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Regeneration and Agrobacterium-mediated transformation of hop (Humulus lupulus L.)

  • Genetic Transformation and Hybridization
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Abstract

An efficient procedure for direct organogenesis and regeneration of hop (Humulus lupulus L.) was established. For the first time Agrobacterium-mediated genetic transformation of hop (cv. "Tettnanger") was achieved. Shoot internodes from in vitro cultures were identified as the most suitable type of explant for regeneration. Using this type of explant, a shoot-inducing medium was developed that supported direct organogenesis of approximately 50% of the explants. Plantlets were successfully rooted and transferred to the greenhouse. Overall, in less than 6 months hop cultures propagated in vitro were regenerated to plants in the greenhouse. Agrobacterium-mediated genetic transformation was performed with the reporter gene GUS (β-glucuronidase). The presence and function of transgenes in plants growing in the greenhouse was verified by PCR (polymerase chain reaction) and enzyme assay for GUS activity, respectively. We have obtained 21 transgenic plants from 1,440 explants initially transformed, yielding an overall transformation efficiency of 1.5%.

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Fig. 1.
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Abbreviations

BAP :

6-Benzylaminopurine

GA 3 :

Gibberellic acid

GUS :

β-Glucuronidase

IAA :

Indole-3-acetic acid

IBA :

Indole-3-butyric acid

NAA :

α-Naphthaleneacetic acid

nptII :

Neomycin phosphotransferase II

PCR :

Polymerase chain reaction

TDZ :

1-Phenyl-3-(1,2,3-thiadiazol-5-yl) urea (thidiazuron)

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Acknowledgements

This work was funded by "Ministerium Ländlicher Raum, Baden-Württemberg" and "Hopfenpflanzerverband Tettnang e.V.", whose support is gratefully acknowledged. Also we would like to thank Ute Born and Monika Stanke for their excellent technical assistance.

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Authors and Affiliations

  1. Department of Plant Breeding and Biotechnology, Institute of Plant Breeding, Seed Science, and Population Genetics, University of Hohenheim, 70599, Stuttgart, Germany

    C. Horlemann, A. Schwekendiek, M. Höhnle & G. Weber

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  1. C. Horlemann
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  2. A. Schwekendiek
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  3. M. Höhnle
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  4. G. Weber
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Correspondence to G. Weber.

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Communicated by H. Lörz

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Horlemann, C., Schwekendiek, A., Höhnle, M. et al. Regeneration and Agrobacterium-mediated transformation of hop (Humulus lupulus L.). Plant Cell Rep 22, 210–217 (2003). https://doi.org/10.1007/s00299-003-0676-8

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  • DOI: https://doi.org/10.1007/s00299-003-0676-8

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