TaUBC24 interacted physically with TaICE1
According to the high ICE1 content of OEmiR399 transgenic Arabidopsis
and CsUBC24 interacting with CsICE1 in previous study (R. Wang et al.,
2020), we supposed TaUBC24 might interact with TaICE1. To confirm the
protein-protein interaction between poplar TaUBC24 and TaICE1, the
full-length CDS of TaICE1 fused to AD (AD- TaICE1) was tested
interaction with the full-length CDS of TaUBC24 fused to BD (BD-
TaUBC24). As shown in Fig. 13A, an
interaction between the TaUBC24 and TaICE1 was detected in the Y2H
assay.
To verify the protein interaction between TaUBC24 and TaICE1 in plant
cell, the bimolecular fluorescence complementation (BiFC) was performed.
Coding sequences of TaUBC24 were fused to the N-terminal fragment
of green fluorescent protein (GFP)
(TaUBC24 -GFPN), while TaICE1 was fused
to the C-terminal fragment of GFP
(TaICE1 -GFPC). GFP fluorescence was detected in
the nucleus of N. benthamiana epidermal cells
co-transfected with plasmids that harbored either theTaUBC24 -GFPN andTaICE1 -GFPC (Fig. 13B). These results showed
that TaUBC24 was capable of interacting with TaICE1 in vivo.