Abstract
Drought affects soybean growth and ultimately led to yield reduction. WRKY transcription factors involve in the regulation of abiotic stress. Few functions of WRKY transcription factors underlying drought tolerance in soybean are clear. Here, we reported a WRKY transcription factor named GmWRKY17 that positively regulates soybean drought tolerance by regulating drought-induced genes and ABA-related genes. Transcriptome sequencing (RNA-Seq) and yeast one hybrid analysis identified downstream genes regulated by GmWRKY17. ChIP-qPCR, EMSA and dual-luciferase reporter assay showed that GmWRKY17 directly bound to the promoters of the GmDREB1D and GmABA2, and activated their expression under drought stress. Overexpression of GmDREB1D gene enhanced drought tolerance of soybean. Taken together, our study revealed a regulatory mechanism that GmWRKY17 transcription factor may improve soybean drought tolerance by mediating ABA synthesis and DREB signaling pathway.
Key message
GmWRKY17 improves the drought tolerance of soybean by reducing water loss, promoting ABA synthesis, and activating GmDREB1D and GmABA2 genes.
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Acknowledgements
The G10 gene (selection marker gene) was provided by Prof. Shen Zhicheng (Institute of Insect Science at Zhejiang University, China). Zhongdou 32 [Glycine max (L.) Merr.] was provided by Prof. Zhou Xin’an (Institute of Oil Crops, Chinese Academy of Agricultural Sciences).
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This research was financially supported by the National Transgenic Major Program, China (2019ZX08013001-001 and 2019ZX08013006-001).
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YC and YL conceived and designed the research. YL conducted the experiments. YL wrote the manuscript draft. YC finalized writing and revision of the manuscript. All authors have read and approved the final version of the manuscript. The authors declare no competing interests.
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Fig. S1
. The expression level of soybean WRKY genes and overexpression of GmWRKY17 gene enhance drought tolerance in Arabidopsis thaliana. Fig. S2. Genetic transformation of soybean and molecular identification of transgenic soybean. Fig. S3. Plant height of RNAi-interfered plants. Fig. S4. Plant height of GmWRKY17-overexpressing plants. Fig. S5. Stomatal densities of WT, GmWRKY17-overexpressing plants, and RNAi plants. Fig. S6. Overexpression of GmWRKY17 effects on agronomic trial under drought stress. Fig. S7. The phenotype of ZD32 and GmWRKY17ox-18 plants under normal growth and drought conditions. Fig. S8. Expression levels of stress-induced genes in WT, OX-13 and OX-18 plants. Fig. S9. Yeast one-hybrid analysis, using pGADT7-GmWRKY17 as the prey, ProGmNCED4-AbAi or ProGmAAO3-1-AbAi as the bait. Fig. S10. Sequence alignment and phylogenetic tree of GmDREB1D protein. Fig. S11. Phenotypes of soybean overexpressing the GmDREB1D gene. Fig. S12. Potential phosphorylation sites of GmWRKY17 protein were predicted by the GPS 6.0 Server. Fig. S13. Detection of Myc-GmWRKY17 protein in soybean protoplasts by Coomassie staining and Western blot. Fig. S14. The phenotype of ZD32, GmWRKY17ox-11 and GmWRKY17ox-18 plants under drought conditions. Table S1. WRKY genes were induced by long-term drought stress. Table S2. Statistical analysis of plant height, pods per plant, 100-seed weight of ZD32 and GmWRKY17-overexpressing plants under normal growth condition and drought stress. Table S3. Genes with increased and decreased induction by drought in ZD32 and GmWRKY17ox-13. Table S4. Primer sequences used in the study. (DOCX 11464 KB)
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Liu, Y., Cao, Y. GmWRKY17-mediated transcriptional regulation of GmDREB1D and GmABA2 controls drought tolerance in soybean. Plant Mol Biol 113, 157–170 (2023). https://doi.org/10.1007/s11103-023-01380-2
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DOI: https://doi.org/10.1007/s11103-023-01380-2