4.4. Overexpression of CmWUS in A. thaliana

CmWUS was amplified using primers CmWUS-F2 and CmWUS-R2 (Supplementary Table S1) and subcloned into NcoI/BstEII-cleaved pCambia1304 vector under the CaMV35S promoter using In-Fusion^® HD Cloning Kit System (Clontech, Mountain View, CA, USA). The resulting pCambia1304-CmWUS vector was transformed into A. thaliana (Columbia) via Agrobacteriaum tumefaciens GV3101with the floral dip method [54]. The seeds were selected on MS medium containing hygromycin B (50 mg/L; Roche, Basel, Switzerland). qRT-PCR was performed using young leaves to confirm positive lines with primers CmWUS-F3/R3 and AtACTIN-F/R (Supplementary Table S1). Three independent homozygous T₃ lines with higher and consistent expression levels were selected for floral phenotype analysis. Forty flowers were analyzed and the significant differences were determined according to Fisher’s LSD (p < 0.05) with SPSS 20.0.