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Stable transformation of barley tissue culture by particle bombardment

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Summary

Suspension culture cells of barley (Hordeum vulgare L. cv Pokko) were stably transformed with two separate plasmids containing genes coding for neomycin phosphotransferase II and β-glucuronidase, respectively. Transformed cultures were selected with the antibiotic GeneticinR. Enzymatic activity was tested in the GeneticinR resistant cultures, and in 96% of them neomycin phosphotransferase could be detected. The non-selected marker, detected as β-glucuronidase activity, was expressed in 40% of the resistant cultures. Stable transformation was confirmed with Southern blot hybridization.

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Abbreviations

GUS:

β-glucuronidase of E. coli

uidA :

gene coding for

GUS:

G418

GeneticinR :

O-2-amino-2,7-dideoxy-D-glycero-α-D-glucoheptopyranosyl[1-4]-O-3-deoxy-4C-methyl-3-[methylamino]-β-L-arabinopyranosyl-D-streptamine disulfate salt

NPTII:

neomycin phosphotransferase II of Tn5

nptII :

gene coding for NPTII

2,4-D:

2,4-dichlorophenoxyacetic acid

X-Gluc:

5-bromo-4-chloro-3-indolylβ-D-glucuronic acid cyclohexylammonium salt

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Author notes

  1. J. Mendez Lozano

    Present address: Department of Plant Genetic Engineering, CINVESTAV, Irapuato, Mexico

Authors and Affiliations

  1. VTT, Biotechnical Laboratory, PL 202, SF-02151, Espoo, Finland

    A. Ritala, L. Mannonen, M. Salmenkallio-Marttila, U. Kurtén, R. Hannus & V. Kauppinen

  2. Institute of Biotechnology, University of Helsinki, SF-00380, Helsinki, Finland

    K. Aspegren, J. Mendez Lozano & T. H. Teeri

Authors
  1. A. Ritala
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  2. L. Mannonen
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  3. K. Aspegren
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  4. M. Salmenkallio-Marttila
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  5. U. Kurtén
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  6. R. Hannus
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  7. J. Mendez Lozano
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  8. T. H. Teeri
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  9. V. Kauppinen
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Additional information

Communicated by H. Lörz

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Ritala, A., Mannonen, L., Aspegren, K. et al. Stable transformation of barley tissue culture by particle bombardment. Plant Cell Reports 12, 435–440 (1993). https://doi.org/10.1007/BF00234708

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  • DOI: https://doi.org/10.1007/BF00234708

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