Molecular characterization of two banana ethylene signaling component MaEBFs during fruit ripening

• Two EBF genes designated as MaEBF1 and MaEBF2 were isolated and characterized from banana fruit.
• MaEBF1 and MaEBF2 showed differential expression patterns during fruit ripening.
• Promoter activity of MaEBF2 was activated by ethylene application.
• MaEBF2 was able to physically interact with MaEIL5.

EIN3 Binding F-box protein (EBF) is an essential signaling component necessary for ethylene response. However, little information is available on EBF genes during banana fruit ripening. Two EBF genes designated MaEBF1 and MaEBF2 were isolated and characterized from banana fruit. Subcellular localization analysis showed that MaEBF1 and MaEBF2 were both nuclear proteins. Expression of MaEBF1 and MaEBF2 in fruit with four ripening characteristics revealed that MaEBF2 was enhanced by ethylene during fruit ripening, while MaEBF1 changed only slightly. Moreover, the MaEBF2 promoter was activated after ethylene treatment, further supporting its involvement in fruit ripening. More importantly, MaEBF2 was shown to physically interact with MaEIL5, using yeast two-hybrid (Y2H) and bimolecular fluorescence complementation (BiFC) assays. Together, these results suggest that MaEBF may be involved in banana fruit ripening, at least partly via interaction with MaEIL5. Our findings expand our understanding of the regulatory network of ethylene signaling cascade in banana fruit ripening.