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Populus euphratica WRKY1 binds the promoter of PeHA1 to enhance gene expression and salt tolerance

Citation

Yao, Jun et al. (2019), Populus euphratica WRKY1 binds the promoter of PeHA1 to enhance gene expression and salt tolerance, Dryad, Dataset, https://doi.org/10.5061/dryad.83ng085

Abstract

Plasma membrane proton pumps play a crucial role in maintaining ionic homeostasis in salt-resistant Populus euphratica under saline conditions. High levels of NaCl (200 mM) induced PeHA1 expression in P. euphratica roots and leaves. We isolated a 2022-bp promoter fragment upstream of the translational start of PeHA1 from P. euphratica. The promoter-reporter construct PeHA1-pro::GUS was transferred to tobacco plants, demonstrating that β-glucuronidase activities increased in root, leaf, and stem tissues under salt stress. DNA affinity purification sequencing revealed that PeWRKY1 protein targeted the PeHA1 gene. We assessed the salt-induced transcriptional response of PeWRKY1 and its interaction with PeHA1 in P. euphratica. PeWRKY1 binding to the PeHA1 W-box in promoter region was verified by a yeast one-hybrid assay, electrophoretic mobility shift assay, luciferase reporter assay, and virus-induced gene silencing. Transgenic tobacco plants overexpressing PeWRKY1 had improved expression of NtHA4, which has a cis-acting W-box in the regulatory region, and H+ pumping activity in both in vivo and in vitro assays. We conclude that salt stress upregulated PeHA1 transcription due to the binding of PeWRKY1 to the W-box in the promoter region of PeHA1. Thus, we conclude that enhanced H+ pumping activity enabled salt-stressed plants to retain Na+ homeostasis.

Usage Notes

Funding

National Natural Science Foundation of China, Award: 31770643, 31570587

Beijing Natural Science Foundation, Award: 6182030, 6172024

the Fundamental Research Funds for the Central Universities, Award: 2019ZY25

the Program of Introducing Talents of Discipline to Universities, Award: B13007

Location

North China