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

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 PC...

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Bibliographic Details
Published in:Plant molecular biology 1997-08, Vol.34 (6), p.913-922
Main Authors: Wenck, A, Czako, M, Kanevski, I, Marton, L
Format: Article
Language:English
Subjects:
Agrobacterium tumefaciens
Arabidopsis - genetics
Arabidopsis thaliana
Base Sequence
Chromosomes - genetics
Deoxyribonucleic acid
direct DNA uptake
DNA
DNA, Bacterial - genetics
DNA, Plant - genetics
genetic transformation
Genetic Vectors
Infiltration
Molecular Sequence Data
Nicotiana - genetics
Nicotiana plumbaginifolia
Plant Roots - genetics
Plants, Toxic
plasmid vectors
Plasmids - genetics
Protoplasts
Recombination, Genetic
Rhizobium - genetics
roots
Seedlings
Sequence Analysis, DNA
transfer DNA
Transformation, Genetic
vacuum infiltration
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Summary: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 Agiobacterium origin. The high frequency of long transfer after vacuum infiltration of A. thaliana needs to be considered when analyzing T-DNA tagged mutants.
ISSN:0167-4412
1573-5028
DOI:10.1023/a:1005849303333