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Capture of genomic and T‐DNA sequences during double‐strand break repair in somatic plant cells
Author(s) -
Salomon Siegfried,
Puchta Holger
Publication year - 1998
Publication title -
the embo journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.484
H-Index - 392
eISSN - 1460-2075
pISSN - 0261-4189
DOI - 10.1093/emboj/17.20.6086
Subject(s) - biology , somatic cell , genetics , dna , dna repair , microbiology and biotechnology , computational biology , gene
To analyze genomic changes resulting from double‐strand break (DSB) repair, transgenic tobacco plants were obtained that carried in their genome a restriction site of the rare cutting endonuclease I‐ Sce I within a negative selectable marker gene. After induction of DSB repair via Agrobacterium ‐mediated transient expression of I‐ Sce I, plant cells were selected that carried a loss‐of‐function phenotype of the marker. Surprisingly, in addition to deletions, in a number of cases repair was associated with the insertion of unique and repetitive genomic sequences into the break. Thus, DSB repair offers a mechanism for spreading different kinds of sequences into new chromosomal positions. This may have evolutionary consequences particularly for plants, as genomic alterations occurring in meristem cells can be transferred to the next generation. Moreover, transfer DNA (T‐DNA), carrying the open reading frame of I‐ Sce I, was found in several cases to be integrated into the transgenic I‐ Sce I site. This indicates that DSB repair also represents a pathway for the integration of T‐DNA into the plant genome.