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Agrobacterium tumefaciens-mediated transformation of Campanula carpatica: factors affecting transformation and regeneration of transgenic shoots

  • Genetic Transformation and Hybridization
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Abstract

An efficient transformation system for Campanula carpatica was developed using Agrobacterium tumefaciens strains LBA4404 (harbouring the plasmid pBI121), and AGL0 (harbouring the plasmid pBEO210). This is the first report on the transformation of C. carpatica. Various factors affecting the transformation efficiency and subsequent regeneration were identified. The age of seedlings from which the explants for transformation studies were taken, and the growth conditions under which the seedlings were grown had a significant influence on the production of transformed shoots. Hypocotyls taken from 12-day-old seedlings grown in the dark were the most productive, with up to 25% of hypocotyls producing transformed shoots. Explants taken from 5-week-old seedlings produced only transformed callus. The medium used for co-cultivation and incubation also had a significant influence on transformation frequency and shoot regeneration. The cultivar “Blue Uniform” was more responsive than “White Uniform”. Both bacterial strains and plasmids were equally effective in producing transformed tissue. Transformed shoots were selected on kanamycin medium, and the presence of the uidA and nptII genes in those selected shoots was confirmed by β-glucuronidase and ELISA analyses, respectively.

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Abbreviations

BAP:

6-Benzylaminopurine

NAA:

α-Naphthalene acetic acid

TDZ:

Thidiazuron

BU:

Blue Uniform

WU:

White Uniform

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Acknowledgements

The project was funded by the Danish Ministry of Agriculture (93S-2466-Å97-01023) and Danish Campanula Growers: Gartneriet Thoruplund A/S (Odense), Gartneriet PKM ApS (Odense) and Gartneriet Elmegaard ApS (Skælskør). The authors would like to thank the growers Jørgen Andersen, Kristian Madsen and Per Elmegård for their support and cooperation.

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Authors and Affiliations

  1. Department of Agricultural Sciences, Royal Veterinary and Agricultural University, 40 Thorvaldsensvej, 1871, Frederiksberg C, Copenhagen, Denmark

    Sridevy Sriskandarajah, Stefan Frello & Margrethe Serek

  2. Department of Plant Biology, Plant Biochemistry Laboratory, Royal Veterinary and Agricultural University, 40 Thorvaldsensvej, 1871, Frederiksberg C, Copenhagen, Denmark

    Kirsten Jørgensen

  3. Department of Horticulture, Institute for Floriculture, Tree Nursery Science and Plant Breeding, University of Hannover, Herrenhaeuser Strasse 2, 30419, Hannover, Germany

    Margrethe Serek

Authors
  1. Sridevy Sriskandarajah
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  2. Stefan Frello
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  3. Kirsten Jørgensen
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  4. Margrethe Serek
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Corresponding author

Correspondence to Sridevy Sriskandarajah.

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Communicated by H. Lörz

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Sriskandarajah, S., Frello, S., Jørgensen, K. et al. Agrobacterium tumefaciens-mediated transformation of Campanula carpatica: factors affecting transformation and regeneration of transgenic shoots. Plant Cell Rep 23, 59–63 (2004). https://doi.org/10.1007/s00299-004-0797-8

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  • DOI: https://doi.org/10.1007/s00299-004-0797-8

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