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Heterologous overexpression of the Arabidopsis SnRK2.8 gene enhances drought and salt tolerance in Populus × euramericana cv ‘Nanlin895’

  • Hui Wei
  • Ali Movahedi
  • Chen Xu
  • Pu Wang
  • Weibo Sun
  • Tongming Yin
  • Qiang ZhugeEmail author
Original Article
  • 43 Downloads

Abstract

The SnRK (sucrose non-fermenting-1 related protein kinase) families respond to salt tolerance, nutritional stress, disease stress, and carbon metabolic regulation. In this study, the SnRK2.8 gene was cloned from Arabidopsis thaliana and nominated AtSnRK2.8. Bioinformatics characterization of AtSnRK2.8 showed that it contained an ATP binding site and a serine/threonine protein kinase binding site at the N-terminal catalytic domain. In addition, the AtSnRK2.8 protein contained special domain I at the C-terminal catalytic domain. Real-time polymerase chain reaction (PCR) results showed that the AtSnRK2.8 gene was expressed in all tested tissues and was highly expressed in seeds. Our analyses proved that the AtSnRK2.8 gene expression was upregulated by salt and drought stresses. The recombinant AtSnRK2.8 protein was expressed using a pET-28a system and purified with HisTrap HP affinity columns. The recombinant AtSnRK2.8 protein has the properties of a protein kinase. The results of overexpressing AtSnRK2.8 in Nanlin895 poplar showed that the transgenic poplar plants had a better ability to resist drought and salt stress compared to the wild type, and adapted well to drought and a high-salt environment.

Keywords

Arabidopsis thaliana Drought and salt Tolerance Heterologous Overexpression Populus SnRK 

Notes

Acknowledgements

This work was supported by the National Key Program on Transgenic Research (2018ZX08020002), the National Science Foundation of China (no. 31570650) and the Priority Academic Program Development of Jiangsu Higher Education Institutions.

Author contributions

HW and AM designed the experiments and wrote the manuscript. CX, PW, WS did help to the HW and AM to do experiments. TY and QZ supervised this project.

Compliance with ethical standards

Conflict of interest

The authors declare no conflicts of interest.

Supplementary material

11816_2019_531_MOESM1_ESM.jpg (66 kb)
Supplementary material 1 Figure S1: Phosphorylation site analysis of the AtSnRK2.8 protein using the SignalP 4.1 Server (http://www.cbs.dtu.dk/services/SignalP/) online program (JPG 65 KB)
11816_2019_531_MOESM2_ESM.jpg (217 kb)
Supplementary material 2 Figure S2: 1 Expression analysis of the AtSRK2.8 gene in transgenic poplar plants. Total RNA was isolated from the leaves of wild-type and transgenic (numbers represent lines) poplar plants, and subjected to RT-PCR analysis. (A) Expression of AtSRK2.8 in AtSRK2.8 transgenic poplar plants. (B) Expression of Ptactin as the internal control in AtSRK2.8 transgenic plants. (C) Quantitative RT-PCR analysis of transcript levels of AtSRK2.8 in AtSRK2.8 transgenic poplar plants (JPG 217 KB)
11816_2019_531_MOESM3_ESM.jpg (772 kb)
Supplementary material 3 Figure S3: Transplanting process for the transgenic poplar after propagation (JPG 771 KB)
11816_2019_531_MOESM4_ESM.jpg (639 kb)
Supplementary material 4 Figure S4: Phenotypic analysis of the wild-type (WT) and AtSRK2.8 transgenic plants before and after the drought and salt treatments. (A) Phenotypic analysis of WT and AtSRK2.8 transgenic plants after a 2-week salt treatment. (B D) Phenotypic analysis of WT and AtSRK2.8 transgenic plants before stress treatment (under normal conditions). (C) Phenotypic analysis of WT and AtSRK2.8 transgenic plants after a 2-week drought treatment. Three independent experiments were performed (JPG 639 KB)

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Copyright information

© Korean Society for Plant Biotechnology 2019

Authors and Affiliations

  1. 1.Co-Innovation Center for Sustainable Forestry in Southern China, Key Laboratory of Forest Genetics and Biotechnology, Ministry of EductionNanjing Forestry UniversityNanjingChina
  2. 2.Jiangsu Provincial Key Construction Laboratory of Special Biomass Resource UtilizationNanjing Xiaozhuang UniversityNanjingChina

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