Biotechnology Bulletin ›› 2020, Vol. 36 ›› Issue (1): 1-8.doi: 10.13560/j.cnki.biotech.bull.1985.2019-0550

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Cloning and Expression Profile Analysis of NaERF1 Under Abiotic Stresses in Nicotiana alata

LU Lin1, ZHAO Xi-sheng2, LIU Gui-yun2, LIU Wei-dong1, WANG Yan-ting1, WU Ling-li1, REN Xue-liang3, LU Li-ming2   

  1. 1. College of Landscape Architecture,Sichuan Agricultural University,Chengdu 611130;
    2. College of Agronomy,Sichuan Agricultural University,Chengdu 611130;
    3. Guizhou Academy of Tobacco Sciences,Guiyang 550081
  • Received:2019-06-21 Online:2020-01-26 Published:2020-01-08

Abstract: AP2/ERF transcription factors,unique in plants and the largest family of transcription factors in plants,play an important role in plant growth and development. Exploring the physiological function of ERF transcription factors in Nicotiana alata provides useful references for the study of molecular mechanism of N. alata in resistance to abiotic stresses. Homologous cloning method was used for gene cloning,and qPCR was conducted to analyze the expression profile of NaERF1 under abiotic stress. NaERF1 belonging to ERF family was successfully cloned with ORF 819 bp,encoding 272 amino acids. Bioinformatics analysis showed that the protein encoded by NaERF1 had a molecular weight of 30.7 kD and an isoelectric point of 6.07. It had a typical conservative domain of AP2/ERF transcription factor family. NaERF1 was mainly located in the cytoplasm and contained multiple phosphorylation sites. The results of homology analysis showed that NaERF1 gene had high homology with ERF gene of Solanaceae plants,and had the closest relationship with ERF of common tobacco(Nicotiana tabacum). The expression of NaERF1 was tissue-specific,with the highest expression in flowers,the second in stems,and the lower expression in roots and leaves. Meanwhile,the expression of NaERF1presented 5 patterns under abiotic stresses such as high salinity,drought,low temperature,ABA,low potassium and H2O2. Among them,the response to low potassium and ABA stress was strong. The results of this study suggest that NaERF1 gene belongs to AP2/ERF transcription factor family and may be widely involved in abiotic stress response in N. alata.

Key words: Nicotiana alata, transcription factor, ERF, bioinformatics, gene cloning, abiotic stress