pGVG: a new Gateway-compatible vector for transformation of sugarcane and other monocot crops
Autor(a) principal: | |
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Data de Publicação: | 2018 |
Outros Autores: | , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Genetics and Molecular Biology |
Texto Completo: | http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572018000300450 |
Resumo: | Abstract The successful development of genetically engineered monocots using Agrobacterium-mediated transformation has created an increasing demand for compatible vectors. We have developed a new expression vector, pGVG, for efficient transformation and expression of different constructs for gene overexpression and silencing in sugarcane. The pCAMBIA2300 binary vector was modified by adding Gateway recombination sites for fast gene transfer between vectors and the maize polyubiquitin promoter Ubi-1 (ZmUbi1), which is known to drive high gene expression levels in monocots. Transformation efficiency using the pGVG vector reached up to 14 transgenic events per gram of transformed callus. Transgenic plants expressing the β-glucuronidase (GUS) reporter gene from pGVG showed high levels of GUS activity. qRT-PCR evaluations demonstrated success for both overexpression and hairpin-based silencing cassettes. Therefore, pGVG is suitable for plant transformation and subsequent applications for high-throughput production of stable transgenic sugarcane. The use of an expression cassette based on the ZmUbi1 promoter opens the possibility of using pGVG in other monocot species. |
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Genetics and Molecular Biology |
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pGVG: a new Gateway-compatible vector for transformation of sugarcane and other monocot cropsMonocotssugarcanevectorGateway technologygenetic transformationAbstract The successful development of genetically engineered monocots using Agrobacterium-mediated transformation has created an increasing demand for compatible vectors. We have developed a new expression vector, pGVG, for efficient transformation and expression of different constructs for gene overexpression and silencing in sugarcane. The pCAMBIA2300 binary vector was modified by adding Gateway recombination sites for fast gene transfer between vectors and the maize polyubiquitin promoter Ubi-1 (ZmUbi1), which is known to drive high gene expression levels in monocots. Transformation efficiency using the pGVG vector reached up to 14 transgenic events per gram of transformed callus. Transgenic plants expressing the β-glucuronidase (GUS) reporter gene from pGVG showed high levels of GUS activity. qRT-PCR evaluations demonstrated success for both overexpression and hairpin-based silencing cassettes. Therefore, pGVG is suitable for plant transformation and subsequent applications for high-throughput production of stable transgenic sugarcane. The use of an expression cassette based on the ZmUbi1 promoter opens the possibility of using pGVG in other monocot species.Sociedade Brasileira de Genética2018-06-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572018000300450Genetics and Molecular Biology v.41 n.2 2018reponame:Genetics and Molecular Biologyinstname:Sociedade Brasileira de Genética (SBG)instacron:SBG10.1590/1678-4685-gmb-2017-0262info:eu-repo/semantics/openAccessGuidelli,Giovanna V.Mattiello,LuciaGallinari,Rafael H.Lucca,Paulo Cezar deMenossi,Marceloeng2018-06-21T00:00:00Zoai:scielo:S1415-47572018000300450Revistahttp://www.gmb.org.br/ONGhttps://old.scielo.br/oai/scielo-oai.php||editor@gmb.org.br1678-46851415-4757opendoar:2018-06-21T00:00Genetics and Molecular Biology - Sociedade Brasileira de Genética (SBG)false |
dc.title.none.fl_str_mv |
pGVG: a new Gateway-compatible vector for transformation of sugarcane and other monocot crops |
title |
pGVG: a new Gateway-compatible vector for transformation of sugarcane and other monocot crops |
spellingShingle |
pGVG: a new Gateway-compatible vector for transformation of sugarcane and other monocot crops Guidelli,Giovanna V. Monocots sugarcane vector Gateway technology genetic transformation |
title_short |
pGVG: a new Gateway-compatible vector for transformation of sugarcane and other monocot crops |
title_full |
pGVG: a new Gateway-compatible vector for transformation of sugarcane and other monocot crops |
title_fullStr |
pGVG: a new Gateway-compatible vector for transformation of sugarcane and other monocot crops |
title_full_unstemmed |
pGVG: a new Gateway-compatible vector for transformation of sugarcane and other monocot crops |
title_sort |
pGVG: a new Gateway-compatible vector for transformation of sugarcane and other monocot crops |
author |
Guidelli,Giovanna V. |
author_facet |
Guidelli,Giovanna V. Mattiello,Lucia Gallinari,Rafael H. Lucca,Paulo Cezar de Menossi,Marcelo |
author_role |
author |
author2 |
Mattiello,Lucia Gallinari,Rafael H. Lucca,Paulo Cezar de Menossi,Marcelo |
author2_role |
author author author author |
dc.contributor.author.fl_str_mv |
Guidelli,Giovanna V. Mattiello,Lucia Gallinari,Rafael H. Lucca,Paulo Cezar de Menossi,Marcelo |
dc.subject.por.fl_str_mv |
Monocots sugarcane vector Gateway technology genetic transformation |
topic |
Monocots sugarcane vector Gateway technology genetic transformation |
description |
Abstract The successful development of genetically engineered monocots using Agrobacterium-mediated transformation has created an increasing demand for compatible vectors. We have developed a new expression vector, pGVG, for efficient transformation and expression of different constructs for gene overexpression and silencing in sugarcane. The pCAMBIA2300 binary vector was modified by adding Gateway recombination sites for fast gene transfer between vectors and the maize polyubiquitin promoter Ubi-1 (ZmUbi1), which is known to drive high gene expression levels in monocots. Transformation efficiency using the pGVG vector reached up to 14 transgenic events per gram of transformed callus. Transgenic plants expressing the β-glucuronidase (GUS) reporter gene from pGVG showed high levels of GUS activity. qRT-PCR evaluations demonstrated success for both overexpression and hairpin-based silencing cassettes. Therefore, pGVG is suitable for plant transformation and subsequent applications for high-throughput production of stable transgenic sugarcane. The use of an expression cassette based on the ZmUbi1 promoter opens the possibility of using pGVG in other monocot species. |
publishDate |
2018 |
dc.date.none.fl_str_mv |
2018-06-01 |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572018000300450 |
url |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572018000300450 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
10.1590/1678-4685-gmb-2017-0262 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
text/html |
dc.publisher.none.fl_str_mv |
Sociedade Brasileira de Genética |
publisher.none.fl_str_mv |
Sociedade Brasileira de Genética |
dc.source.none.fl_str_mv |
Genetics and Molecular Biology v.41 n.2 2018 reponame:Genetics and Molecular Biology instname:Sociedade Brasileira de Genética (SBG) instacron:SBG |
instname_str |
Sociedade Brasileira de Genética (SBG) |
instacron_str |
SBG |
institution |
SBG |
reponame_str |
Genetics and Molecular Biology |
collection |
Genetics and Molecular Biology |
repository.name.fl_str_mv |
Genetics and Molecular Biology - Sociedade Brasileira de Genética (SBG) |
repository.mail.fl_str_mv |
||editor@gmb.org.br |
_version_ |
1752122388822097920 |