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Integration of Agrobacterium T-DNA into a tobacco chromosome: Possible involvement of DNA homology between T-DNA and plant DNA

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Summary

We established tobacco tumour cell lines from crown galls induced by Agrobacterium. Restriction fragments containing T-DNA/plant DNA junctions were cloned from one of the cell lines, which has a single copy of the T-DNA in a unique region of its genome. We also isolated a DNA fragment that contained the integration target site from nontransformed tobacco cells. Nucleotide sequence analyses showed that the right and left breakpoints of the T-DNA mapped ca. 7.3 kb internal to the right 25 by border and ca. 350 by internal to the left border respectively. When the nucleotide sequences around these breakpoints were compared with the sequence of the target, significant homology was seen between the region adjacent to the integration target site and both external regions of the T-DNA breakpoints. In addition, a short stretch of plant DNA in the vicinity of the integration site was deleted. This deletion seems to have been promoted by homologous recombination between short repeated sequences that were present on both sides of the deleted stretch. Minor rearrangements, which included base substitutions, insertions and deletions, also took place around the integration site in the plant DNA. These results, together with previously reported results showing that in some cases sequences homologous to those in T-DNA are present in plant DNA regions adjacent to left recombinational junctions, indicate that sequence homology between the incoming T-DNA and the plant chromosomal DNA has an important function in T-DNA integration. The homology may promote close association of both termini of a T-DNA molecule on a target sequence; then TDNA may in some cases be integrated by a mechanism at least in part analogous to homologous recombination.

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Author notes

  1. Shogo Matsumoto

    Present address: Division of Horticulture, CSIRO, GPO Box 350, 5001, Adelaide, S.A., Australia

Authors and Affiliations

  1. Department of Biology, Faculty of Science, Nagoya University, 464-01, Chikusa-ku, Nagoya, Japan

    Shogo Matsumoto, Yukihiro Ito, Tsuyoshi Hosoi, Yosuke Takahashi & Yasunori Machida

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  1. Shogo Matsumoto
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  2. Yukihiro Ito
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  3. Tsuyoshi Hosoi
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  4. Yosuke Takahashi
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  5. Yasunori Machida
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Additional information

Communicated by M. Sekiguchi

Shogo Matsumoto is on leave from Biochemical Research Institute, Nippon Menard Cosmetic Co., Ltd, Ogaki, Gifu-ken 503, Japan

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Matsumoto, S., Ito, Y., Hosoi, T. et al. Integration of Agrobacterium T-DNA into a tobacco chromosome: Possible involvement of DNA homology between T-DNA and plant DNA. Molec. Gen. Genet. 224, 309–316 (1990). https://doi.org/10.1007/BF00262423

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  • DOI: https://doi.org/10.1007/BF00262423

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