PoplarV1, Main, Exploration, bibRecord, 002978

One-step Agrobacterium mediated transformation of eight genes essential for rhizobium symbiotic signaling using the novel binary vector system pHUGE.

Identifieur interne : 002978 ( Main/Exploration ); précédent : 002977; suivant : 002979

One-step Agrobacterium mediated transformation of eight genes essential for rhizobium symbiotic signaling using the novel binary vector system pHUGE.

Auteurs : Andreas Untergasser [Pays-Bas] ; Gerben J M. Bijl ; Wei Liu ; Ton Bisseling ; Jan G. Schaart ; René Geurts

Source :

PloS one [ 1932-6203 ] ; 2012.

RBID : pubmed:23112864

Descripteurs français

KwdFr :
- ADN bactérien (génétique), Agrobacterium tumefaciens (génétique), Clonage moléculaire (méthodes), Medicago truncatula (génétique), Medicago truncatula (microbiologie), Plantes (génétique), Plantes (microbiologie), Rhizobium (physiologie), Symbiose (MeSH), Transformation génétique (MeSH), Transgènes (MeSH), Vecteurs génétiques (génétique), Végétaux génétiquement modifiés (génétique), Végétaux génétiquement modifiés (microbiologie).
MESH :
- génétique : ADN bactérien, Agrobacterium tumefaciens, Medicago truncatula, Plantes, Vecteurs génétiques, Végétaux génétiquement modifiés.
- microbiologie : Medicago truncatula, Plantes, Végétaux génétiquement modifiés.
- méthodes : Clonage moléculaire.
- physiologie : Rhizobium.
- Symbiose, Transformation génétique, Transgènes.

English descriptors

KwdEn :
- Agrobacterium tumefaciens (genetics), Cloning, Molecular (methods), DNA, Bacterial (genetics), Genetic Vectors (genetics), Medicago truncatula (genetics), Medicago truncatula (microbiology), Plants (genetics), Plants (microbiology), Plants, Genetically Modified (genetics), Plants, Genetically Modified (microbiology), Rhizobium (physiology), Symbiosis (MeSH), Transformation, Genetic (MeSH), Transgenes (MeSH).
MESH :
- chemical , genetics : DNA, Bacterial.
- genetics : Agrobacterium tumefaciens, Genetic Vectors, Medicago truncatula, Plants, Plants, Genetically Modified.
- methods : Cloning, Molecular.
- microbiology : Medicago truncatula, Plants, Plants, Genetically Modified.
- physiology : Rhizobium.
- Symbiosis, Transformation, Genetic, Transgenes.

Abstract

Advancement in plant research is becoming impaired by the fact that the transfer of multiple genes is difficult to achieve. Here we present a new binary vector for Agrobacterium tumefaciens mediated transformation, pHUGE-Red, in concert with a cloning strategy suited for the transfer of up to nine genes at once. This vector enables modular cloning of large DNA fragments by employing Gateway technology and contains DsRED1 as visual selection marker. Furthermore, an R/Rs inducible recombination system was included allowing subsequent removal of the selection markers in the newly generated transgenic plants. We show the successful use of pHUGE-Red by transferring eight genes essential for Medicago truncatula to establish a symbiosis with rhizobia bacteria as one 74 kb T-DNA into four non-leguminous species; strawberry, poplar, tomato and tobacco. We provide evidence that all transgenes are expressed in the root tissue of the non-legumes. Visual control during the transformation process and subsequent marker gene removal makes the pHUGE-Red vector an excellent tool for the efficient transfer of multiple genes.

DOI: 10.1371/journal.pone.0047885
PubMed: 23112864
PubMed Central: PMC3480454

Affiliations:

Links toward previous steps (curation, corpus...)

to stream Main, to step Corpus: 002828
to stream Main, to step Curation: 002828

Le document en format XML

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<term>Genetic Vectors (genetics)</term>
<term>Medicago truncatula (genetics)</term>
<term>Medicago truncatula (microbiology)</term>
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<term>Plants, Genetically Modified (microbiology)</term>
<term>Rhizobium (physiology)</term>
<term>Symbiosis (MeSH)</term>
<term>Transformation, Genetic (MeSH)</term>
<term>Transgenes (MeSH)</term>
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<term>Agrobacterium tumefaciens (génétique)</term>
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<term>Medicago truncatula (génétique)</term>
<term>Medicago truncatula (microbiologie)</term>
<term>Plantes (génétique)</term>
<term>Plantes (microbiologie)</term>
<term>Rhizobium (physiologie)</term>
<term>Symbiose (MeSH)</term>
<term>Transformation génétique (MeSH)</term>
<term>Transgènes (MeSH)</term>
<term>Vecteurs génétiques (génétique)</term>
<term>Végétaux génétiquement modifiés (génétique)</term>
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<term>Vecteurs génétiques</term>
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<term>Transgenes</term>
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<front><div type="abstract" xml:lang="en">Advancement in plant research is becoming impaired by the fact that the transfer of multiple genes is difficult to achieve. Here we present a new binary vector for Agrobacterium tumefaciens mediated transformation, pHUGE-Red, in concert with a cloning strategy suited for the transfer of up to nine genes at once. This vector enables modular cloning of large DNA fragments by employing Gateway technology and contains DsRED1 as visual selection marker. Furthermore, an R/Rs inducible recombination system was included allowing subsequent removal of the selection markers in the newly generated transgenic plants. We show the successful use of pHUGE-Red by transferring eight genes essential for Medicago truncatula to establish a symbiosis with rhizobia bacteria as one 74 kb T-DNA into four non-leguminous species; strawberry, poplar, tomato and tobacco. We provide evidence that all transgenes are expressed in the root tissue of the non-legumes. Visual control during the transformation process and subsequent marker gene removal makes the pHUGE-Red vector an excellent tool for the efficient transfer of multiple genes.</div>
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One-step Agrobacterium mediated transformation of eight genes essential for rhizobium symbiotic signaling using the novel binary vector system pHUGE.

One-step Agrobacterium mediated transformation of eight genes essential for rhizobium symbiotic signaling using the novel binary vector system pHUGE.

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