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Comparison of the MpEF1α and CaMV35 promoters for application in Marchantia polymorpha overexpression studies

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Abstract

Constitutive promoters are essential tools for analyses of gene functions by transgenic approaches. For overexpression and silencing studies of genes, a ubiquitous and strong expression of genes under investigation as well as selection markers is preferred. For future applications in the emerging basal plant model system Marchantia polymorpha, a liverwort, activities of the viral 35S cauliflower mosaic virus promoter and the endogenous elongation factor 1α (MpEF1α) promoter were analyzed. Expression of the reporter gene β-glucuronidase (GUS), driven by the CaMV35 and MpEF1α promoters, was compared throughout plant development. Significant differences were observed between the two promoter activities. The CaMV35 promoter yields a weak reporter gene expression in the meristematic zones but drives a strong expression in the thallus. The MpEF1α promoter causes a strong meristematic GUS expression and is more active in female sexual tissues. Overall, the MpEF1α promoter seems to be the better option for obtaining a strong and ubiquitous transgene expression. Furthermore, a whole mount in situ hybridization protocol for Marchantia was established. Analysis of MpEF1α mRNA transcript in intact, whole tissues showed an expression pattern that is overall similar to the pattern of the GUS reporter gene expression driven by the MpEF1α promoter, including strong expression in meristematic zones. The whole mount technique reported here can be used to determine the mRNA expression in intact gemmae and archegonia, and has the potential to be applied for screening large numbers of transgenic plants, for instance to identify knock-down mutants.

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Acknowledgments

The pGWB2 and pGWB3 vectors (Nakagawa et al. 2007) were a kind gift of Tsuyoshi Nakagawa, Research Institute of Molecular Genetics, Shimane University, Japan. This work was supported by the Deutsche Forschungsgemeinschaft (ZA259/4 to S.Z.). We are grateful for ongoing support from the Botanical Garden of Osnabrück in cultivating Marchantia.

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Author notes

  1. Oliver Zobell

    Present address: European Commission, Joint Research Centre, Institute for Reference Materials and Measurements, Brussels, Belgium

Authors and Affiliations

  1. Department of Botany, University of Osnabrück, Barbarastraße 11, 49069, Osnabrück, Germany

    Felix Althoff, Sarah Kopischke, Oliver Zobell & Sabine Zachgo

  2. Graduate School of Biostudies, Kyoto University, Kyoto, 606-8502, Japan

    Kentaro Ide, Kimitsune Ishizaki & Takayuki Kohchi

Authors
  1. Felix Althoff
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  2. Sarah Kopischke
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  3. Oliver Zobell
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  5. Kimitsune Ishizaki
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  6. Takayuki Kohchi
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  7. Sabine Zachgo
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Correspondence to Sabine Zachgo.

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Felix Althoff, Sarah Kopischke, and Oliver Zobell have contributed equally to this work.

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Althoff, F., Kopischke, S., Zobell, O. et al. Comparison of the MpEF1α and CaMV35 promoters for application in Marchantia polymorpha overexpression studies. Transgenic Res 23, 235–244 (2014). https://doi.org/10.1007/s11248-013-9746-z

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  • DOI: https://doi.org/10.1007/s11248-013-9746-z

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