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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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Autores principales: Nicolia, Alessandro, Ferradini, Nicoletta, Veronesi, Fabio, Rosellini, Daniele
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5618600/
https://www.ncbi.nlm.nih.gov/pubmed/28895894
http://dx.doi.org/10.3390/ijms18091951
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author Nicolia, Alessandro
Ferradini, Nicoletta
Veronesi, Fabio
Rosellini, Daniele
author_facet Nicolia, Alessandro
Ferradini, Nicoletta
Veronesi, Fabio
Rosellini, Daniele
author_sort Nicolia, Alessandro
collection PubMed
description 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 (Medicago sativa 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.
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spelling pubmed-56186002017-09-30 An Insight into T-DNA Integration Events in Medicago sativa Nicolia, Alessandro Ferradini, Nicoletta Veronesi, Fabio Rosellini, Daniele Int J Mol Sci Article 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 (Medicago sativa 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. MDPI 2017-09-12 /pmc/articles/PMC5618600/ /pubmed/28895894 http://dx.doi.org/10.3390/ijms18091951 Text en © 2017 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Nicolia, Alessandro
Ferradini, Nicoletta
Veronesi, Fabio
Rosellini, Daniele
An Insight into T-DNA Integration Events in Medicago sativa
title An Insight into T-DNA Integration Events in Medicago sativa
title_full An Insight into T-DNA Integration Events in Medicago sativa
title_fullStr An Insight into T-DNA Integration Events in Medicago sativa
title_full_unstemmed An Insight into T-DNA Integration Events in Medicago sativa
title_short An Insight into T-DNA Integration Events in Medicago sativa
title_sort insight into t-dna integration events in medicago sativa
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5618600/
https://www.ncbi.nlm.nih.gov/pubmed/28895894
http://dx.doi.org/10.3390/ijms18091951
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