Registro Completo |
Biblioteca(s): |
Embrapa Recursos Genéticos e Biotecnologia; Embrapa Semiárido. |
Data corrente: |
18/09/2020 |
Data da última atualização: |
17/11/2020 |
Tipo da produção científica: |
Artigo em Periódico Indexado |
Autoria: |
MELO, B. P. de; LOURENCO-TESSUTTI, I. T.; MORGANTE, C. V.; SANTOS, N. C.; PINHEIRO, L. B.; LINS, C. B. de J.; SILVA, M. C. M.; MACEDO, L. L. P.; FONTES, E. P. B.; GROSSI-DE-SA, M. F. |
Afiliação: |
BRUNO PAES DE MELO, UFV; ISABELA TRISTAN LOURENCO TESSUTTI, Cenargen; CAROLINA VIANNA MORGANTE, CPATSA; NAIARA CORDEIRO SANTOS; LUANNA BEZERRA PINHEIRO, UCB; CAMILA BARROZO DE JESUS LINS; MARIA CRISTINA MATTAR DA SILVA, Cenargen; LEONARDO LIMA PEPINO DE MACEDO, Cenargen; ELIZABETH PACHECO BATISTA FONTES, UFV; MARIA FATIMA GROSSI DE SA, Cenargen. |
Título: |
Soybean embryonic axis transformation: combining biolistic and Agrobacterium-Mediated Protocols to overcome typical complications of in vitro plant regeneration. |
Ano de publicação: |
2020 |
Fonte/Imprenta: |
Frontiers in Plant Science, v. 11, article 1228, 2020. |
DOI: |
https://doi.org/10.3389/fpls.2020.01228 |
Idioma: |
Inglês |
Conteúdo: |
The first successful attempt to generate genetically modified plants expressing a transgene was preformed via T-DNA-based gene transfer employing Agrobacterium tumefaciens-mediated genetic transformation. Limitations over infectivity and in vitro tissue culture led to the development of other DNA delivery systems, such as the biolistic method. Herein, we developed a new one-step protocol for transgenic soybean recovery by combining the two different transformation methods. This protocol comprises the following steps: agrobacterial preparation, seed sterilization, soybean embryo excision, shoot-cell injury by tungsten-microparticle bombardment, A. tumefaciens-mediated transformation, embryo co-cultivation in vitro, and selection of transgenic plants. This protocol can be completed in approximately 30?40 weeks. The average efficiency of producing transgenic soybean germlines using this protocol was 9.84%, similar to other previously described protocols. However, we introduced a more cost-effective, more straightforward and shorter methodology for transgenic plant recovery, which allows co-cultivation and plant regeneration in a single step, decreasing the chances of contamination and making the manipulation easier. Finally, as a hallmark, our protocol does not generate plant chimeras, in contrast to traditional plant regeneration protocols applied in other Agrobacterium-mediated transformation methods. Therefore, this new approach of plant transformation is applicable for studies of gene function and the production of transgenic cultivars carrying different traits for precision-breeding programs. MenosThe first successful attempt to generate genetically modified plants expressing a transgene was preformed via T-DNA-based gene transfer employing Agrobacterium tumefaciens-mediated genetic transformation. Limitations over infectivity and in vitro tissue culture led to the development of other DNA delivery systems, such as the biolistic method. Herein, we developed a new one-step protocol for transgenic soybean recovery by combining the two different transformation methods. This protocol comprises the following steps: agrobacterial preparation, seed sterilization, soybean embryo excision, shoot-cell injury by tungsten-microparticle bombardment, A. tumefaciens-mediated transformation, embryo co-cultivation in vitro, and selection of transgenic plants. This protocol can be completed in approximately 30?40 weeks. The average efficiency of producing transgenic soybean germlines using this protocol was 9.84%, similar to other previously described protocols. However, we introduced a more cost-effective, more straightforward and shorter methodology for transgenic plant recovery, which allows co-cultivation and plant regeneration in a single step, decreasing the chances of contamination and making the manipulation easier. Finally, as a hallmark, our protocol does not generate plant chimeras, in contrast to traditional plant regeneration protocols applied in other Agrobacterium-mediated transformation methods. Therefore, this new approach of plant transformation is applicable for stud... Mostrar Tudo |
Palavras-Chave: |
Agrobacterium-mediated transformation; Embryonic axis; High-efficiency plant transformation; Particle bombardment; Planta geneticamente modificada; Recuperação trangênica de soja. |
Thesagro: |
Cultura de Tecido; Glycine Max; Soja. |
Thesaurus Nal: |
Agrobacterium; Embryonic structures; Genetic transformation; Plant genetics. |
Categoria do assunto: |
-- G Melhoramento Genético |
URL: |
https://ainfo.cnptia.embrapa.br/digital/bitstream/item/216085/1/fpls-11-01228.pdf
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Marc: |
LEADER 02941naa a2200397 a 4500 001 2125013 005 2020-11-17 008 2020 bl uuuu u00u1 u #d 024 7 $ahttps://doi.org/10.3389/fpls.2020.01228$2DOI 100 1 $aMELO, B. P. de 245 $aSoybean embryonic axis transformation$bcombining biolistic and Agrobacterium-Mediated Protocols to overcome typical complications of in vitro plant regeneration.$h[electronic resource] 260 $c2020 520 $aThe first successful attempt to generate genetically modified plants expressing a transgene was preformed via T-DNA-based gene transfer employing Agrobacterium tumefaciens-mediated genetic transformation. Limitations over infectivity and in vitro tissue culture led to the development of other DNA delivery systems, such as the biolistic method. Herein, we developed a new one-step protocol for transgenic soybean recovery by combining the two different transformation methods. This protocol comprises the following steps: agrobacterial preparation, seed sterilization, soybean embryo excision, shoot-cell injury by tungsten-microparticle bombardment, A. tumefaciens-mediated transformation, embryo co-cultivation in vitro, and selection of transgenic plants. This protocol can be completed in approximately 30?40 weeks. The average efficiency of producing transgenic soybean germlines using this protocol was 9.84%, similar to other previously described protocols. However, we introduced a more cost-effective, more straightforward and shorter methodology for transgenic plant recovery, which allows co-cultivation and plant regeneration in a single step, decreasing the chances of contamination and making the manipulation easier. Finally, as a hallmark, our protocol does not generate plant chimeras, in contrast to traditional plant regeneration protocols applied in other Agrobacterium-mediated transformation methods. Therefore, this new approach of plant transformation is applicable for studies of gene function and the production of transgenic cultivars carrying different traits for precision-breeding programs. 650 $aAgrobacterium 650 $aEmbryonic structures 650 $aGenetic transformation 650 $aPlant genetics 650 $aCultura de Tecido 650 $aGlycine Max 650 $aSoja 653 $aAgrobacterium-mediated transformation 653 $aEmbryonic axis 653 $aHigh-efficiency plant transformation 653 $aParticle bombardment 653 $aPlanta geneticamente modificada 653 $aRecuperação trangênica de soja 700 1 $aLOURENCO-TESSUTTI, I. T. 700 1 $aMORGANTE, C. V. 700 1 $aSANTOS, N. C. 700 1 $aPINHEIRO, L. B. 700 1 $aLINS, C. B. de J. 700 1 $aSILVA, M. C. M. 700 1 $aMACEDO, L. L. P. 700 1 $aFONTES, E. P. B. 700 1 $aGROSSI-DE-SA, M. F. 773 $tFrontiers in Plant Science$gv. 11, article 1228, 2020.
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Registro original: |
Embrapa Recursos Genéticos e Biotecnologia (CENARGEN) |
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