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Heterologous expression of the BABY BOOM AP2/ERF transcription factor enhances the regeneration capacity of tobacco (Nicotiana tabacum L.)

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Abstract

Gain-of-function studies have shown that ectopic expression of the BABY BOOM (BBM) AP2/ERF domain transcription factor is sufficient to induce spontaneous somatic embryogenesis in Arabidopsis (Arabidopsis thaliana (L.) Heynh) and Brassica napus (B. napus L.) seedlings. Here we examined the effect of ectopic BBM expression on the development and regenerative capacity of tobacco (Nicotiana tabacum L.) through heterologous expression of Arabidopsis and B. napus BBM genes. 35S::BBM tobacco lines exhibited a number of the phenotypes previously observed in 35S::BBM Arabidopsis and B. napus transgenics, including callus formation, leaf rumpling, and sterility, but they did not undergo spontaneous somatic embryogenesis. 35S::BBM plants with severe ectopic expression phenotypes could not be assessed for enhanced regeneration at the seedling stage due to complete male and female sterility of the primary transformants, therefore fertile BBM ectopic expression lines with strong misexpression phenotypes were generated by expressing a steroid-inducible, post-translationally controlled BBM fusion protein (BBM:GR) under the control of a 35S promoter. These lines exhibited spontaneous shoot and root formation, while somatic embryogenesis could be induced from in-vitro germinated seedling hypocotyls cultured on media supplemented with cytokinin. Together these results suggest that ectopic BBM expression in transgenic tobacco also activates cell proliferation pathways, but differences exist between Arabidopsis/B. napus and N. tabacum with respect to their competence to respond to the BBM signalling molecule.

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Abbreviations

DEX:

Dexamethasone

BAP N6 :

Benzylaminopurine

NAA:

Napthaleneacetic acid

TDZ:

Thidiazuron (N-phenyl-N′-1,2,3,-thiadiazol-5-ylurea)

TEM:

Transmission electron microscopy

SEM:

Scanning electron microscopy

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Acknowledgments

We thank Remko Offringa (University of Leiden, The Netherlands) for comments on the manuscript, Kevin Webb (USDA ARS Appalachian Fruit Research Station, Kearneysville, WV, USA) for molecular analyses, David L Bentley (Imaging Facility, The University of Arizona, Tucson, Arizona, USA) for the microscopy studies and Ahn Liseon Silverstein, Mark Demuth and Dennis Bennett (USDA ARS Appalachian Fruit Research Station) for technical assistance. E.D.J.S. was supported by fellowships from the Biotechnology Research Indonesia-Netherlands (BIORIN) research program with financial aid from the Royal Netherlands Academy of Arts and Sciences (KNAW), and the Bogor Agricultural University fellowship program, Quality for Undergraduate Education (QUE) project. H.F. was supported by a fellowship from the Science and Technology Agency of Japan.

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Authors and Affiliations

  1. United States Department of Agriculture, Agricultural Research Service, Appalachian Fruit Research Station, 2217 Wiltshire Road, Kearneysville, WV, 25430, USA

    Chinnathambi Srinivasan, Zongrang Liu & Ralph Scorza

  2. Enza Zaden Research and Development, P.O. Box 7, 1600 AA, Enkhuizen, The Netherlands

    Iris Heidmann & Joep Lambalk

  3. Plant Research International, P.O. Box 16, 6700 AA, Wageningen, The Netherlands

    Ence Darmo Jaya Supena, Ronny Joosen, Gerco Angenent, Jan B. M. Custers & Kim Boutilier

  4. Research Center for Biotechnology, Bogor Agricultural University (IPB), P.O. Box 1, Bogo, 16610, Indonesia

    Ence Darmo Jaya Supena

  5. Laboratory of Breeding Technology, National Institute of Vegetable and Tea Science (NIVTS), Ano, Mie, 514-2392, Japan

    Hiro Fukuoka

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  1. Chinnathambi Srinivasan
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Correspondence to Chinnathambi Srinivasan.

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Srinivasan, C., Liu, Z., Heidmann, I. et al. Heterologous expression of the BABY BOOM AP2/ERF transcription factor enhances the regeneration capacity of tobacco (Nicotiana tabacum L.). Planta 225, 341–351 (2007). https://doi.org/10.1007/s00425-006-0358-1

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