A novel soybean hairy root system for gene functional validation.
Autor(a) principal: | |
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Data de Publicação: | 2023 |
Outros Autores: | , , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA - Alice) |
Texto Completo: | http://www.alice.cnptia.embrapa.br/alice/handle/doc/1154066 https://doi. org/10.1371/journal.pone.0285504 |
Resumo: | Agrobacterium rhizogenes-mediated transformation has long been explored as a versatile and reliable method for gene function validation in many plant species, including soybean (Glycine max). Likewise, detached-leaf assays have been widely used for rapid and mass screening of soybean genotypes for disease resistance. The present study combines these two methods to establish an efficient and practical system to generate transgenic soybean hairy roots from detached leaves and their subsequent culture under ex vitro conditions. We demonstrated that hairy roots derived from leaves of two (tropical and temperate) soybean cultivars could be successfully infected by economically important species of root-knot nematodes (Meloidogyne incognita and M. javanica). The established detached-leaf method was further explored for functional validation of two candidate genes encoding for cell wall modifying proteins (CWMPs) to promote resistance against M. incognita through distinct biotechnological strategies: the overexpression of a wild Arachis -expansin transgene (AdEXPA24) and the dsRNA-mediated silencing of an endogenous soybean polygalacturonase gene (GmPG). AdEXPA24 overexpression in hairy roots of RKN-susceptible soybean cultivar significantly reduced nematode infection by approximately 47%, whereas GmPG downregulation caused an average decrease of 37%. This novel system of hairy root induction from detached leaves showed to be an efficient, practical, fast, and low-cost method suitable for high throughput in root analysis of candidate genes in soybean. |
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A novel soybean hairy root system for gene functional validation.Transgenic soybeanAgrobacterium rhizogenesGlycine MaxSojaMelhoramento Genético VegetalMeloidogyne sppSoybeansAgrobacterium rhizogenes-mediated transformation has long been explored as a versatile and reliable method for gene function validation in many plant species, including soybean (Glycine max). Likewise, detached-leaf assays have been widely used for rapid and mass screening of soybean genotypes for disease resistance. The present study combines these two methods to establish an efficient and practical system to generate transgenic soybean hairy roots from detached leaves and their subsequent culture under ex vitro conditions. We demonstrated that hairy roots derived from leaves of two (tropical and temperate) soybean cultivars could be successfully infected by economically important species of root-knot nematodes (Meloidogyne incognita and M. javanica). The established detached-leaf method was further explored for functional validation of two candidate genes encoding for cell wall modifying proteins (CWMPs) to promote resistance against M. incognita through distinct biotechnological strategies: the overexpression of a wild Arachis -expansin transgene (AdEXPA24) and the dsRNA-mediated silencing of an endogenous soybean polygalacturonase gene (GmPG). AdEXPA24 overexpression in hairy roots of RKN-susceptible soybean cultivar significantly reduced nematode infection by approximately 47%, whereas GmPG downregulation caused an average decrease of 37%. This novel system of hairy root induction from detached leaves showed to be an efficient, practical, fast, and low-cost method suitable for high throughput in root analysis of candidate genes in soybean.Na publicação: Maria Fátima Grossi-de-Sa; Patricia Messenberg Guimaraes.BRUNA MEDEIROS PEREIRA; FABRÍCIO ARRAES; ANDRESSA CUNHA QUINTANA MARTINS; NAYARA SABRINA FREITAS ALVES; BRUNO PAES MELO; CAROLINA VIANNA MORGANTE, CPATSA; MARIO ALFREDO PASSOS SARAIVA, Instituto Nacional de Ciência e Tecnologia - INCT; MARIA FATIMA GROSSI DE SA, Cenargen; PATRICIA MESSEMBERG GUIMARAES, Cenargen; ANA CRISTINA MIRANDA BRASILEIRO, Cenargen.PEREIRA, B. M.ARRAES, F.MARTINS, A. C. Q.ALVES, N. S. F.MELO, B. P.MORGANTE, C. V.SARAIVA, M. A. P.SA, M. F. G. deGUIMARAES, P. M.BRASILEIRO, A. C. M.2023-05-29T14:47:59Z2023-05-29T14:47:59Z2023-05-292023info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articlePlos One, v. 18, n. 5, e0285504, 2023.http://www.alice.cnptia.embrapa.br/alice/handle/doc/1154066https://doi. org/10.1371/journal.pone.0285504enginfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA - Alice)instname:Empresa Brasileira de Pesquisa Agropecuária (Embrapa)instacron:EMBRAPA2023-05-29T14:47:59Zoai:www.alice.cnptia.embrapa.br:doc/1154066Repositório InstitucionalPUBhttps://www.alice.cnptia.embrapa.br/oai/requestopendoar:21542023-05-29T14:47:59falseRepositório InstitucionalPUBhttps://www.alice.cnptia.embrapa.br/oai/requestcg-riaa@embrapa.bropendoar:21542023-05-29T14:47:59Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA - Alice) - Empresa Brasileira de Pesquisa Agropecuária (Embrapa)false |
dc.title.none.fl_str_mv |
A novel soybean hairy root system for gene functional validation. |
title |
A novel soybean hairy root system for gene functional validation. |
spellingShingle |
A novel soybean hairy root system for gene functional validation. PEREIRA, B. M. Transgenic soybean Agrobacterium rhizogenes Glycine Max Soja Melhoramento Genético Vegetal Meloidogyne spp Soybeans |
title_short |
A novel soybean hairy root system for gene functional validation. |
title_full |
A novel soybean hairy root system for gene functional validation. |
title_fullStr |
A novel soybean hairy root system for gene functional validation. |
title_full_unstemmed |
A novel soybean hairy root system for gene functional validation. |
title_sort |
A novel soybean hairy root system for gene functional validation. |
author |
PEREIRA, B. M. |
author_facet |
PEREIRA, B. M. ARRAES, F. MARTINS, A. C. Q. ALVES, N. S. F. MELO, B. P. MORGANTE, C. V. SARAIVA, M. A. P. SA, M. F. G. de GUIMARAES, P. M. BRASILEIRO, A. C. M. |
author_role |
author |
author2 |
ARRAES, F. MARTINS, A. C. Q. ALVES, N. S. F. MELO, B. P. MORGANTE, C. V. SARAIVA, M. A. P. SA, M. F. G. de GUIMARAES, P. M. BRASILEIRO, A. C. M. |
author2_role |
author author author author author author author author author |
dc.contributor.none.fl_str_mv |
BRUNA MEDEIROS PEREIRA; FABRÍCIO ARRAES; ANDRESSA CUNHA QUINTANA MARTINS; NAYARA SABRINA FREITAS ALVES; BRUNO PAES MELO; CAROLINA VIANNA MORGANTE, CPATSA; MARIO ALFREDO PASSOS SARAIVA, Instituto Nacional de Ciência e Tecnologia - INCT; MARIA FATIMA GROSSI DE SA, Cenargen; PATRICIA MESSEMBERG GUIMARAES, Cenargen; ANA CRISTINA MIRANDA BRASILEIRO, Cenargen. |
dc.contributor.author.fl_str_mv |
PEREIRA, B. M. ARRAES, F. MARTINS, A. C. Q. ALVES, N. S. F. MELO, B. P. MORGANTE, C. V. SARAIVA, M. A. P. SA, M. F. G. de GUIMARAES, P. M. BRASILEIRO, A. C. M. |
dc.subject.por.fl_str_mv |
Transgenic soybean Agrobacterium rhizogenes Glycine Max Soja Melhoramento Genético Vegetal Meloidogyne spp Soybeans |
topic |
Transgenic soybean Agrobacterium rhizogenes Glycine Max Soja Melhoramento Genético Vegetal Meloidogyne spp Soybeans |
description |
Agrobacterium rhizogenes-mediated transformation has long been explored as a versatile and reliable method for gene function validation in many plant species, including soybean (Glycine max). Likewise, detached-leaf assays have been widely used for rapid and mass screening of soybean genotypes for disease resistance. The present study combines these two methods to establish an efficient and practical system to generate transgenic soybean hairy roots from detached leaves and their subsequent culture under ex vitro conditions. We demonstrated that hairy roots derived from leaves of two (tropical and temperate) soybean cultivars could be successfully infected by economically important species of root-knot nematodes (Meloidogyne incognita and M. javanica). The established detached-leaf method was further explored for functional validation of two candidate genes encoding for cell wall modifying proteins (CWMPs) to promote resistance against M. incognita through distinct biotechnological strategies: the overexpression of a wild Arachis -expansin transgene (AdEXPA24) and the dsRNA-mediated silencing of an endogenous soybean polygalacturonase gene (GmPG). AdEXPA24 overexpression in hairy roots of RKN-susceptible soybean cultivar significantly reduced nematode infection by approximately 47%, whereas GmPG downregulation caused an average decrease of 37%. This novel system of hairy root induction from detached leaves showed to be an efficient, practical, fast, and low-cost method suitable for high throughput in root analysis of candidate genes in soybean. |
publishDate |
2023 |
dc.date.none.fl_str_mv |
2023-05-29T14:47:59Z 2023-05-29T14:47:59Z 2023-05-29 2023 |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/publishedVersion info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
Plos One, v. 18, n. 5, e0285504, 2023. http://www.alice.cnptia.embrapa.br/alice/handle/doc/1154066 https://doi. org/10.1371/journal.pone.0285504 |
identifier_str_mv |
Plos One, v. 18, n. 5, e0285504, 2023. |
url |
http://www.alice.cnptia.embrapa.br/alice/handle/doc/1154066 https://doi. org/10.1371/journal.pone.0285504 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.source.none.fl_str_mv |
reponame:Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA - Alice) instname:Empresa Brasileira de Pesquisa Agropecuária (Embrapa) instacron:EMBRAPA |
instname_str |
Empresa Brasileira de Pesquisa Agropecuária (Embrapa) |
instacron_str |
EMBRAPA |
institution |
EMBRAPA |
reponame_str |
Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA - Alice) |
collection |
Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA - Alice) |
repository.name.fl_str_mv |
Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA - Alice) - Empresa Brasileira de Pesquisa Agropecuária (Embrapa) |
repository.mail.fl_str_mv |
cg-riaa@embrapa.br |
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1794503545213419520 |