Molecular Cloning and Characterization of EREB5,a Novel Transcription Factor Gene of ERF Family from Gossypium barbadense L.

In recent years,ERF(Ethylene-responsive element binding factor) family transcription factors have become the research hot spot of crop molecular breeding and molecular mechanisms of resistance or tolerance of plant to bio-or abiotic stresses.In this study,one novel gene,designated as EREB5,was firstly isolated from Gossypium barbadense treated with Verticillum dahliae by cloning in silico,homologic amplification and RACE(Rapid-amplification of cDNA ends) techniques.The bioinformatics analyses demonstrated that the full-length cDNA contained a 573 bp open reading frame which encoded a deduced protein which included 190 amino acid residues,contained an ERF conserved domain,and belonged to the B3 subgroup of ERF subfamily.It also contained one nucleus localization signal,one serine-riched domain,some sites of phospholation and one motif of potato inhibitor I family.Transient expression analysis showed that EREB5 protein was localized in nucleus,meanwhile,the results of Electrophoretic mobility shift assay revealed that EREB5 protein had the ability of binding specifically to the GCC-box.Moreover,the results of expression analysis indicated that EREB5 was able to be induced in Gossypium barbadense seedlings,by inoculating with Verticillum dahliae or stimulation of ethylene,respectively.These results imply that EREB5 should play an important role in resistance of Gossypium barbadense to Verticillum wilt.