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The molecular cloning and functional characterization of ChNAC1, a NAC transcription factor in Cerasus humilis

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Abstract

Plant-specific NAM, ATAF, and CUC (NAC) transcription factors (TFs) play clear roles in plant development and abiotic stress responses. Chinese dwarf cherry (Cerasus humilis) is an economically important shrub, that has strong resistance to drought. In this study, we isolated and functionally characterized a novel NAC TF from C. humilis. The ChNAC1 ORF contained 894 nucleotides, encoding 297 amino acid residues. ChNAC1 amino acid sequences had the highest similarity with homologous petunia (Petunia hybrida) and tomato (Solanum lycopersicum) NAM proteins. The ChNAC1 transcripts were most abundant in the leaves of seedlings and significantly up-regulated by drought stress. The nuclear localization and transcriptional activity of the C-terminal domain further confirmed that ChNAC1 functions as a TF. Yeast two-hybrid results showed that ChNAC1 can homodimerize in yeast cells. Next, we transformed ChNAC1 into wild-type Arabidopsis thaliana and found that the ectopic expression of ChNAC1 increased chlorophyll, water, proline, and protein contents as well as higher peroxidase (POD) and superoxide dismutase (SOD) activities while decreasing electrolyte conductivity and reactive oxygen species (ROS) contents compared to wild-type and mutant lines. Further, overexpression of ChNAC1 positively regulated ABA-responsive genes under drought stress and increased ABA sensitivity during root growth. Collectively, these results demonstrate the key role of ChNAC1 in drought stress tolerance.

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Abbreviations

WT:

Wild type

OX:

Overexpression

PCR:

Polymerase chain reaction

RT-PCR:

Reverse transcription PCR

qRT-PCR:

Quantitative real-time PCR

RW3:

Rewater 3 days

GFP:

Green fluorescent protein

His:

Histidine

Trp:

Tryptophan

Leu:

Leucine

ABA:

Abscisic acid

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Acknowledgements

This work was supported by Fundamental Research Funds for the Central Universities (Grant No. 2572018CG02), the National Natural Science Foundation of China (Grant No. 31170569) and the Innovation Project of State Key Laboratory of Tree Genetics and Breeding (Northeast Forestry University).

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Authors and Affiliations

  1. State Key Laboratory of Tree Genetics and Breeding (Northeast Forestry University), Harbin, 150040, People’s Republic of China

    Xing Shun Song

  2. Department of Genetics, College of Life Science, Northeast Forestry University, Harbin, 150040, People’s Republic of China

    Fang Wang, Jie Wan Wang, Li Jiao Sun & Xing Shun Song

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  1. Fang Wang
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  2. Jie Wan Wang
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  3. Li Jiao Sun
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  4. Xing Shun Song
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Contributions

XSS conceived and designed the study. FW carried out the main experiments and wrote the manuscript. JWW cloned the ChNAC1 gene. LJS performed the gene expression analysis. All authors discussed the results and commented on the manuscript.

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Correspondence to Xing Shun Song.

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Wang, F., Wang, J.W., Sun, L.J. et al. The molecular cloning and functional characterization of ChNAC1, a NAC transcription factor in Cerasus humilis. Plant Growth Regul 89, 331–343 (2019). https://doi.org/10.1007/s10725-019-00536-9

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