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A novel cis-acting element in the GmERF3 promoter contributes to inducible gene expression in soybean and tobacco after wounding

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Using in silico and functional analyses, we cloned and validated the expression profile of an inducible soybean promoter (GmERF3) along with its novel wound-induced and delayed expression (WIDE) element.

Abstract

Promoters and their contributing promoter elements are the main regulators of gene expression at the transcriptional level. Although the Ethylene Response Factor (ERF) gene family is one of the most well-studied stress-responsive gene families in plants, their promoter regions have received little attention. In this study, we investigated the expression patterns driven by the soybean (Glycine max) GmERF3 promoter and its cis-acting elements in soybean and tobacco. Transcriptomic data revealed that the native GmERF3 gene was differentially expressed in organs and tissues of plants. In transgenic soybeans containing a 1.3 kb GmERF3 promoter fused to the green fluorescent protein (gfp) gene, organ- and tissue-specificity was observed in untreated plants while mechanical wounding led to induction of GFP expression. Further in silico and in planta analyses of the GmERF3 promoter sequence in soybean revealed different cis-acting elements, including a novel cis-acting element, which contributed to increased expression, 1–2 days after mechanical wounding. We have named this DNA motif the wound-induced and delayed expression element (GGATTCAAGTTTAACC). A synthetic promoter containing a tetrameric repeat of this element showed high but late wound-induced GFP expression in leaves of transgenic tobacco. Our study expands the toolbox of inducible promoters and promoter elements for potential use in basic and applied research.

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Acknowledgments

We would like to thank Drs. Eric Stockinger, Michelle Jones and Anne Dorrance for critical review of this manuscript and constructive suggestions, Dr. Paul Rushton (Texas A&M University) for his advice on use of synthetic promoters, Cheri Nemes and Amanda Miller for technical assistance with plant transformation, and Dr. Lonnie Welch and Xiaoyu (Veronica) Liang (Ohio University) for their advice using WordSeeker. Salaries and research support were provided by State and Federal funds appropriated to The Ohio State University/Ohio Agricultural Research and Development Center. This research was also partially supported by the United Soybean Board and by a fellowship from Consejo Nacional de Ciencia y Tecnología (CONACYT), Mexico, to CMHG. Mention of trademark or proprietary products does not constitute a guarantee or warranty of the product by OSU/OARDC and also does not imply approval to the exclusion of other products that may also be suitable.

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  1. Carlos M. Hernandez-Garcia

    Present address: Epicrop Technologies, Inc., 5701 N 58th St, Suite 1, Lincoln, NE, 68507, USA

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  1. Department of Horticulture and Crop Science, OARDC/The Ohio State University, 1680 Madison Ave., Wooster, OH, 44691, USA

    Carlos M. Hernandez-Garcia & John J. Finer

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  1. Carlos M. Hernandez-Garcia
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Correspondence to John J. Finer.

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Communicated by B. Li.

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Hernandez-Garcia, C.M., Finer, J.J. A novel cis-acting element in the GmERF3 promoter contributes to inducible gene expression in soybean and tobacco after wounding. Plant Cell Rep 35, 303–316 (2016). https://doi.org/10.1007/s00299-015-1885-7

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