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Cloning of a new LEA1 gene promoter from soybean and functional analysis in transgenic tobacco

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Abstract

LEA1 gene from Glycine max can be expressed in late-embryo stage of plants, and respond to salinity and dehydration stress. To elucidate the mechanism for stress tolerance and high expression in seeds, we isolated and characterized the promoter of LEA1 gene (EQ, 1997 bp) starting the 5′LEA1 coding region. A deletion mutant of EQ promoter (ED) and the full length promoter (EQ) were fused to GUS reporter gene and transformed into the tobacco leaf discs. The results indicated that expression of the reporter gene (GUS) could be regulated by EQ promoter, and was stronger than the mutant under the stress conditions. Also, the expression level of GUS gene driven by EQ promoter in transgenic tobacco seeds was significantly higher than that by the mutant promoter, which meant that it had a better tissue-specificity. Therefore, the active domain for the promoter was located between −1997 and −1000 bp. Additionally, the activity of EQ promoter was 2.1-, 3.3- and 0.4- times stronger than the activity of promoter CaMV35S under salt (24 h), drought (10 h) or ABA (24 h), respectively. Meanwhile, the GUS activity of EQ promoter in seeds was 1.8-fold stronger compared to the promoter CaMV35S. In summary, the new promoter (EQ) is bi-functional, stress-inducible and seed-specific. These findings provide a further understanding for the regulation of LEA1gene expression, and suggest a new way for improving seed quality under saline and alkaline land.

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Acknowledgements

This work was supported by Natural Science Foundation of Heilongjiang province (C201458), Government talent training program (UNPYSCT-2016090), National Nature Science Foundation (31570207), Natural Science Foundation of Heilongjiang province (ZD201408) and National Nature Science Foundation (31301335).

Author contributions

Conceived and designed the overall study -YZ, WS, YW; performed experiments -YZ, YW, LQ, YZ, YZ, MJZ; analyzed the data -YZ, WS, YZ; wrote the manuscript -YZ, WS, LQ, YZ.

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Authors and Affiliations

  1. College of Life Science and Agroforestry, Qiqihaer University, Qiqihaer, 161006, Heilongjiang, China

    Yan Zhao, Ying Zhai, Meijuan Zhang & Wei Sha

  2. College of Plant Science, Agricultural Division, Jilin University, Changchun, 130062, China

    Ying Wang & Qing Liu

  3. College of Horticulture, Northeast Agricultural University, Harbin, 150030, China

    Yang Zhao

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  1. Yan Zhao
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  2. Ying Wang
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  3. Qing Liu
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Correspondence to Wei Sha.

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Communicated by Sergio J Ochatt.

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Zhao, Y., Wang, Y., Liu, Q. et al. Cloning of a new LEA1 gene promoter from soybean and functional analysis in transgenic tobacco. Plant Cell Tiss Organ Cult 130, 379–391 (2017). https://doi.org/10.1007/s11240-017-1234-3

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