Genetic analysis of transfer and stabilization of <i>Agrobacterium</i>
 DNA in plant cells

Joos, Henk; Timmerman, Benedikt; Montagu, Marc Van; Schell, Jeff

doi:10.1002/j.1460-2075.1983.tb01716.x

Cited by 77 publications

(42 citation statements)

References 36 publications

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“…In contrast mutants that lack the left border repeat can still efficiently transfer the onc-genes to the plant cell (13,21).…”

Section: Discussionmentioning

confidence: 99%

“…Studies on the border fragments of the T-region showed that a deletion of the left border repeat did not result in any detectable effect on the T-region transfer process (13,21), while deletion of the right border of the nopaline T-region (35) and of the octopine T-region (30) resulted in the loss of oncogenicity. The onc-genes and the vir-genes essential for tumor induction were still intact in these deletion mutants.…”

Section: N a Isolation Proceduresmentioning

confidence: 99%

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Functional analysis of the Agrobacterium tumefaciens octopine Ti-plasmid left and right T-region border fragments

et al. 1987

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SummaryBorder fragments of the octopine Ti-plasmid were tested for their ability to restore tumorigenicity of an avirulent mutant carrying a deleted right border. It was found that neither introduction of left border fragments nor that of small right border fragments at the position of the deletion resulted in a complete restoration of oncogenicity. However, insertion of a larger right border fragment in the deletion mutant gave fully oncogenic strains. In the latter case sequences to the right side of the right border repeat were found to be responsible for a complete restoration of oncogenicity. Also a left border repeat inserted together with this enhancer sequence fully restored the oncogenicity of the deletion mutant. The enhancer-sequence on itself was not able to mediate the transfer of the T-region to the plant cell. Border fragments inserted in inverted orientation in the deletion mutant were able to mediate the transfer of the T-region to the plant cell, but at a reduced frequency.

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“…In contrast mutants that lack the left border repeat can still efficiently transfer the onc-genes to the plant cell (13,21).…”

Section: Discussionmentioning

confidence: 99%

Section: N a Isolation Proceduresmentioning

confidence: 99%

Functional analysis of the Agrobacterium tumefaciens octopine Ti-plasmid left and right T-region border fragments

et al. 1987

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“…Because T-DNA transfer is apparently a polar process, initiating at the right border and continuing to (and perhaps beyond) the left border (Caplan et al, 1985;Joos et al, 1983;Miranda et al, 1992;Peralta et al, 1986), we expected that plants generated using pBSG-t w o u l d have a higher probability of receiving and integrating the mas2'-gusA gene than would plants generated using pBSG-2. Our data, however, indicated that there was no major difference in the transfer of the gusA gene whether this gene were located close to the left or to the right T-DNA border.…”

Section: Identification Of T-dna and Binary Vector 'Backbone' Single-mentioning

confidence: 99%

Integration of T‐DNA binary vector ‘backbone’ sequences into the tobacco genome: evidence for multiple complex patterns of integration

Kononov

Bassüner

Gelvin³

1997

The Plant Journal

220

139

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“…Deletion of the right-hand border repeat abolishes tumorigenesis (23,32,37,43,55); in contrast, removal of the left border does not detectably affect virulence (31,38). Inversion of the rightborder repeat in the Ti plasmid severely attenuates tumorigenesis, indicating that orientation affects its activity (37,55).…”

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confidence: 99%

Agrobacterium tumefaciens transfers extremely long T-DNAs by a unidirectional mechanism

et al. 1992

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During crown gall tumorigenesis, part of the Agrobacterium tumefaciens tumor-inducing (Ti) plasmid, the T-DNA, integrates into plant DNA. Direct repeats define the left and right ends of the T-DNA, but tumorigenesis requires only the right-hand repeat. Virulence (vir) genes act in trans to mobilize the T-DNA into plant cells. Transfer of T-DNA begins when the VirD endonuclease cleaves within the right-hand border repeat. Although the T-DNA right-border repeat promotes T-DNA transmission best in its normal orientation, an inverted right border exhibits reduced but significant activity. Two models may account for this diminished tumorigenesis. The right border may function bidirectionally, with strong activity only in its wild-type orientation, or it may promote T-DNA transfer in a unidirectional manner such that, with an inverted right border, transfer proceeds around the entire Ti plasmid before reaching the T-DNA. To determine whether a substantial portion of the Ti plasmid is transferred to plant cells, as predicted by the unidirectional-transfer hypothesis, we examined T-DNAs in tumors induced by strains containing a Ti plasmid with a right border inverted with respect to the T-DNA oncogenes. These tumors contained extremely long T-DNAs corresponding to most or all of the Ti plasmid. To test whether the right border can function bidirectionally, we inserted T-DNAs with either a properly oriented or an inverted right border into a specific site in the A. tumefaciens chromosome. A border situated to transfer the oncogenes first directed T-DNA transfer even from the bacterial chromosome, whereas a border in the opposite (inverted) orientation did not transfer the oncogenes to plant cells. Our results indicate that the right-border repeat functions in a unidirectional manner.

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Genetic analysis of transfer and stabilization of Agrobacterium DNA in plant cells

Cited by 77 publications

References 36 publications

Functional analysis of the Agrobacterium tumefaciens octopine Ti-plasmid left and right T-region border fragments

Functional analysis of the Agrobacterium tumefaciens octopine Ti-plasmid left and right T-region border fragments

Integration of T‐DNA binary vector ‘backbone’ sequences into the tobacco genome: evidence for multiple complex patterns of integration

Agrobacterium tumefaciens transfers extremely long T-DNAs by a unidirectional mechanism

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