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Transgene autoexcision in switchgrass pollen mediated by the Bxb1 recombinase

BACKGROUND: Switchgrass (Panicum virgatum L.) has a great potential as a platform for the production of biobased plastics, chemicals and energy mainly because of its high biomass yield on marginal land and low agricultural inputs. During the last decade, there has been increased interest in the gene...

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Autores principales: Somleva, Maria N, Xu, Chang Ai, Ryan, Kieran P, Thilmony, Roger, Peoples, Oliver, Snell, Kristi D, Thomson, James
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4148497/
https://www.ncbi.nlm.nih.gov/pubmed/25148894
http://dx.doi.org/10.1186/1472-6750-14-79
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author Somleva, Maria N
Xu, Chang Ai
Ryan, Kieran P
Thilmony, Roger
Peoples, Oliver
Snell, Kristi D
Thomson, James
author_facet Somleva, Maria N
Xu, Chang Ai
Ryan, Kieran P
Thilmony, Roger
Peoples, Oliver
Snell, Kristi D
Thomson, James
author_sort Somleva, Maria N
collection PubMed
description BACKGROUND: Switchgrass (Panicum virgatum L.) has a great potential as a platform for the production of biobased plastics, chemicals and energy mainly because of its high biomass yield on marginal land and low agricultural inputs. During the last decade, there has been increased interest in the genetic improvement of this crop through transgenic approaches. Since switchgrass, like most perennial grasses, is exclusively cross pollinating and poorly domesticated, preventing the dispersal of transgenic pollen into the environment is a critical requisite for the commercial deployment of this important biomass crop. In this study, the feasibility of controlling pollen-mediated gene flow in transgenic switchgrass using the large serine site-specific recombinase Bxb1 has been investigated. RESULTS: A novel approach utilizing co-transformation of two separate vectors was used to test the functionality of the Bxb1/att recombination system in switchgrass. In addition, two promoters with high pollen-specific activity were identified and thoroughly characterized prior to their introduction into a test vector explicitly designed for both autoexcision and quantitative analyses of recombination events. Our strategy for developmentally programmed precise excision of the recombinase and marker genes in switchgrass pollen resulted in the generation of transgene-excised progeny. The autoexcision efficiencies were in the range of 22-42% depending on the transformation event and assay used. CONCLUSION: The results presented here mark an important milestone towards the establishment of a reliable biocontainment system for switchgrass which will facilitate the development of this crop as a biorefinery feedstock through advanced biotechnological approaches.
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spelling pubmed-41484972014-08-30 Transgene autoexcision in switchgrass pollen mediated by the Bxb1 recombinase Somleva, Maria N Xu, Chang Ai Ryan, Kieran P Thilmony, Roger Peoples, Oliver Snell, Kristi D Thomson, James BMC Biotechnol Research Article BACKGROUND: Switchgrass (Panicum virgatum L.) has a great potential as a platform for the production of biobased plastics, chemicals and energy mainly because of its high biomass yield on marginal land and low agricultural inputs. During the last decade, there has been increased interest in the genetic improvement of this crop through transgenic approaches. Since switchgrass, like most perennial grasses, is exclusively cross pollinating and poorly domesticated, preventing the dispersal of transgenic pollen into the environment is a critical requisite for the commercial deployment of this important biomass crop. In this study, the feasibility of controlling pollen-mediated gene flow in transgenic switchgrass using the large serine site-specific recombinase Bxb1 has been investigated. RESULTS: A novel approach utilizing co-transformation of two separate vectors was used to test the functionality of the Bxb1/att recombination system in switchgrass. In addition, two promoters with high pollen-specific activity were identified and thoroughly characterized prior to their introduction into a test vector explicitly designed for both autoexcision and quantitative analyses of recombination events. Our strategy for developmentally programmed precise excision of the recombinase and marker genes in switchgrass pollen resulted in the generation of transgene-excised progeny. The autoexcision efficiencies were in the range of 22-42% depending on the transformation event and assay used. CONCLUSION: The results presented here mark an important milestone towards the establishment of a reliable biocontainment system for switchgrass which will facilitate the development of this crop as a biorefinery feedstock through advanced biotechnological approaches. BioMed Central 2014-08-22 /pmc/articles/PMC4148497/ /pubmed/25148894 http://dx.doi.org/10.1186/1472-6750-14-79 Text en Copyright © 2014 Somleva et al.; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/4.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research Article
Somleva, Maria N
Xu, Chang Ai
Ryan, Kieran P
Thilmony, Roger
Peoples, Oliver
Snell, Kristi D
Thomson, James
Transgene autoexcision in switchgrass pollen mediated by the Bxb1 recombinase
title Transgene autoexcision in switchgrass pollen mediated by the Bxb1 recombinase
title_full Transgene autoexcision in switchgrass pollen mediated by the Bxb1 recombinase
title_fullStr Transgene autoexcision in switchgrass pollen mediated by the Bxb1 recombinase
title_full_unstemmed Transgene autoexcision in switchgrass pollen mediated by the Bxb1 recombinase
title_short Transgene autoexcision in switchgrass pollen mediated by the Bxb1 recombinase
title_sort transgene autoexcision in switchgrass pollen mediated by the bxb1 recombinase
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4148497/
https://www.ncbi.nlm.nih.gov/pubmed/25148894
http://dx.doi.org/10.1186/1472-6750-14-79
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