Cloning and Expression Analysis of NF-YB3 Gene from Vitis vinifera
The full length of the VvNF-YB3 gene was cloned from the European grape'Thompson Seedless'by PCR,to explore the function of the VvNF-YB3 gene in grape growth and development and response to abiotic stresses.The coding region sequence(CDS)of the VvNF-YB3 gene was 624 bp in length,encoding 207 amino acids,with an isoelectric point of 6.31,an instability index of 27.66,and a total average hydrophobic index of-0.738.Therefore,VvNF-YB3 was an acidic and stable hydrophilic protein.Through bioinformatics analysis,it was found that the VvNF-YB3 gene was located on chromosome 10 of the European grape,containing an intron and a CBFD_NFYB_HMF conserved domain,and showed the highest identity with VrNF-YB3 from Vitis riparia.The prediction result of secondary structure showed that VvNF-YB3 was mainly composed of α-helix and random coil.Correspondingly,the tertiary structure was a monomeric helix-coil-helix.The analysis of the cis-acting elements of the VvNF-YB3 gene promoter suggested that it might respond to hormones and stresses.To prove this inference,its expression patterns under different treatments were analyzed.The VvNF-YB3 gene was expressed in leaves,tendrils,flowers,young fruits,and stems of Red Globe,and the expression level was the highest in mature leaves.PEG simulated drought,low-temperature,and high-temperature stresses could change the expression level of the VvNF-YB3 gene,and VvNF-YB3 was a stress responsive gene.In addition,most of the interacting proteins of VvNF-YB3 were subunits of the nuclear factor Y family,and there were 27 transcription factors such as ERF,bHLH,ARF,etc.in the upstream of the VvNF-YB3 gene.It is speculated that the VvNF-YB3 gene may respond to various environmental stresses by being regulated by upstream transcription factors and interacting with other NF-Y proteins.
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