Wheat Protoplast Transient Expression

The full-length coding DNA sequences (CDS) of RCC1-3A (TraesCS3A02G362800), RCC1-3B (TraesCS3B02G395200), RCC1-3D (TraesCS3D02G356500), Myb-7B (TraesCS7B02G188000), and Myb-7D (TraesCS7D02G295400) were inserted into pCambia1300-35S-GFP, creating RCC1-3A::GFP, RCC1-3B::GFP, RCC1-3D::GFP, Myb-7B::GFP, and Myb-7D::GFP fusion vectors. The recombinant plasmids were mixed with the nuclear marker NLS-mCherry and transfected into wheat mesophyll protoplasts as previously described by Yoo et al. (2007) (link). The transfection mixture was induced by PEG-Ca²⁺, and the protoplasts were cultured for 12 h at 25°C. The protoplasts were observed and photographed with a fluorescence microscope (Zeiss Imager A2, Germany).