Abstract
Background
Promoters play key roles in plant gene expression in complex and varied natural environments. The type and amount of cis-acting elements in the promoter sequence tend to indicate the response of genes to induction factors. WRAB18 is a group III member of the late embryogenesis abundant (LEA) protein family that performs multiple functions in plant stress physiology. To elucidate the particularly biological effects of WRAB18 on stress, exploration of its promoter sequence is necessary.
Methods and results
In this study, the full-length and promoter sequences of Wrab18 were isolated from the Zhengyin 1 cultivar of Triticum aestivum. The gene sequences and cis-acting elements in the promoter were analyzed using the Plant Promoter Database and bioinformatics methods. The results showed that Wrab18 possessed one intron with 100 bp, the promoter sequence contained various stress-related cis-acting elements, and the functionality of the promoter was checked using green fluorescent protein (GFP) marker protein expression by transient assay in Nicotiana benthamiana. Furthermore, based on promoter prediction analysis, quantitative real-time fluorescent PCR results confirmed the response of gene expression levels to stress factors.
Conclusions
In summary, the promoter sequence of Wrab18 plays a role in plant stress responses, contains multiple cis-acting elements, and provides insights into the role of WRAB18 in plant resilience to stress. This study has guiding significance for further studies of gene function and mechanism of action, and lays a theoretical foundation for improving wheat quality.
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Change history
03 November 2023
A Correction to this paper has been published: https://doi.org/10.1007/s11033-023-08783-w
Abbreviations
- LEA:
-
Late embryogenesis abundant
- qPCR:
-
Quantitative real-time fluorescent PCR
- GFP:
-
Green fluorescent protein
- TSS:
-
Transcription start site
- CTAB:
-
Cetyltrimethylammonium bromide
- LB:
-
Luria-Bertani
- CerealsDB:
-
Chinese Spring in Cereals Database
- ORF:
-
Open reading frame
- PPD:
-
Plant Promoter Database
- ABA:
-
Abscisic acid
- ABREs:
-
Abscisic acid responsive elements
- GARC:
-
GA response complex
- MBS:
-
MYB binding sites
- MCS:
-
MYC recognition sites
- LTREs:
-
Low temperature-responsive elements
- GARE:
-
GA response element
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Acknowledgements
The authors thank Dr. Zhenqing Bai for his great comments on the earlier version of this manuscript. We also thank Dr. Wenbo Yang for her kind providing for pTF486 vector using in the transient expression assay in N. benthamiana.
Funding
This study was funded by the State Key Laboratory of Crop Stress Biology for Arid Areas (grant number: CSBA2015007) and the Specialized Research Fund for the Doctoral Program of Higher Education (grant number: 20120204110033).
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All authors contributed to the study conception and design. Xiaoyu Wang and Linsheng Zhang designed the study. Zhengyang Yu and Weining Zhu analyzed the promoter sequence, and performed experiment with Xiaoyu Wang and Hao Liu. Xiaoyu Wang prepared the manuscripts, Bo Wang performed the transient expression assay and participated in the revision of the article. All authors read and approved the final manuscript.
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Wang, X., Liu, H., Yu, Z. et al. Characterization of wheat Wrab18 gene promoter and expression analysis under abiotic stress. Mol Biol Rep 50, 5777–5789 (2023). https://doi.org/10.1007/s11033-023-08485-3
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DOI: https://doi.org/10.1007/s11033-023-08485-3