Bimolecular fluorescence complementation (BiFC) assay
The full-length cDNA sequences of CsJAZ6 , CsEGL3 , andCsTTG1 were cloned into the cYFP and nYFP vectors. Six fusion
constructs obtained, including CsJAZ6-YFPN,
CsJAZ6-YFPC, CsEGL3-YFPN,
CsEGL3-YFPC, CsTTG1-YFPN, and
CsTTG1-YFPC, were transformed into Agrobacterium
tumefaciens strain GV3101. CsJAZ6-YFPN was paired
with CsEGL3/CsTTG1-YFPC and
CsJAZ6-YFPC with CsEGL3/CsTTG1-YFPNfor transient coexpression in tobacco leaves. The YFP fluorescence
signal for each combination was imaged at 44-48 h after infiltration
using a Leica TCS SP5 confocal laser scanning microscope system (Leica
Microsystems, Bannockburn, IL, USA). An excitation wavelength of 488 nm
and fluorescence detection at 500-542 nm were used.