DNA rearrangement associated with the integration of T-DNA in tobacco: an example for multiple duplications of DNA around the integration target

Authors

  • Toshiharu Ohba,

    1. Department of Biology, School of Science, Nagoya University, Chikusa-ku, Nagoya 464-01, Japan
    Search for more papers by this author
    • Present address: Bioresearch Center, Takara Shuzo Co. Ltd, 3-4-1 Seta, Ohtsu, Shiga-ken 520-21, Japan.

  • Yasushi Yoshioka,

    1. Department of Biology, School of Science, Nagoya University, Chikusa-ku, Nagoya 464-01, Japan
    Search for more papers by this author
  • Chiyoko Machida,

    1. Cell and Information Group, PRESTO, Research Development Corporation of Japan, Aichi Institute of Technology, Yakusa, Toyota 470-03, Japan
    Search for more papers by this author
  • Yasunori Machida

    Corresponding author
    1. Department of Biology, School of Science, Nagoya University, Chikusa-ku, Nagoya 464-01, Japan
    Search for more papers by this author

*For correspondence (fax +81 52 789 2966).

Abstract

Transferred DNA (T-DNA) of the tumor-inducing (Ti) plasmid is transferred from Agrobacterium tumefaciens to plant cells and is stably integrated into the plant nuclear genome. By the inverse polymerase chain reaction DNA fragments were amplified that contained the T-DNA/plant DNA junctions from the total DNA of a transgenic tobacco plant that had a single copy of the T-DNA in a repetitive region of its genome. A DNA fragment containing the target site was amplified from the total DNA of non-transformed tobacco by the polymerase chain reaction using high-stringency conditions. Comparison of the nucleotide sequence of the target site with those of the T-DNA/plant DNA junctions revealed that various duplications of short stretches of nucleotide sequences around the target and in the incoming T-DNA had accompanied the integration of the T-DNA. A deletion of 16 bp at the target site was also found and the target site was similar, in terms of nucleotide sequence, to regions around the breakpoints of the T-DNA. This finding provides a clear example of the occurrence of complex rearrangements during the integration of T-DNA.

Ancillary