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Agrobacterium tumefaciens-mediated transformation of Pisum sativum L. using binary and cointegrate vectors

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Abstract

Epicotyl segments and nodus expiants from etiolated seedlings of Pisum sativum were transformed using Agrobacterium tumefaciens strains GV 2260 (p35S GUS INT) and GV 3850 HPT carrying either a neomycin- or hygromycinphosphotransferase-gene as selectable markers. The transgenic character of hygromycin- or kananamycin-resistant tissue was confirmed by detection of nopaline or neomycinphosphotransferase-II- and ß-glucuronidase activity in crude extracts of resistant tissues. Up to 5 % of developing shoots from shoot proliferating nodi were regenerated via organogenesis to kanamycin-resistant plantlets. Transformation frequency in vitro was found to be influenced by expiant source, A. tumefaciens strain, pea genotype and duration of cocultivation. Acetosyringone did not increase the transformation rate.

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Abbreviations

GUS:

ß-glucuronidase

NAA:

1-naphthyl-acetic-acid

BA:

6-benzylaminopurine

NPT-II:

neomycinphosphotransferase-II

HPT:

hygromycinphosphotransferase

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Authors and Affiliations

  1. Institut für Genetik, Universität Bonn, Kirschallee 1, D-5300, Bonn 1, Federal Republic of Germany

    Andre De Kathen & Hans-Jörg Jacobsen

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  1. Andre De Kathen
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  2. Hans-Jörg Jacobsen
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Communicated by H. Lörz

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De Kathen, A., Jacobsen, HJ. Agrobacterium tumefaciens-mediated transformation of Pisum sativum L. using binary and cointegrate vectors. Plant Cell Reports 9, 276–279 (1990). https://doi.org/10.1007/BF00232301

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  • DOI: https://doi.org/10.1007/BF00232301

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