Abstract
Key message
We report that GhWRKY21, a WRKY transcription factor, plays essential roles in regulating the intensity of the drought-induced ABA signalling pathway by facilitating the expression of GhHAB in cotton (Gossypium hirsutum).
Abstract
Abscisic acid (ABA) is one of the most important plant hormones in response to abiotic stress. However, activation of the ABA signalling pathway often leads to growth inhibition. The mechanisms that regulate the intensity of ABA signals are poorly understood. Here, we isolated and analysed the cotton group IId WRKY transcription factor (TF) gene GhWRKY21. Functional analysis indicated that GhWRKY21 plays a negative role in the drought response of cotton. Silencing of GhWRKY21 in cotton dramatically increased drought tolerance, whereas ectopic GhWRKY21 overexpression in Nicotiana benthamiana decreased drought tolerance. Furthermore, the GhWRKY21-mediated drought tolerance was ABA dependent. To clarify the mechanism underlying the GhWRKY21-mediated regulation of drought tolerance, 17 clade-A-type type 2C protein phosphatase (PP2C) genes, which are negative regulators of ABA signalling, were identified in cotton. Notably, GhWRKY21 interacted specifically with the W-box element within the promoter of GhHAB and regulated its expression. Silencing of GhHAB in cotton yielded a phenotype similar to that of GhWRKY21-silenced cotton. These results suggest that GhWRKY21 regulates the intensity of ABA signals by facilitating the expression of GhHAB. In summary, these findings dramatically improve our understanding of the function of WRKY TFs and provide insights into the mechanism of ABA-mediated drought tolerance.
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Acknowledgements
This work was financially supported by grants from the National Natural Science Foundation of China (Grant Nos. 31901431 and 31971823) and the Natural Science Foundation of Shandong Province (Grant No. ZR2019BC015).
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This work was financially supported by grants from the National Natural Science Foundation of China (31901431 to C.W. and 31971823 to X.G.) and the Natural Science Foundation of Shandong Province (ZR2019BC015 to C.W.).
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C.W. and X.G. conceived the project and designed the experiments. X.G. was responsible for the material distribution. J.W., L.W. and Y.Y. performed most of the experiments with the assistance of H.L., Z.G. and S.Z.. J.W. and C.W. wrote the article with the assistance of S.Z.
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Additional file2 (PDF 1322 kb) Fig. S1 Expression of the GhWRKY21-His protein analysed by SDS-PAGE. lane M: protein marker; lane U: recombinant GhWRKY21-His protein without induction; lanes I: recombinant GhWRKY21-His protein with IPTG-induction. Fig. S2 Subcellular localization of GhWRKY21. a Schematic diagram of the control 35S::GFP construct and the 35S::GhWRKY21-GFP fusion protein construct. b Fluorescent signals of the 35S::GhWRKY21-GFP fusion construct and the 35S::GFP construct in N. benthamiana leaves at 2 days after transformation. Green fluorescence was observed with an LSM 880 META confocal microscope (Carl Zeiss). Fig. S3 Expression of GhWRKY21 in T2 Vec and OE plants as measured by qRT-PCR-based analysis. The different letters indicate significant differences (P<0.05) according to Tukey’s honestly significant difference (HSD) test. Fig. S4 Seed germination rates and phenotypes of Vec and OE plants on MS media consisting of different concentrations of mannitol. Fig. S5 Stomatal aperture of Vec and OE plants under drought stress. The data represent the means ± SEs of 40 stomata (length/width) from three independent experiments. The different letters indicate significant differences (P<0.05) according to Tukey’s HSD test. Fig. S6 Expression pattern of GhHAB under ABA treatment. Different letters indicate significant differences (P<0.05) based on Tukey’s HSD test.
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Wang, J., Wang, L., Yan, Y. et al. GhWRKY21 regulates ABA-mediated drought tolerance by fine-tuning the expression of GhHAB in cotton. Plant Cell Rep 40, 2135–2150 (2021). https://doi.org/10.1007/s00299-020-02590-4
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DOI: https://doi.org/10.1007/s00299-020-02590-4