellular localization
The full length coding sequence of OsLHY was fused to green fluorescent protein (GFP) in the pM999 vector under the control of the CaMV 35S promoter. Meanwhile, the RPL1 coding sequencing was used to fuse with cyan fluorescent protein (CFP), as RPL1 can be used as a nuclear marker (Zhang, Yuan, & Zhang, 2012). Then the two plasmids were co-transformed into the rice etiolated stems protoplasts isolated from ten-day-old Zhong Hua11, which was grown in the growth chamber at 28 °C under dark conditions. The method for rice protoplasts transformation was based on the previously study (Bart, Chern, Park, Bartley, & Ronald, 2006). Fluorescence signals were detected by an Olympus FluoView FV1000 confocal microscope.