Plant Molecular Biology

, Volume 34, Issue 6, pp 913–922 | Cite as

Frequent collinear long transfer of DNA inclusive of the whole binary vector during Agrobacterium-mediated transformation

  • Allan Wenck
  • Mihály Czakó
  • Ivan Kanevski
  • László Márton

Abstract

When Agrobacterium was used to transform Nicotiana plumbaginifolia protoplasts and Arabidopsis thaliana roots and seedlings, a large number of plants were found in which not only the T-region defined by the border repeat sequences but the entire binary vector was integrated, as determined by both PCR and Southern analysis techniques. N. plumbaginifolia protoplast co-cultivation experiments yielded 3 out of 5 transformants with collinear sequence past the left border. In Arabidopsis root transformation experiments, 33% (6/18) of the transformants had T-DNA which exceeded the left border repeat. Vacuum infiltration of Arabidopsis seedlings produced even a greater percentage of transformants with sequences outside the left border repeat (62%, 39/63). The long transfer DNA cosegregated with the T-region encoded hygromycin resistance in the T2 progeny eliminating the possibility that long transfer DNA was of extrachromosomal or Agrobacterium origin. The high frequency of long transfer after vacuum infiltration of A. thaliana needs to be considered when analyzing T-DNA tagged mutants.

Agrobacterium tumefaciens Arabidopsis thaliana Nicotiana plumbaginifolia root transformation T-DNA transfer vacuum infiltration 

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Copyright information

© Kluwer Academic Publishers 1997

Authors and Affiliations

  • Allan Wenck
    • 1
  • Mihály Czakó
    • 1
  • Ivan Kanevski
    • 1
  • László Márton
    • 1
  1. 1.Department of BiologyUniversity of South CarolinaColumbiaUSA
  2. 2.Forest Biotechnology Group, Dept. of ForestryNorth Carolina State UniversityRaleighUSA
  3. 3.Biomedical ConsultingPiscatawayUSA

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