2.13 Subcellular localization analysis of CsTPS1 and CsTPS1-AS
proteins
Subcellular localization assays of CsTPS1 and CsTPS1-AS proteins were
performed following the procedure described in a previous study (Wang et
al., 2019a). Briefly, binary vectors (pCHNP-eYFP/mCherry) were
constructed with several elements on the pCAMBIA1300 backbone (CAMBIA,
Canberra, Australia). The amplified fragments were introduced into
pCHNP-eYFP with the NcoI site using in-fusion technology. The empty
vector pCHNP-mCherry was used as a negative control. Agrobacterium
tumefaciens strain GV3101 carrying the construct for the transient
expression of individual mCherry and CsTPS1 EYFP and CsTPS1-AS EYFP
fusion proteins was mixed and infiltrated into the leaves of tobacco.
Images were taken using a laser confocal fluorescent microscope (Lecia
DMi8, Germany). The EYFP, mCherry fluorescence, and chloroplast
autofluorescence were analyzed at excitation wavelengths of 488 nm, 561
nm, and 561 nm and emission wavelengths of 500–530 nm, 580–620 nm, and
680–720 nm, respectively.