An Insight into T-DNA Integration Events in Medicago sativa

The molecular mechanisms of transferred DNA (T-DNA) integration into the plant genome are still not completely understood. A large number of integration events have been analyzed in different species, shedding light on the molecular mechanisms involved, and on the frequent transfer of vector sequenc...

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Bibliographic Details
Published in:International journal of molecular sciences 2017-09, Vol.18 (9), p.1951
Main Authors: Nicolia, Alessandro, Ferradini, Nicoletta, Veronesi, Fabio, Rosellini, Daniele
Format: Article
Language:English
Subjects:
Agrobacterium tumefaciens
Agrobacterium tumefaciens - genetics
Alfalfa
Base Sequence
Deoxyribonucleic acid
DNA
DNA, Bacterial - genetics
DNA, Plant - genetics
Genetic Engineering
Genetic Vectors
Genome, Plant
Genomes
Integration
Medicago sativa
Medicago sativa - genetics
Molecular modelling
Nucleotide sequence
Plants, Genetically Modified - genetics
Sequence Analysis, DNA
T-DNA
T-DNA integration
Transformation, Genetic - genetics
Transgenic plants
vector backbone
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Summary:The molecular mechanisms of transferred DNA (T-DNA) integration into the plant genome are still not completely understood. A large number of integration events have been analyzed in different species, shedding light on the molecular mechanisms involved, and on the frequent transfer of vector sequences outside the T-DNA borders, the so-called vector backbone (VB) sequences. In this work, we characterized 46 transgenic alfalfa ( L.) plants (events), generated in previous works, for the presence of VB tracts, and sequenced several T-DNA/genomic DNA (gDNA) junctions. We observed that about 29% of the transgenic events contained VB sequences, within the range reported in other species. Sequence analysis of the T-DNA/gDNA junctions evidenced larger deletions at LBs compared to RBs and insertions probably originated by different integration mechanisms. Overall, our findings in alfalfa are consistent with those in other plant species. This work extends the knowledge on the molecular events of T-DNA integration and can help to design better transformation protocols for alfalfa.
ISSN:1422-0067
1661-6596
1422-0067
DOI:10.3390/ijms18091951