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Two tobacco AP1-like gene promoters drive highly specific, tightly regulated and unique expression patterns during floral transition, initiation and development

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Abstract

The genetic engineering of agronomic traits requires an array of highly specific and tightly regulated promoters that drive expression in floral tissues. In this study, we isolated and characterized two tobacco APETALA1-like (AP1-like) promoters (termed NtAP1La and NtAP1Lb1) in transgenic plants using the GUS reporter system, along with tissue-specific ablation analyses. Our results demonstrated that the two promoters are active in floral inflorescences but not in vegetative apical meristems or other vegetative tissues, as reflected by strong GUS staining and DT-A-mediated ablation of apical shoot tips during reproductive but not vegetative growth. We also showed that the NtAP1Lb1 promoter was more active than NtAP1La in inflorescences, as the former yielded higher frequencies and greater phenotypic evidence of tissue ablation compared to the latter. We further revealed that both promoters were uniformly expressed in the meristems of stage 1 and 2 floral buds, but were differentially expressed in floral organs later during development. While NtAP1La was found to be active in stage 4–5 carpels, later becoming confined to ovary tissue from stage 9 onwards, NtAP1Lb1 activity was apparent in all floral organs from stages 3 to 7, becoming completely absent in all floral organs from stage 11 onward. Therefore, it seems that the two tobacco promoters have acquired similar but distinct inflorescence-, floral meristem- and floral organ-specific and development-dependent regulatory features without any leaky activity in vegetative tissues. These features are novel and have rarely been observed in other flower-specific promoters characterized to date. The potential application of these promoters for engineering sterility, increasing biomass production and modifying flower architecture, as well as their putative use in flower-specific transgene excision, will be discussed.

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Abbreviations

CsVMV :

Cassava vein mosaic virus promoter

DT-A :

Diphtheria toxin A

GUS :

β-Glucuronidase

NtAP1L :

Nicotiana tabacum AP1-like

PCR:

Polymerase chain reaction

RT-PCR:

Reverse transcription polymerase chain reaction

SAM:

Shoot apical meristem

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Acknowledgments

We thank Mr. Dennis Bennett for his excellent technical assistance. This study was funded by the USDA NIFA Biotechnology Risk Assessment Research grant (2009-01067).

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Authors and Affiliations

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

    Jinjin Zhang, Guohua Yan, Zhifeng Wen & Zongrang Liu

  2. College of Life Science, Shaanxi Normal University, Xi’an, 710062, Shaanxi, China

    Jinjin Zhang

  3. The Institute of Forestry and Pomology, Beijing Academy of Agriculture and Forestry Sciences, Beijing, 100093, China

    Guohua Yan

  4. College of Horticulture, Northwest A&F University, Yangling, 712100, Shaanxi, China

    Zhifeng Wen

  5. Plant Genetics Research Unit, Agricultural Research Service, United States Department of Agriculture, Donald Danforth Plant Science Center, St Louis, MO, 63132, USA

    Young-Qiang An

  6. Department of Agricultural, Food and Nutritional Science, University of Alberta, Edmonton, AB, T6G 2P5, Canada

    Stacy D. Singer

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  1. Jinjin Zhang
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  2. Guohua Yan
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Corresponding author

Correspondence to Zongrang Liu.

Additional information

J. Zhang and G. Yan contributed equally to this work.

The nucleotide sequencing data reported here have been deposited in the GenBank database under the accession numbers KF539844 (NtAP1La), KF530845 (NtAP1Lb1) and KF530846 (NtAP1Lb2).

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Zhang, J., Yan, G., Wen, Z. et al. Two tobacco AP1-like gene promoters drive highly specific, tightly regulated and unique expression patterns during floral transition, initiation and development. Planta 239, 469–478 (2014). https://doi.org/10.1007/s00425-013-1995-9

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  • DOI: https://doi.org/10.1007/s00425-013-1995-9

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