Methods for Cercospora coffeicola protoplast isolation and genetic transformation with the green fluorescent protein

Detalhes bibliográficos
Autor(a) principal: Souza, André Gomes Coelho
Data de Publicação: 2014
Outros Autores: Herrero, Sonia, Maffia, Luiz Antonio, Daub, Margaret Elizabeth
Tipo de documento: Artigo
Idioma: eng
Título da fonte: LOCUS Repositório Institucional da UFV
Texto Completo: https://doi.org/10.1007/s10658-013-0301-9
http://www.locus.ufv.br/handle/123456789/24076
Resumo: Cercospora coffeicola is the causal agent of brown eye spot on coffee leaves. Although the disease has significant importance, few molecular studies have been done with C. coffeicola. Here we report a protocol for isolating protoplasts as well as development of a genetic transformation system using Green Fluorescent Protein. High yields of protoplasts (≈108/ml) were obtained from mycelial cultures from five isolates of C. coffeicola. One isolate was transformed with a vector encoding hygromycin resistance and Green Fluorescent Protein. Out of 43 hygromycin-resistant transformants obtained, Green Fluorescent Protein was highly expressed in one (2.3 %).
id UFV_a36ae5c0f3fe421b2473b18876afed04
oai_identifier_str oai:locus.ufv.br:123456789/24076
network_acronym_str UFV
network_name_str LOCUS Repositório Institucional da UFV
repository_id_str 2145
spelling Souza, André Gomes CoelhoHerrero, SoniaMaffia, Luiz AntonioDaub, Margaret Elizabeth2019-03-21T18:10:13Z2019-03-21T18:10:13Z2014-061573-8469https://doi.org/10.1007/s10658-013-0301-9http://www.locus.ufv.br/handle/123456789/24076Cercospora coffeicola is the causal agent of brown eye spot on coffee leaves. Although the disease has significant importance, few molecular studies have been done with C. coffeicola. Here we report a protocol for isolating protoplasts as well as development of a genetic transformation system using Green Fluorescent Protein. High yields of protoplasts (≈108/ml) were obtained from mycelial cultures from five isolates of C. coffeicola. One isolate was transformed with a vector encoding hygromycin resistance and Green Fluorescent Protein. Out of 43 hygromycin-resistant transformants obtained, Green Fluorescent Protein was highly expressed in one (2.3 %).engEuropean Journal of Plant PathologyVolume 139, Issue 2, Pages 241–244, June 2014KNPVinfo:eu-repo/semantics/openAccessBrown eye spotCoffea arabicaL. Leaf spotMethods for Cercospora coffeicola protoplast isolation and genetic transformation with the green fluorescent proteininfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfreponame:LOCUS Repositório Institucional da UFVinstname:Universidade Federal de Viçosa (UFV)instacron:UFVORIGINALartigo.pdfartigo.pdfTexto completoapplication/pdf285549https://locus.ufv.br//bitstream/123456789/24076/1/artigo.pdfb1ef21bcbbe7803c3576524d85dd6913MD51LICENSElicense.txtlicense.txttext/plain; charset=utf-81748https://locus.ufv.br//bitstream/123456789/24076/2/license.txt8a4605be74aa9ea9d79846c1fba20a33MD52123456789/240762019-03-21 15:13:35.332oai:locus.ufv.br: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Repositório InstitucionalPUBhttps://www.locus.ufv.br/oai/requestfabiojreis@ufv.bropendoar:21452019-03-21T18:13:35LOCUS Repositório Institucional da UFV - Universidade Federal de Viçosa (UFV)false
dc.title.en.fl_str_mv Methods for Cercospora coffeicola protoplast isolation and genetic transformation with the green fluorescent protein
title Methods for Cercospora coffeicola protoplast isolation and genetic transformation with the green fluorescent protein
spellingShingle Methods for Cercospora coffeicola protoplast isolation and genetic transformation with the green fluorescent protein
Souza, André Gomes Coelho
Brown eye spot
Coffea arabica
L. Leaf spot
title_short Methods for Cercospora coffeicola protoplast isolation and genetic transformation with the green fluorescent protein
title_full Methods for Cercospora coffeicola protoplast isolation and genetic transformation with the green fluorescent protein
title_fullStr Methods for Cercospora coffeicola protoplast isolation and genetic transformation with the green fluorescent protein
title_full_unstemmed Methods for Cercospora coffeicola protoplast isolation and genetic transformation with the green fluorescent protein
title_sort Methods for Cercospora coffeicola protoplast isolation and genetic transformation with the green fluorescent protein
author Souza, André Gomes Coelho
author_facet Souza, André Gomes Coelho
Herrero, Sonia
Maffia, Luiz Antonio
Daub, Margaret Elizabeth
author_role author
author2 Herrero, Sonia
Maffia, Luiz Antonio
Daub, Margaret Elizabeth
author2_role author
author
author
dc.contributor.author.fl_str_mv Souza, André Gomes Coelho
Herrero, Sonia
Maffia, Luiz Antonio
Daub, Margaret Elizabeth
dc.subject.pt-BR.fl_str_mv Brown eye spot
Coffea arabica
L. Leaf spot
topic Brown eye spot
Coffea arabica
L. Leaf spot
description Cercospora coffeicola is the causal agent of brown eye spot on coffee leaves. Although the disease has significant importance, few molecular studies have been done with C. coffeicola. Here we report a protocol for isolating protoplasts as well as development of a genetic transformation system using Green Fluorescent Protein. High yields of protoplasts (≈108/ml) were obtained from mycelial cultures from five isolates of C. coffeicola. One isolate was transformed with a vector encoding hygromycin resistance and Green Fluorescent Protein. Out of 43 hygromycin-resistant transformants obtained, Green Fluorescent Protein was highly expressed in one (2.3 %).
publishDate 2014
dc.date.issued.fl_str_mv 2014-06
dc.date.accessioned.fl_str_mv 2019-03-21T18:10:13Z
dc.date.available.fl_str_mv 2019-03-21T18:10:13Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv https://doi.org/10.1007/s10658-013-0301-9
http://www.locus.ufv.br/handle/123456789/24076
dc.identifier.issn.none.fl_str_mv 1573-8469
identifier_str_mv 1573-8469
url https://doi.org/10.1007/s10658-013-0301-9
http://www.locus.ufv.br/handle/123456789/24076
dc.language.iso.fl_str_mv eng
language eng
dc.relation.ispartofseries.pt-BR.fl_str_mv Volume 139, Issue 2, Pages 241–244, June 2014
dc.rights.driver.fl_str_mv KNPV
info:eu-repo/semantics/openAccess
rights_invalid_str_mv KNPV
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv European Journal of Plant Pathology
publisher.none.fl_str_mv European Journal of Plant Pathology
dc.source.none.fl_str_mv reponame:LOCUS Repositório Institucional da UFV
instname:Universidade Federal de Viçosa (UFV)
instacron:UFV
instname_str Universidade Federal de Viçosa (UFV)
instacron_str UFV
institution UFV
reponame_str LOCUS Repositório Institucional da UFV
collection LOCUS Repositório Institucional da UFV
bitstream.url.fl_str_mv https://locus.ufv.br//bitstream/123456789/24076/1/artigo.pdf
https://locus.ufv.br//bitstream/123456789/24076/2/license.txt
bitstream.checksum.fl_str_mv b1ef21bcbbe7803c3576524d85dd6913
8a4605be74aa9ea9d79846c1fba20a33
bitstream.checksumAlgorithm.fl_str_mv MD5
MD5
repository.name.fl_str_mv LOCUS Repositório Institucional da UFV - Universidade Federal de Viçosa (UFV)
repository.mail.fl_str_mv fabiojreis@ufv.br
_version_ 1801212989854973952

Registros relacionados