Subcellular localization was performed using the Dong et al. (2021) (link). Primers (FveARF2-GFP-F and FveARF2-GFP-R) were designed to amplify the FveARF2 CDS region without a termination codon (Table S1). The LA Taq polymerase (Takara, Dalian, China) was used for PCR, which was performed according to the following procedure: (1) 95 C for 5 min, (2) 95°C for 30 s, 55°C for 30 s, and 72°C for 2 min 30 s for 35 cycles, and (3) 72°C for 10 min. XbaI and BamHI were used to digest the pRI101-GFP vector and 35S::FveARF2-GFP (FveARF2-GFP) was constructed using T4 DNA ligase (Takara, Dalian, China). The pRI101-GFP vector containing 35S::GFP (GFP) was used as a control. FveARF2-GFP and pRI101-GFP were transformed into the Agrobacterium strain GV3101, and the bacterial liquid culture was used to inoculate N. benthamiana leaves. A laser confocal fluorescence microscope (TCS SP8-SE; Leica, Wechsler, Germany) was used to examine GFP fluorescence at an excitation wavelength of 488 nm and an emission wavelength of 505–530 nm.
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