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The MsPRP2 promoter enables strong heterologous gene expression in a root-specific manner and is enhanced by overexpression of Alfin 1

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Abstract

Promoter specificity and efficiency of utilization are essential for endogenous and transgene expression. Selective root expression remains to be defined in terms of both promoter elements and transcription factors that provide high levels of ubiquitous expression. We characterized expression from the MsPRP2 promoter with the green fluorescent protein (GFP) reporter transgene in alfalfa (Medicago sativa) and found that a promoter fragment (+1 to −652 bp) retained the root and callus specificity of the endogenous MsPRP2 gene and hence this promoter fragment contains elements necessary for root-specific expression. The strong ubiquitous expression obtained from this promoter was comparable to that of the CaMV 35S promoter in roots and was enhanced by transgenic overexpression of Alfin 1, a root- and callus-specific transcription factor in alfalfa. No transgenic expression was obtained in leaves with this promoter in the presence or absence of Alfin 1. The increased expression of GFP in alfalfa containing the Alfin 1 transgene confirms the function of Alfin 1 binding sites in the MsPRP2 promoter fragment and also indicates that Alfin 1 concentrations are limiting for maximal expression in calli and roots. These findings characterize the MsPRP2 promoter as a novel root- and callus-specific promoter of plant origin that can be used as an effective tool for strong root-directed gene expression. In addition, we have demonstrated that the signal sequence of MsPRP2 can be used for efficient secretion of transgene products from callus and roots.

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Abbreviations

CaMV :

Cauliflower mosaic virus

GFP :

Green fluorescent protein

MsPRP2 :

Medicago sativa proline rich protein 2

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Acknowledgements

We would like to thank Dr. Jim Haseloff for his gift of the msgfp5 plasmid. Our thanks also to Drs. R. Trelease and R. Flynn for discussions. This work was supported in part by a grant from the Wallace Research Foundation.

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Author notes

  1. Babu Valliyodan

    Present address: Department of Agronomy, Plant Sciences Unit, University of Missouri, Columbia, MO 65211, USA

Authors and Affiliations

  1. Department of Plant Biology, Arizona State University, Main Campus PO Box 871601, Tempe, AZ 85287–01601, USA

    Ilga Winicov, Babu Valliyodan, Lingru Xue & J. Kenneth Hoober

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  1. Ilga Winicov
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  2. Babu Valliyodan
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Correspondence to Ilga Winicov.

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Winicov, I., Valliyodan, B., Xue, L. et al. The MsPRP2 promoter enables strong heterologous gene expression in a root-specific manner and is enhanced by overexpression of Alfin 1 . Planta 219, 925–935 (2004). https://doi.org/10.1007/s00425-004-1296-4

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