The ternary transformation system: constitutive virG on a compatible plasmid dramatically increases Agrobacterium-mediated plant transformation.
This paper describes a so-called ternary transformation system for plant cells. We demonstrate that Agrobacterium tumefaciens strain LBA4404 supplemented with a constitutive virG mutant gene (virGN54D) on a compatible plasmid is capable of very efficient T-DNA transfer to a diverse range of plant species. For the plant species Catharanthus roseus it is shown that increased T-DNA transfer results in increased stable transformation frequencies. Analysis of stably transformed C. roseus cell lines showed that, although the T-DNA transfer frequency is greatly enhanced by addition of virGN54D, only one or a few T-DNA copies are stably integrated into the plant genome. Thus, high transformation frequencies of different plant species can be achieved by introduction of a ternary plasmid carrying a constitutive virG mutant into existing A. tumefaciens strains in combination with standard binary vectors.
van der Fits
L
L
Institute of Molecular Plant Sciences, Clusius Laboratory, Leiden University, The Netherlands.
Deakin
E A
EA
Hoge
J H
JH
Memelink
J
J
D000098:Acetophenones / Q000494:pharmacology; D016960:Agrobacterium tumefaciens / Q000166:cytology / Q000187:drug effects / Q000235:genetics; D017360:Arabidopsis / Q000166:cytology / Q000235:genetics; D001426:Bacterial Proteins; D018920:Coculture Techniques; D004269:DNA, Bacterial / Q000187:drug effects / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics; D009154:Mutation; D059828:Plant Cells; D010944:Plants / Q000187:drug effects / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics; D014170:Transformation, Genetic
D000098:Acetophenones; D001426:Bacterial Proteins; D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; C036483:T-DNA; D014157:Transcription Factors; C058483:virG protein, Shigella flexneri; C051667:acetosyringone