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Creation and analysis of a novel chimeric promoter for the complete containment of pollen- and seed-mediated gene flow

  • Genetic Transformation and Hybridization
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Abstract

Effective containment of gene flow in transgenic plants requires a promoter that is highly specific for male and female gametes or tissues. Here, we report the creation of a novel pollen-, stigma- and carpel-specific (PSC) promoter through the fusion of the pollen-specific LAT52 and carpel-specific AGL5 enhancers to a stigma-specific SLG promoter. Gene expression analysis showed that fusion of the LAT52 enhancer to the SLG promoter enables the latter to gain pollen-specific activity while the acquirement of carpel-specific activity requires the correct orientation of the inserted AGL5 enhancer in the PSC promoter, and only a forward- but not a reverse-oriented one is functional. The resulting fPSC promoter, when fused to DT-A, generated at least three aberrant gynoecium phenotypes. Type I plants exhibited shortened stigmatic tissues, resembling plants containing the DT-A gene controlled by the SLG promoter. However, type II and III plants displayed partial or complete ablation of gynoecia, and were unable to support the reproductive process. Type II and III plants also produced severely perturbed anthers and pollen in comparison to type I or SLG::DT-A plants, and transgenic pollen grains were unable, when out-crossed with control plants, to pass the transgene to the next generation in all plants examined, indicating that they are selectively eliminated. This tissue-specific ablation or perturbation is highly specific, and does not compromise vegetative growth. Evidently, the fPSC promoter faithfully acquires tissue specificity from the incorporated enhancers and promoter, and should have a practical application for transgene containment in non-fruit and -grain producing plant crops.

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Acknowledgments

We thank Dennis Bennett and Lin Zhang for the help with generating transgenic plants, Dr. June Nasrallah for providing SLG13 promoter and Dr. Stacy Singer for critical reading of the manuscript. This work was supported by a grant from US Department of Agriculture (2006-03701) to ZL.

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Authors and Affiliations

  1. USDA-ARS, Appalachian Fruit Research Station, 2217 Wiltshire Road, Kearneysville, WV, 25430, USA

    Zongrang Liu

  2. Department of Biology, West Virginia University, Morgantown, WV, 26506-6057, USA

    Changhe Zhou

  3. Institute of Plant Biology, College of Life Science, National Taiwan University, Taipei, 106, Taiwan

    Keqiang Wu

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  1. Zongrang Liu
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  2. Changhe Zhou
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Correspondence to Zongrang Liu or Keqiang Wu.

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Communicated by R. Schmidt.

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Liu, Z., Zhou, C. & Wu, K. Creation and analysis of a novel chimeric promoter for the complete containment of pollen- and seed-mediated gene flow. Plant Cell Rep 27, 995–1004 (2008). https://doi.org/10.1007/s00299-008-0522-0

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