The Agrobacterium rhizogenes pRi TL-DNA segment as a gene vector system for transformation of plants

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  • Jens Stougaard
  • Dorte Abildsten
  • ,
  • Kjeld A. Marcker, Department of Molecular Biology and Plant Physiology

A plant gene transfer system was developed from the Agrobacterium rhizogenes pRi15834 TL-DNA region. "Intermediate integration vectors" constructed from ColE1-derived plasmids served as cloning vectors in Escherichia coli and formed cointegrates into the TL-DNA after transfer to A. rhizogenes. An A. rhizogenes strain with pBR322 plasmid sequences replacing part of the TL-DNA was also constructed. Plasmids unable to replicate in Agrobacterium can integrate into this TL-DNA by homologous recombination through pBR322 sequences. No loss of pathogenicity was observed with the strains formed after integration of intermediate vectors or strains carrying pBR322 in the TL-DNA segment. Up to 15 kb of DNA have been transferred to plant cells with these systems. The T-DNA from a binary vector was cotransformed into hairy roots which developed after transfer of the wild-type pRi T-DNA. Tested on Lotus corniculatus the TL-derived vector system transformed 90% of the developed roots and the T-DNA from the binary vector was cotransformed into 60% of the roots. Minimum copy numbers of one to five were found. Both constitutive and organ-specific plant genes were faithfully expressed after transfer to the legume L. corniculatus.

TidsskriftMGG Molecular & General Genetics
Sider (fra-til)251-255
Antal sider5
StatusUdgivet - maj 1987
Eksternt udgivetJa

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