Molecular cloning, transcriptional activation, subcellular localization analysis and expression characterization of ZjERF1 from Zoysia japonica

doi:10.11686/cyxb2018323

Abstract

Abstract: The ERF transcription factors (TFs) are one of the largest groups in plants and play important roles in plant developmental progress and responses to abiotic stresses. In this study, one novel ERF gene, ZjERF1 (GenBank No. MH294481), was isolated from Zoysia japonica using the RACE method. The open reading frame of ZjERF1 is 630 bp in length, encoding 209 amino acids. One highly conserved AP2 domain was found in ZjERF1, indicating ZjERF1 was a typical ERF TF. By genome walking, a 1581 bp upstream sequence from the ATG of ZjERF1 was obtained. Bioinformatics analysis revealed that there were several cis-elements in response to MeJA and abiotic stresses in the promoter. To investigate the transcriptional activation character, a pGBKT7-ZjERF1 vector was constructed and then was transformed into Y2HGold yeast cells; the results showed that ZjERF1 had strong transcriptional activity. To reveal the subcellular localization character, 35S::ZjERF1:YFP was generated. The transient expression analysis in Nicotiana benthamiana demonstrated that ZjERF1 was localized in the nucleus. To further explore the expression characteristics, real-time quantitative PCR was carried out. The results showed that ZjERF1 was expressed most abundantly in the stem and its transcriptional abundance was positively correlated with leaf senescence. Moreover, its expression could be induced by 200 μmol·L^-1 ET, 10 μmol·L^-1 MeJA or 300 mmol·L^-1 NaCl but was suppressed by 20% PEG4000. Taken together, these findings proves that ZjERF1 is a functional TF involved in various signaling pathways and paves the way for further study of the function of ZjERF1 and its regulatory mechanism in Z. japonica.

Key words: Zoysia japonica, ERF transcription factor, transcriptional activity, subcellular localization, expression character

TENG Ke, ZHANG Rui, TAN Peng-hui, YUE Yue-sen, FAN Xi-feng, WU Ju-ying. Molecular cloning, transcriptional activation, subcellular localization analysis and expression characterization of ZjERF1 from Zoysia japonica[J]. Acta Prataculturae Sinica, 2019, 28(6): 56-65.

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