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Fig. 5 | BMC Plant Biology

Fig. 5

From: Potential function of CbuSPL and gene encoding its interacting protein during flowering in Catalpa bungei

Fig. 5

Interaction between the two proteins CbuSPL9 and CbuHMGA. a Nuclear localization of the CbuSPL9 protein and CbuHMGA protein. The GFP (control) gene, CbuSPL9-GFP fusion gene and CbuHMGA-GFP fusion gene were expressed transiently in Nicotiana benthamiana leaf epidermal cells and observed with confocal microscopy. DAPI, DAPI for nuclear staining image; GFP, GFP green fluorescence image; Merge, the merged images of bright-field, GFP and DAPI staining. b Binding of GST-CbuSPL9 to HIS-CbuHMGA, with GST-CbuSPL9 concentrations of 2381.00 nm, 1191.0 nm, 595.30 nm, 297.60 nm, 148.80 nm assessed by real-time biolayer interferometry. c Yeast two-hybrid assays for the interactions between CbuSPL9 and CbuHMGA. CbuHMGA (as prey) was fused with the GAL4 activation domain (AD) in pGADT7, while CbuSPL9 (as bait) was fused with the GAL DNA-binding domain (BD) in pGBKT7. The positive control was as follows: pGADT7-T and pGBK-53. The negative control was as follows: pGADT7-T and pGBK-Lam. Interactions are indicated by the blue color on SD/−Trp/−Leu/−His/−Ade/X-α-gal medium. d Confocal images of the BiFC analysis in protoplasts from P. trichocarpa. CbuSPL9 was fused with EYFPC, and CbuHMGA was fused with EYFPN. EYFP signal was detected in the nucleus of the S1–21 protoplasts transfected by H2A:mCherry and CbuSPL9:EYFPC with (A) CbuHMGA:EYFPN. Bars = 50 μm

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