Cloning and Expression Analysis of PtERF98 Gene from Pueraria thomsonii Benth.
The ethylene responsive factors(ERF)are plant-specific transcription factors which play cru-cial regulatory roles in ethylene signal transduction pathways,plant growth and development,as well as stress responses.In order to investigate the expression and regulation mechanism of the PtERF98 transcription factor during the post-harvest physiological deterioration(PPD)process in roots of Pueraria thomsonii Benth.,the PtERF98 gene was cloned from the leaf tissue of P.thomsonii Volcano cultivar using RT-PCR technology.The bioinformatics,tissue expression specificity and expression characteristics during the PPD process of PtERF98 were analyzed.The results showed that the PtERF98 gene had a CDS sequence of 450 bp,encoding 149 amino acid residues.The PtERF98 protein had theoretical molecular weight and isoelectric point of 16.93 kDa and 8.06,respectively,and was a hydrophilic unstable protein.Protein structure analysis indicated that PtERF98 contained a typical AP2 conserved domain,so it was categorized as a member of the ERF transcription factor family;it was localized in the nucleus without transmembrane helix structure,but contained one signal peptide and 16 phosphorylation sites.Amino acid sequence homology and the phylogenetic analysis indicated that PtERF98 had the highest homology and the closest phylogenetic relationship with GmERF98 of soybean(Gly-cine max).Tissue expression analysis demonstrated that PtERF98 gene was expressed in the roots,stems and leaves of Volcano cultivar.There were differences in expression level among different tissues,and the highest expression level exhibited in leaves.Moreover,PtERF98 expression was significantly induced by PPD,and its expression level first increased and then decreased over time,reaching the peak at 48 hours post-harvest,which suggested that this gene may be involved in the PPD process of P.thomsonii tuberous roots.This study could lay the groundwork for further understanding the function of PtERF98 gene and its regulatory roles in the PPD process of P.thomsonii.
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