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A Modular Vector for Agrobacterium Mediated Transformation of Wheat

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An Erratum to this article was published on 01 June 2000

Abstract

Wheat (cv Chinese Spring) tissues were transformed using Agrobacterium tumefasciens and a new plasmid modular vector, pMVTBP. We constructed pMVTBP with unique restriction sites connecting (1) the CaMV 35S promoter, (2) a Kozak sequence, (3) the FLAG epitope, (4) the (His)6 epitope, (5) a coding region (for wheat TATA Binding Protein, wTBP) and (6) the CaMV 35S 3′UTR. This vector thus allows easy exchange of different regulatory or coding sequences. Explants of either germinating mature seeds, or immature embryos, were induced to callus for up to two weeks, treated with virulence-induced bacteria for one hour, then regenerated into plantlets. Transient expression of a GUS reporter gene, assayed at about one week, occurred in 10–12% of calluses. Expression of the FLAG-tagged wTBP was also detected, by immunostaining. Stable expression, by selective growth on geneticin, and by GUS expression at about six weeks, occurred in 1–2% of calluses, quite comparable to that achieved by other methods.

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Authors and Affiliations

  1. Institute for Chemistry and Cell Biology, Harvard Medical School, 240 Longwood Ave., Boston, MA, 02115, USA

    Noël R. Peters

  2. Biology Department, University of Massachusetts, Boston

    Steven Ackerman & Elizabeth A. Davis

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  1. Noël R. Peters
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  2. Steven Ackerman
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  3. Elizabeth A. Davis
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Additional information

An erratum to this article is available at http://dx.doi.org/10.1007/BF02824027.

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Peters, N.R., Ackerman, S. & Davis, E.A. A Modular Vector for Agrobacterium Mediated Transformation of Wheat. Plant Molecular Biology Reporter 17, 323–331 (1999). https://doi.org/10.1023/A:1007686408369

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