Characteristic and Expression Analysis of a Transcription Factor PsZFP1 in Tree Peony

doi:10.7668/hbnxb.20191827

Abstract

Abstract: Zinc finger proteins(ZFPs), which containing conserved zinc finger domains, play important roles in the plant growth and development as well as in stress responses. In order to further explore the function and mechanism of PsZFP1 in the process of breaking dormancy in tree peony, the full-length cDNA sequence of PsZFP1 was cloned from Paeonia suffruticosa Luhehong by the 5'-RACE. After that, the amino acid sequence and phylogenetic tree of PsZFP1 were analyzed by the EditSeq and MEGA 6.0 software. Then, Agrobacterium tumefaciens-mediated transient transformation of tobacco was used to investigate subcellular localization of PsZFP1, and the pGBKT7-PsZFP1 recombinant plasmid was constructed for the transcriptional activity analysis. Yeast one hybridization was used to analysis the interaction between PsZFP1 and PsMPT promoter. In addition, the qRT-PCR was performed for the expression analysis of PsZFP1 in the flower buds of tree peony along with chilling duration process. The pET28a⁺-PsZFP1 recombinant plasmid was also constructed and expressed in BL21 in order to obtain PsZFP1 protein. The results showed that the PsZFP1 cDNA was 1 313 bp, and its open reading maximum frame was 915 bp, which encoded a protein with 304 amino acid. The PsZFP1 protein was a hydrophilic and stable protein, which contained a conserved CCCH domain, and shared a high degree of sequence similarity with a VvZFP from grape. The PsZFP1 protein was located in the nucleus, and was a transcriptional factor which had transcriptional activation activity. Further yeast one hybridization analysis confirmed that PsZFP1 could bind to the MYC element of PsMPT promoter. In addition, the expression of PsZFP1 gene increased in the flower buds of tree peony along with chilling duration process, and the highest expression of PsZFP1 gene was presented at 21 d chilling and decreased significantly at 28 d chilling. Moreover, the PsZFP1 protein was expressed in prokaryotic cells, and the purified protein PsZFP1 was then obtained by Ni-NTA affinity chromatography and detected by SDS-PAGE. Taken together, we identified a zinc finger transcriptional factor, which could be induced by chilling accumulation and regulate PsMPT expression for energy metabolism during chilling induced dormancy release. The results were benefit to further analyze the function of PsZFP1 and its regulation to endodormancy.

Key words: Tree peony, PsZFP1, Transcription factor, Prokaryotic expression, Chilling

CLC Number:

Q78
S682

LIU Qing, SUN Tingzhao, GAI Shupeng, ZHANG Yuxi, LIU Chunying. Characteristic and Expression Analysis of a Transcription Factor PsZFP1 in Tree Peony[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2021, 36(3): 67-73. doi: 10.7668/hbnxb.20191827.

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