Transient expression in N. benthamiana and confocal laser scanning microscopy
The preparation of Agrobacterium  and transient expression in N. benthamiana  was performed as previously described (Li, 2011).  The vector pB7FWG2:35S:TaA6b-GFP was transformed into A. tumefaciens  GV3101. A. tumefaciens  GV3101 harboring the binary vector was grown in LB-broth media containing 50 mg/L of spectinomycin, 30 mg/L gentamycin, 30 mg/L of rifampin to the stationary phase at 28 °C. After centrifugation, A. tumefaciens  was resuspended in the infiltration medium (10 mM MgCl2, 100 μM acetosyringone) to a final OD600 of 0.5. The suspension was infiltrated with a 1-mL needleless syringe into the abaxial surface of 4-week-old leaves of N. benthamiana . Protein localization was analyzed 24-48 hours after infiltration by confocal laser scanning microscope (TCS SP8, Leica). GFP was excited at 488 nm and the fluorescence detected between 498-525 nm.