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Bulletin of Botanical Research ›› 2018, Vol. 38 ›› Issue (5): 733-740.doi: 10.7525/j.issn.1673-5102.2018.05.013

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Cloning,Construction of Expression Vector and Expression Analysis of the Ethylene-responsive Factor ERF Gene NtTOE3 in Nicotiana tabacum

ZHUO Wei1,2, CHEN Qian1,2, LUO Yin1,2, YANG Shang-Yu1,2, LU Li-Ming1,2, LI Li-Qin1,2   

  1. 1. College of Agronomy, Sichuan Agricultural University, Chengdu 611130;
    2. National Demonstration Center for Experimental Crop Science Education, Chengdu 611130
  • Received:2018-03-30 Online:2018-09-15 Published:2018-08-31
  • Supported by:
    The Foundation of State Key Laboratory of Plant Physiology and Biochemistry,China(SKLPPBKF1505,SKLPPBKF1506)

Abstract: ERF(Ethylene-responsive factor) is a class of ethylene-responsive transcription factor with AP2 signature domain, which plays a pivotal role in response to stress response in plants.To identify the biological functions and regulation mechanism of ERF familymember TOE3 in tobacco k326, one NtTOE3 gene swerecloned from Nicotiana tabacum with homologous cloning strategy, gene expression patterns were analyzed byreal-time quantitative PCR(qRT-PCR), and their functions and regulation mechanisms were predicted bybioinformatic analysis.Theresultsshowed that NtTOE3 contained 1 356 bp and encoded 451 amino acid. The predicted molecular weight was 49.84 ku. By bioinformatics analysis,NtTOE3 was a hydrophilic protein which might be localized in the nucleus,andthe protein had 96% homology with N.sylvestris NsTOE3, thusit was namedas NtTOE3. From expression patterns, the gene was expressed in roots, stems, leaves and flowers in mature stage, which had the highest expression in stems. Expression patterns under abiotic stress indicated the gene responded rapidly to the adverse conditions of low potassium, high salt, drought, H2O2, ABA and 4℃. Therefore, the NtTOE3 transcription factor plays an important regulatory role in tobacco abiotic stress.

Key words: Nicotiana tabacum, NtTOE3, cloning, bioinformatics, gene expression

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