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Regeneration and transformation in adult plants of Campanula species

  • Genetic Transformation and Hybridization
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Abstract

Adult plants are known for recalcitrance when it comes to adventitious organ formation and regeneration. Methods used for regeneration in explants from seedlings of Campanula carpatica failed to work for explants from adult plants of the same species. The present investigation generated efficient regeneration methods for mature specimens of four species of Campanula, C. carpatica, C. haylodgensis, C. portenschlagiana and C. poscharskyana. Petiole explants from dark-grown in vitro shoot cultures grown from nodal cuttings of adult plants regenerated successfully (95%), while explants from light-grown in vitro shoot cultures and greenhouse-grown plants regenerated at 12% and zero percentage, respectively. Dark-treatment, along with media manipulation with plant growth regulators, further enhanced regenerative capacity of the explants. A MS-based medium containing 10mg l −1 TDZ and 0.25 mg l−1 NAA was the most efficient regeneration medium. Transgenic shoots from C. carpatica (3%) and C. haylodgensis (1%) and transgenic callus from all species were produced using Agrobacterium tumefaciens, and transformation was confirmed by histochemical and Southern blot analyses. Protocols developed in this study may be useful for achieving efficient regeneration and transformation of recalcitrant adult plants.

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Abbreviations

BA:

6-Benzylaminopurine

NAA:

α-Naphthalene acetic acid

TDZ:

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

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Acknowledgments

The project was funded by the Danish Ministry of Agriculture (93S-2466-Å97-01023), the Danish Campanula Growers: Gartneriet Thoruplund A/S (Odense) and Gartneriet Elmegaard ApS (Skælskør) (SS and MS), and the University of Hannover (MS and HM). The authors thank Professor Gerco Angenent, Plant Research International, The Netherlands, for providing the bacterial strains and Dr. Bjarne Stummann for critical reading of the manuscript. We acknowledge Prof. Errol Hewett, Massey University, New Zealand for reading the manuscript for clarity and language. We also thank Thanussa Thuraisingam and Annette Steding for their excellent technical assistance.

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Authors and Affiliations

  1. Department of Plant Biology and Forest Genetics, Swedish University of Agricultural Sciences, P.O. Box 7080, 750 07, Uppsala, Sweden

    Sridevy Sriskandarajah

  2. Faculty of Natural Sciences, Institute for Ornamental and Woody Plant Science, University of Hannover, Herrenhaeuser Str. 2, 30419, Hannover, Germany

    Heiko Mibus & Margrethe Serek

Authors
  1. Sridevy Sriskandarajah
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  2. Heiko Mibus
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  3. Margrethe Serek
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Correspondence to Sridevy Sriskandarajah.

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Communicated by P. Kumar.

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Sriskandarajah, S., Mibus, H. & Serek, M. Regeneration and transformation in adult plants of Campanula species. Plant Cell Rep 27, 1713–1720 (2008). https://doi.org/10.1007/s00299-008-0590-1

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  • DOI: https://doi.org/10.1007/s00299-008-0590-1

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