Level of tissue differentiation influences the activation of a heat-inducible flower-specific system for genetic containment in poplar (Populus tremula L.)

Differentiation level but not transgene copy number influenced activation of a gene containment system in poplar. Heat treatments promoted; CRE; gene body methylation. The flower-specific transgene deletion was confirmed.; Gene flow between genetic modified trees and their wild relatives is still...

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Autores principales: Hoenicka, Hans, Lehnhardt, Denise, Nunna, Suneetha, Reinhardt, Richard, Jeltsch, Albert, Briones, María Valentina, Fladung, Matthias
Formato: Articulo
Lenguaje:Inglés
Publicado: 2015
Materias:
Bioquímica
Ingeniería Agronómica
Biosafety
Populus
Recombination
Gene flow
Heat shock
Acceso en línea:http://sedici.unlp.edu.ar/handle/10915/141484
Aporte de:
SEDICI (UNLP) de Universidad Nacional de La Plata
id I19-R120-10915-141484
record_format dspace
institution Universidad Nacional de La Plata
institution_str I-19
repository_str R-120
collection SEDICI (UNLP)
language Inglés
topic Bioquímica
Ingeniería Agronómica
Biosafety
Populus
Recombination
Gene flow
Heat shock
spellingShingle Bioquímica
Ingeniería Agronómica
Biosafety
Populus
Recombination
Gene flow
Heat shock
Hoenicka, Hans
Lehnhardt, Denise
Nunna, Suneetha
Reinhardt, Richard
Jeltsch, Albert
Briones, María Valentina
Fladung, Matthias
Level of tissue differentiation influences the activation of a heat-inducible flower-specific system for genetic containment in poplar (Populus tremula L.)
topic_facet Bioquímica
Ingeniería Agronómica
Biosafety
Populus
Recombination
Gene flow
Heat shock
description Differentiation level but not transgene copy number influenced activation of a gene containment system in poplar. Heat treatments promoted; CRE; gene body methylation. The flower-specific transgene deletion was confirmed.; 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.
format Articulo
Articulo
author Hoenicka, Hans
Lehnhardt, Denise
Nunna, Suneetha
Reinhardt, Richard
Jeltsch, Albert
Briones, María Valentina
Fladung, Matthias
author_facet Hoenicka, Hans
Lehnhardt, Denise
Nunna, Suneetha
Reinhardt, Richard
Jeltsch, Albert
Briones, María Valentina
Fladung, Matthias
author_sort Hoenicka, Hans
title Level of tissue differentiation influences the activation of a heat-inducible flower-specific system for genetic containment in poplar (Populus tremula L.)
title_short Level of tissue differentiation influences the activation of a heat-inducible flower-specific system for genetic containment in poplar (Populus tremula L.)
title_full Level of tissue differentiation influences the activation of a heat-inducible flower-specific system for genetic containment in poplar (Populus tremula L.)
title_fullStr Level of tissue differentiation influences the activation of a heat-inducible flower-specific system for genetic containment in poplar (Populus tremula L.)
title_full_unstemmed Level of tissue differentiation influences the activation of a heat-inducible flower-specific system for genetic containment in poplar (Populus tremula L.)
title_sort level of tissue differentiation influences the activation of a heat-inducible flower-specific system for genetic containment in poplar (populus tremula l.)
publishDate 2015
url http://sedici.unlp.edu.ar/handle/10915/141484
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