Development of auxotrophic Agrobacterium tumefaciens for gene transfer in plant tissue culture

Jason I. Collens, Dae R. Lee, Amy M. Seeman, Wayne R. Curtis

Research output: Contribution to journalArticlepeer-review

17 Scopus citations

Abstract

Auxotrophic strains of Agrobacterium tumefaciens were generated for use in liquid co-culture with plant tissue for transient gene expression. Twenty-one auxotrophs were recovered from 1,900 tetracycline-resistant insertional mutants generated with a suicide vector transposon mutagenesis system. Twelve of these auxotrophs were characterized on a nutrient matrix. Isolates were screened for growth in plant cell and root culture, and three auxotrophs were identified that had limited growth: adenine (ade-24), leucine (leu-27), and cysteine (cys-32). Ade-24 displayed poor T-DNA delivery in a transient expression test delivering GUS from a binary vector, while cys-32 displayed the best ability to deliver DNA of these three auxotrophs. The growth yield of cys-32 on cysteine was assessed to provide a quantitative basis for co-culture nutrient supplementation. The utility of cys-32 for delivering T-DNA to plant tissues is demonstrated, where an 85-fold enhancement in GUS expression over wild-type A. tumefaciens was achieved.

Original languageEnglish (US)
Pages (from-to)890-896
Number of pages7
JournalBiotechnology Progress
Volume20
Issue number3
DOIs
StatePublished - May 1 2004

All Science Journal Classification (ASJC) codes

  • Biotechnology

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