Improved salt tolerance in transgenic tobacco by over-expression of poplar NAC13 gene
- Published
- Accepted
- Subject Areas
- Genetics, Environmental Impacts, Forestry
- Keywords
- gene over-expression, transcription factor, transgenic tobacco, NAC, salt tolerance
- Copyright
- © 2019 Zhang et al.
- Licence
- This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, reproduction and adaptation in any medium and for any purpose provided that it is properly attributed. For attribution, the original author(s), title, publication source (PeerJ Preprints) and either DOI or URL of the article must be cited.
- Cite this article
- 2019. Improved salt tolerance in transgenic tobacco by over-expression of poplar NAC13 gene. PeerJ Preprints 7:e27861v1 https://doi.org/10.7287/peerj.preprints.27861v1
Abstract
Background: NACs are one of the major transcription factor families in plants which play an important role in plant growth and development, as well as in adverse stress responses.
Methods: In this study, we cloned a salt-inducible NAC transcription factor gene (NAC13) from a poplar variety 84K, followed by transforming it into both tobacco and Arabidopsis.
Results: Stable expression analysis of 35S::NAC13-GFP fusion protein in Arabidopsis indicated that NAC13 was localized to the nucleus. We also obtained five transgenic tobacco lines. Evidence from morphological and physiological characterization and salt treatment analyses indicated that the transgenic tobacco enhanced salt tolerance, suggesting that NAC13 gene may function as a positive regulator in tobacco responses to salt stress. Furthermore, evidence from yeast two-hybrid screening demonstrated that NAC13 protein functions as a transcriptional activator, with an activation domain located in the C-terminal region.
Discussion: NAC13 gene plays an important role in response to salt stress in tobacco. Future studies are needed to shed light on molecular mechanisms of gene regulation and gene networks related to NAC13 gene in response to salt stress, which will provide a valuable theoretical basis for forest genetic breeding and resistant breeding.
Author Comment
In our study, we firstly chose NAC13 gene which belongs to NAC transcription factor in the 84K poplar. Then we constructed NAC13-GFP confusion and confirmed NAC13 protein was targeted in nucleus according to stable expression analysis of transgenic Arabidopsis. Further, evidence from yeast two-hybrid screening demonstrated that NAC13 protein functions as a transcriptional activator, with an activation domain located in the C-terminal region. Finally, we obtained NAC13-overexpressing transgenic tobacco plants and proved that the gene can improve the salt tolerance of plants.
Supplemental Information
Prime names and sequences
The primers of NAC13 are used to amplify the coding sequence of NAC13 gene. NAC13F1 and NAC13R1 are used to clone the ORF sequence of NAC13 gene; NAC13F2 and NAC13R2 are used to generate the recombinant construct 35S::NAC13-GFP, and the underlined area is the restriction site Spe I; NAC13F3 and NAC13R3, NAC13aF and NAC13aR, NAC13bF and NAC13bR are used to generate bait vector pGBKT7-NAC13, pGBKT7-NAC13a and pGBKT7-NAC13b, respectively. The underlined areas are EcoR I and Sal I restriction sites; NAC13F4 and NAC13R4 are used to amplify the complete cDNA sequence of NAC13 gene and introduce into the plant binary vector pBI121. The underlined areas are restriction sites Xba I and Sac I.
The characteristics of NAC13 gene in 84K poplar
Supplemental Figure 1. The characteristics of NAC13 gene in 84K poplar. (A) Gene structure of NAC13 as predicted by Gene Structure Display Sever; (B) Phylogenetic tree analysis of NACs from different plants by MEGA 5.1 using Neighbor-Joining method; (C) Alignment of amino acid sequences of NACs by Clustal W.