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Activity of the 5′ regulatory regions of the rice polyubiquitin rubi3 gene in transgenic rice plants as analyzed by both GUS and GFP reporter genes

  • Genetic Transformation and Hybridization
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Abstract

Ubiquitin is an abundant protein involved in protein degradation and cell cycle control in plants and rubi3 is a polyubiquitin gene isolated from rice (Oryza sativa L.). Using both GFP and GUS as reporter genes, we analyzed the expression pattern of the rubi3 promoter as well as the effects of the rubi3 5′-UTR (5′ untranslated region) intron and the 5′ terminal 27 bp of the rubi3 coding sequence on the activity of the promoter in transgenic rice plants. The rubi3 promoter with the 5′-UTR intron was active in all the tissue and cell types examined and supported more constitutive expression of reporter genes than the maize Ubi-1 promoter. The rubi3 5′-UTR intron mediated enhancement on the activity of its promoter in a tissue-specific manner but did not alter its overall expression pattern. The enhancement was particularly intense in roots, pollen grains, inner tissue of ovaries, and embryos and aleurone layers in maturing seeds. The translational fusion of the first 27 bp of the rubi3 coding sequence to GUS gene further enhanced GUS expression directed by the rubi3 promoter in all the tissues examined. The rubi3 promoter should be an important addition to the arsenal of strong and constitutive promoters for monocot transformation and biotechnology.

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Abbreviations

5′-UTR:

5′ Untranslated region

CaMV:

Cauliflower mosaic virus

DIC:

Differential interference contrast

GFP:

Green fluorescent protein

GUS:

β-Glucuronidase

IME:

Intron-mediated enhancement of gene expression

MU:

4-Methylumbelliferone

MUG:

4-Methylumbelliferyl-β-d-glucoside

NOS:

Nopaline synthase

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Acknowledgments

We thank Drs. N. S. Allen, E. Johannes, and J. Xu for their technical assistance with the imaging work at CMIF, NCSU, the Plant Analysis Group at Syngenta Biotechnology, Inc., for assaying the transgene copy number using realtime PCR, Dr. K. Azhakanandam for technical assistance in rice transformation, as well as Drs. J. Thomas, J. Shurtleff, and C. Saravitz, and J. Edwards for assistance in using the North Carolina State University Phytotron facility for plant maintenance. We also wish to thank Dr. T. Sit for providing the plasmid pRTL2-sGFP and Dr. P. Samadder for the plasmid pPSRG30. We are grateful to Drs. G. Allen, N. S. Allen, R. Dewey, and J. Nicholson for helpful discussions on the project. This work was supported in part by grants from the Center for Turfgrass Environmental Research and Education, North Carolina State University, The Consortium for Plant Biotechnology Research, Inc., and Syngenta Biotechnology, Inc.

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  1. Jianli Lu

    Present address: Department of Plant Biology, North Carolina State University, Raleigh, NC, 27695, USA

Authors and Affiliations

  1. Department of Crop Science, North Carolina State University, Raleigh, NC, 27695, USA

    Jianli Lu, Elumalai Sivamani & Rongda Qu

  2. Syngenta Biotechnology, Inc., Research Triangle Park, NC, 27709, USA

    Xianggan Li

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  1. Jianli Lu
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  2. Elumalai Sivamani
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Correspondence to Rongda Qu.

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Communicated by P. Ozias-Akins.

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Lu, J., Sivamani, E., Li, X. et al. Activity of the 5′ regulatory regions of the rice polyubiquitin rubi3 gene in transgenic rice plants as analyzed by both GUS and GFP reporter genes. Plant Cell Rep 27, 1587–1600 (2008). https://doi.org/10.1007/s00299-008-0577-y

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