*4.5. Co-Localization of GmWRKY*

Seeds of Kenong199 were cultivated in a 10 × 10 cm flowerpot (vermiculite: nutritious soil is 1:3), fresh leaf tissue of 7-day-old wheat seedlings were used for preparation of wheat protoplasts. Amplified cDNA sequence of *GmWRKY12* was cloned into the N-terminus hGFP protein driven by the CaMV35S promoter. The cDNA coding sequences of AT2G03340 (AtWRKY3) which located in the nucleus [67] were fused to the N-terminus of the mCherry protein (WRKY25-RFP) under the control of the CaMV 35S promoter [68]. The recombinant plasmid of GmWRKY12-GFP and AtWRKY3-mCherry were co-transformed into wheat mesophyll protoplasts via the PEG4000-mediated method. The 35S::GFP vector was transformed as the control. Fluorescence was observed using a confocal laser scanning microscope (LSM700; CarlZeiss, Oberkochen, Germany) after incubating in darkness at 22 ◦C for 18–20 h. Primers are available in Table S4.
