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dc.contributor.author
Hoenicka, Hans
dc.contributor.author
Lehnhardt, Denise
dc.contributor.author
Nunna, Suneetha
dc.contributor.author
Reinhardt, Richard
dc.contributor.author
Jeltsch, Albert
dc.contributor.author
Briones, María Valentina
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dc.contributor.author
Fladung, Matthias
dc.date.available
2019-06-12T21:01:56Z
dc.date.issued
2016-02
dc.identifier.citation
Hoenicka, Hans; Lehnhardt, Denise; Nunna, Suneetha; Reinhardt, Richard; Jeltsch, Albert; et al.; Level of tissue differentiation influences the activation of a heat-inducible flower-specific system for genetic containment in poplar (Populus tremula L.); Springer; Plant Cell Reports; 35; 2; 2-2016; 369-384
dc.identifier.issn
0721-7714
dc.identifier.uri
http://hdl.handle.net/11336/78153
dc.description.abstract
Key message: Differentiation level but not transgene copy number influenced activation of a gene containment system in poplar. Heat treatments promotedCREgene body methylation. The flower-specific transgene deletion was confirmed. Abstract: Gene flow between genetic modified trees and their wild relatives is still motive of concern. Therefore, approaches for gene containment are required. In this study, we designed a novel strategy for achieving an inducible and flower-specific transgene removal from poplar trees but still expressing the transgene in the plant body. Hence, pollen carrying transgenes could be used for breeding purposes under controlled conditions in a first phase, and in the second phase genetic modified poplars developing transgene-free pollen grains could be released. This approach is based on the recombination systems CRE/loxP and FLP/frt. Both gene constructs contained a heat-inducible CRE/loxP-based spacer sequence for in vivo assembling of the flower-specific FLP/frt system. This allowed inducible activation of gene containment. The FLP/frt system was under the regulation of a flower-specific promoter, either CGPDHC or PTD. Our results confirmed complete CRE/loxP-based in vivo assembling of the flower-specific transgene excision system after heat treatment in all cells for up to 30 % of regenerants derived from undifferentiated tissue cultures. Degradation of HSP::CRE/loxP spacer after recombination but also persistence as extrachromosomal DNA circles were detected in sub-lines obtained after heat treatments. Furthermore, heat treatment promoted methylation of the CRE gene body. A lower methylation level was detected at CpG sites in transgenic sub-lines showing complete CRE/loxP recombination and persistence of CRE/loxP spacer, compared to sub-lines with incomplete recombination. However, our results suggest that low methylation might be necessary but not sufficient for recombination. The flower-specific FLP/frt-based transgene deletion was confirmed in 6.3 % of flowers.
dc.format
application/pdf
dc.language.iso
eng
dc.publisher
Springer
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dc.rights
info:eu-repo/semantics/openAccess
dc.rights.uri
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.subject
Biosafety
dc.subject
Gene Flow
dc.subject
Heat Shock
dc.subject
Populus
dc.subject
Recombination
dc.subject.classification
Biología Celular, Microbiología
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dc.subject.classification
Ciencias Biológicas
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dc.subject.classification
CIENCIAS NATURALES Y EXACTAS
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dc.title
Level of tissue differentiation influences the activation of a heat-inducible flower-specific system for genetic containment in poplar (Populus tremula L.)
dc.type
info:eu-repo/semantics/article
dc.type
info:ar-repo/semantics/artículo
dc.type
info:eu-repo/semantics/publishedVersion
dc.date.updated
2019-05-23T19:11:55Z
dc.identifier.eissn
1432-203X
dc.journal.volume
35
dc.journal.number
2
dc.journal.pagination
369-384
dc.journal.pais
Alemania
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dc.journal.ciudad
Berlín
dc.description.fil
Fil: Hoenicka, Hans. Thünen Institut für Forstgenetik; Alemania
dc.description.fil
Fil: Lehnhardt, Denise. Thünen Institut für Forstgenetik; Alemania
dc.description.fil
Fil: Nunna, Suneetha. University of Stuttgart; Alemania
dc.description.fil
Fil: Reinhardt, Richard. Max-Planck-Genome-Centre Cologne; Alemania
dc.description.fil
Fil: Jeltsch, Albert. University of Stuttgart; Alemania
dc.description.fil
Fil: Briones, María Valentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Agrarias y Forestales; Argentina
dc.description.fil
Fil: Fladung, Matthias. Thünen Institut für Forstgenetik; Alemania
dc.journal.title
Plant Cell Reports
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dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/doi/http://dx.doi.org/10.1007/s00299-015-1890-x
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/url/https://link.springer.com/article/10.1007%2Fs00299-015-1890-x
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