Stable transformation of barley callus using biolistic® particle bombardment and the phosphinothricin acetyltransferase (bar) gene

Carol M. Stiff, Andrzej Kilian, Huaping Zhou, David A. Kudrna, Andris Kleinhofs

Research output: Contribution to journalArticlepeer-review

9 Scopus citations

Abstract

Suspension culture cells of barley (Hordeum vulgare L. cv. Klages) were collectec on filters and bombarded with gold particles carrying the Basta® resistance (bar) gene, or both bar and the β-glucuronidase (uidA) genes in a Biolistic® Particle Accelerator. Filters carrying the bombarded tissues were selected on basal medium with increasing concentrations (maximum of 200 mg I-1) of phosphinothricin. Surviving calli were assayed for the presence of phosphinothricin acetyltransferase activity. All positive calli were shown to have the bar gene integrated into their genome. Nine independent stable transformation events have been analyzed with substantial variation in the integration pattern and transgene copy number. Some albino plantlets were produced, but no mature plants were obtained.

Original languageEnglish (US)
Pages (from-to)243-248
Number of pages6
JournalPlant Cell, Tissue and Organ Culture
Volume40
Issue number3
DOIs
StatePublished - Mar 1995
Externally publishedYes

Keywords

  • Hordeum vulgare L.
  • accelerated particle
  • integration
  • methylation
  • suspension culture

ASJC Scopus subject areas

  • Horticulture

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