High-efficiency gene transfer to recalcitrant plants by Agrobacterium tumefaciens

J. Ke, R. Khan, T. Johnson, D. A. Somers, A. Das

Research output: Contribution to journalArticlepeer-review

28 Scopus citations

Abstract

Agrobacterium tumefaciens efficiently transforms most plants. A few dicotyledonous plants and most monocotyledonous plants are, however, recalcitrant to A. tumefaciens infection. We investigated whether the constitutive synthesis of a high level of the T-strand DNA intermediate can improve the transformation efficiency of plants. We previously described a mutation in the vir gene regulator virG, virGN54D, that allows constitutive expression of the vir genes. We also described the isolation of a mutant plasmid that is present at a significantly high level in A. tumefaciens. The two mutations were combined to produce an A. tumefaciens strain that synthesizes a high level of T-strand DNA in an inducer-independent manner. DNA transfer efficiency of the mutant was measured by monitoring β-glucuronidase (GUS) expression in a transient transfer assay. A significant increase in the efficiency of DNA transfer to both rice and soybean was observed with the double mutant. The presence of virGN54D had a major positive effect on transformation efficiency.

Original languageEnglish (US)
Pages (from-to)150-156
Number of pages7
JournalPlant Cell Reports
Volume20
Issue number2
DOIs
StatePublished - 2001

Bibliographical note

Funding Information:
Acknowledgements This work was supported by USDA prime grant no. 96–34340–2711 from Purdue University.

Keywords

  • Agrobacterium tumefaciens
  • GUS gene expression
  • Plant transformation
  • Rice
  • Soybean

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