Subcellular Localization of OsGA2ox5 and OsGHD7

The coding region of OsGA2ox5 was amplified using the primer pair OsGA2ox5-pA7YFPF and OsGA2ox5-pA7YFPR (sangon) (the specific primers are listed in supplemental Table S1) and cloned into pA7-YFP [31] (link), generating the OsGA2ox5-YFP fusion under the control of the CaMV 35S promoter. A previously study demonstrated that OsGHD7 [32] (link) is a nuclei protein and localized in nuclei only, so we used OsGHD7 as a positive control. The OsGHD7 coding sequence was fused in frame to the N-terminus of YFP under the control of the CaMV 35S promoter. Then, OsGA2ox5-YFP, OsGHD7-YFP fused construct and pA7-YFP vectors were used to transiently transform onion epidermal cells by particle bombardment[33] using a particle gun system (PDS-1000/He; Bio-Rad). After 24 h, the epidermal cells were examined for YFP fluorescence under a scanning confocal microscope (Zeiss LSM510; Carl Zeiss Micro-Imaging GmbH, Jena, Germany).