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Registro Completo |
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Biblioteca(s): |
Embrapa Recursos Genéticos e Biotecnologia; Embrapa Semiárido. |
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Data corrente: |
13/09/2021 |
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Data da última atualização: |
15/09/2021 |
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Tipo da produção científica: |
Artigo em Periódico Indexado |
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Autoria: |
RIBEIRO, T. P.; LOURENCO, I. T.; MELO, B. P. de; MORGANTE, C. V.; SALLES FILHO, A.; LINS, C. B. J.; FERREIRA, G. F.; MELLO, G. N.; MACEDO, L. L. P. de; LUCENA, W. A.; SILVA, M. C. M. da; OLIVEIRA‑NETO, O. B.; SA, M. F. G. de. |
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Afiliação: |
THUANNE PIRES RIBEIRO, UNB; ISABELA TRISTAN LOURENCO TESSUTTI, Cenargen; BRUNO PAES DE MELO, UFV; CAROLINA VIANNA MORGANTE, CPATSA; ALVARO SALLES FILHO, UCB; CAMILA BARROZO JESUS LINS; GILANNA FALCÃO FERREIRA; GLÊNIA NUNES MELLO; LEONARDO LIMA PEPINO DE MACEDO, Cenargen; WAGNER ALEXANDRE LUCENA, Cenargen; MARIA CRISTINA MATTAR DA SILVA, Cenargen; OSMUNDO BRILHANTE OLIVEIRA‑NETO, Faculdade Planalto Central; MARIA FATIMA GROSSI DE SA, Cenargen. |
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Título: |
Improved cotton transformation protocol mediated by Agrobacterium and biolistic combined-methods. |
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Ano de publicação: |
2021 |
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Fonte/Imprenta: |
Planta, v. 254, 20, 2021. |
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ISSN: |
1432-2048; 0032-0935 |
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DOI: |
https://doi-org.ez103.periodicos.capes.gov.br/10.1007/s00425-021-03666-5 |
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Idioma: |
Inglês |
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Conteúdo: |
Cotton (Gossypium spp.) is the most important crop for natural textile fiber production worldwide. Nonetheless, one of the main challenges in cotton production are the losses resulting from insect pests, pathogens, and abiotic stresses. One effective way to solve these issues is to use genetically modified (GM) varieties. Herein, we describe an improved protocol for straightforward and cost-effective genetic transformation of cotton embryo axes, merging biolistics and Agrobacterium. The experimental steps include (1) Agrobacterium preparation, (2) seed sterilization, (3) cotton embryo excision, (4) lesion of shoot-cells by tungsten bombardment, (5) Agrobacterium-mediated transformation, (6) embryo co-culture, (7) regeneration and selection of transgenic plants in vitro, and (8) molecular characterization of plants. Due to the high regenerative power of the embryonic axis and the exceptional ability of the meristem cells for plant regeneration through organogenesis in vitro, this protocol can be performed in approximately 4?10 weeks, with an average plant regeneration of about 5.5% (±?0.53) and final average transformation efficiency of 60% (±?0.55). The transgene was stably inherited, and most transgenic plants hold a single copy of the transgene, as desirable and expected in Agrobacterium-mediated transformation. Additionally, the transgene was stably expressed over generations, and transgenic proteins could be detected at high levels in the T2 generation of GM cotton plants. The T2 progeny showed no phenotypic or productivity disparity compared to wild-type plants. Collectively, the use of cotton embryo axes and the enhanced DNA-delivery system by combining particle bombardment and Agrobacterium infection enabled efficient transgenic plant recovery, overcoming usual limitations associated with the recalcitrance of several cotton genotypes subjected to somatic embryogenesis. The improved approach states this method?s success for cotton genetic modification, allowing us to obtain GM cotton plants carrying traits, which are of fundamental relevance for the advancement of global agribusiness. MenosCotton (Gossypium spp.) is the most important crop for natural textile fiber production worldwide. Nonetheless, one of the main challenges in cotton production are the losses resulting from insect pests, pathogens, and abiotic stresses. One effective way to solve these issues is to use genetically modified (GM) varieties. Herein, we describe an improved protocol for straightforward and cost-effective genetic transformation of cotton embryo axes, merging biolistics and Agrobacterium. The experimental steps include (1) Agrobacterium preparation, (2) seed sterilization, (3) cotton embryo excision, (4) lesion of shoot-cells by tungsten bombardment, (5) Agrobacterium-mediated transformation, (6) embryo co-culture, (7) regeneration and selection of transgenic plants in vitro, and (8) molecular characterization of plants. Due to the high regenerative power of the embryonic axis and the exceptional ability of the meristem cells for plant regeneration through organogenesis in vitro, this protocol can be performed in approximately 4?10 weeks, with an average plant regeneration of about 5.5% (±?0.53) and final average transformation efficiency of 60% (±?0.55). The transgene was stably inherited, and most transgenic plants hold a single copy of the transgene, as desirable and expected in Agrobacterium-mediated transformation. Additionally, the transgene was stably expressed over generations, and transgenic proteins could be detected at high levels in the T2 generation of GM cotton plant... Mostrar Tudo |
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Palavras-Chave: |
Embryonic axis; Genotype-independent transformation; Modificação genética do algodão. |
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Thesagro: |
Agrobacterium Tumefaciens; Algodão; DNA; Gossypium Hirsutum; Método de Melhoramento; Planta Transgênica. |
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Thesaurus Nal: |
Abiotic stress; Biolistics; Cotton; Genetic transformation; Genetically modified plants. |
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Categoria do assunto: |
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G Melhoramento Genético |
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Marc: |
LEADER 03519naa a2200457 a 4500
001 2134441
005 2021-09-15
008 2021 bl uuuu u00u1 u #d
022 $a1432-2048; 0032-0935
024 7 $ahttps://doi-org.ez103.periodicos.capes.gov.br/10.1007/s00425-021-03666-5$2DOI
100 1 $aRIBEIRO, T. P.
245 $aImproved cotton transformation protocol mediated by Agrobacterium and biolistic combined-methods.$h[electronic resource]
260 $c2021
520 $aCotton (Gossypium spp.) is the most important crop for natural textile fiber production worldwide. Nonetheless, one of the main challenges in cotton production are the losses resulting from insect pests, pathogens, and abiotic stresses. One effective way to solve these issues is to use genetically modified (GM) varieties. Herein, we describe an improved protocol for straightforward and cost-effective genetic transformation of cotton embryo axes, merging biolistics and Agrobacterium. The experimental steps include (1) Agrobacterium preparation, (2) seed sterilization, (3) cotton embryo excision, (4) lesion of shoot-cells by tungsten bombardment, (5) Agrobacterium-mediated transformation, (6) embryo co-culture, (7) regeneration and selection of transgenic plants in vitro, and (8) molecular characterization of plants. Due to the high regenerative power of the embryonic axis and the exceptional ability of the meristem cells for plant regeneration through organogenesis in vitro, this protocol can be performed in approximately 4?10 weeks, with an average plant regeneration of about 5.5% (±?0.53) and final average transformation efficiency of 60% (±?0.55). The transgene was stably inherited, and most transgenic plants hold a single copy of the transgene, as desirable and expected in Agrobacterium-mediated transformation. Additionally, the transgene was stably expressed over generations, and transgenic proteins could be detected at high levels in the T2 generation of GM cotton plants. The T2 progeny showed no phenotypic or productivity disparity compared to wild-type plants. Collectively, the use of cotton embryo axes and the enhanced DNA-delivery system by combining particle bombardment and Agrobacterium infection enabled efficient transgenic plant recovery, overcoming usual limitations associated with the recalcitrance of several cotton genotypes subjected to somatic embryogenesis. The improved approach states this method?s success for cotton genetic modification, allowing us to obtain GM cotton plants carrying traits, which are of fundamental relevance for the advancement of global agribusiness.
650 $aAbiotic stress
650 $aBiolistics
650 $aCotton
650 $aGenetic transformation
650 $aGenetically modified plants
650 $aAgrobacterium Tumefaciens
650 $aAlgodão
650 $aDNA
650 $aGossypium Hirsutum
650 $aMétodo de Melhoramento
650 $aPlanta Transgênica
653 $aEmbryonic axis
653 $aGenotype-independent transformation
653 $aModificação genética do algodão
700 1 $aLOURENCO, I. T.
700 1 $aMELO, B. P. de
700 1 $aMORGANTE, C. V.
700 1 $aSALLES FILHO, A.
700 1 $aLINS, C. B. J.
700 1 $aFERREIRA, G. F.
700 1 $aMELLO, G. N.
700 1 $aMACEDO, L. L. P. de
700 1 $aLUCENA, W. A.
700 1 $aSILVA, M. C. M. da
700 1 $aOLIVEIRA‑NETO, O. B.
700 1 $aSA, M. F. G. de
773 $tPlanta$gv. 254, 20, 2021.
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Registro original: |
Embrapa Semiárido (CPATSA) |
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