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Registro Completo |
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Biblioteca(s): |
Embrapa Hortaliças; Embrapa Instrumentação. |
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Data corrente: |
18/11/2021 |
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Data da última atualização: |
23/01/2024 |
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Tipo da produção científica: |
Artigo em Periódico Indexado |
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Autoria: |
PETRÔNIO, M. S.; BARROS-ALEXANDRINO, T. T.; LIMA, A. M. F.; ASSIS, O. B. G. de; INOUE-NAGATA, A. K.; NAKASU, E. Y. T.; TIERA, M. J.; PILON, L. |
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Afiliação: |
ODILIO BENEDITO GARRIDO DE ASSIS, CNPDIA; ALICE KAZUKO INOUE NAGATA, CNPH; ERICH YUKIO TEMPEL NAKASU, CNPH; LUCIMEIRE PILON, CNPH. |
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Título: |
Physicochemical and toxicity investigation of chitosan-based dsRNA nanocarrier formation. |
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Ano de publicação: |
2022 |
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Fonte/Imprenta: |
Biointerface Research in Applied Chemistry, vol. 12, n. 4, 2022. |
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Páginas: |
5266 - 5279 |
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DOI: |
https://doi.org/10.33263/BRIAC124.52665279 |
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Idioma: |
Inglês |
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Conteúdo: |
Technologies involving the use of double-stranded RNA (dsRNA) to elicit RNA interference (RNAi) in pest control have emerged as an alternative to traditional pesticides. RNAi can mediate natural cell protection being a promising tool to provide prompt responses in plant defense against pathogens. The present study is focused on the physicochemical characterization of formed dsRNAloaded nanoparticles as a result of chitosan-dsRNA ionic interactions. Additionally, a preliminary investigation was conducted of the in-vitro toxicity of loaded nanoparticles in lettuce and human red blood cells. dsRNA molecules, homologous to partial phytopathogenic tomato mosaic virus (ToMV) sequence, were used as a model. The main groups involved in the chitosan-dsRNA ionic coupling were identified by Fourier-transform infrared spectroscopy, and the stability of formed nanoparticles was accessed by dynamic light scattering, electrophoresis, and thermal analyses. The chitosan showed a higher ability to bind to dsRNA at low charge ratios (N/P = 1), ruled by positively charged chitosan methyl groups and negatively charged phosphate groups from the RNA backbone, resulting in small nanoparticles (73.25 nm size) at low polydispersity (0.25). The toxic assays of these particles, on lettuce seeds and in human erythrocytes, revealed very low toxicity demonstrating their safety as a platform, thereby holding potential use as biodefensive for crop protection. |
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Palavras-Chave: |
Chitosan nanocarrier; DsRNA phytotoxicity assay; Tomato mosaic virus (ToMV). |
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Thesaurus Nal: |
Double-stranded RNA; Lettuce; RNA interference; Tomato mosaic virus. |
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Categoria do assunto: |
-- |
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URL: |
https://ainfo.cnptia.embrapa.br/digital/bitstream/item/232647/1/P-Physicochemical-and-Toxicity-Investigation-of-Chitosanbased-dsRNA-Nanocarrier-Formation.pdf
https://ainfo.cnptia.embrapa.br/digital/bitstream/item/227886/1/20695837124.526652791.pdf
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Marc: |
LEADER 02428naa a2200313 a 4500
001 2141015
005 2024-01-23
008 2022 bl uuuu u00u1 u #d
024 7 $ahttps://doi.org/10.33263/BRIAC124.52665279$2DOI
100 1 $aPETRÔNIO, M. S.
245 $aPhysicochemical and toxicity investigation of chitosan-based dsRNA nanocarrier formation.$h[electronic resource]
260 $c2022
300 $a5266 - 5279
520 $aTechnologies involving the use of double-stranded RNA (dsRNA) to elicit RNA interference (RNAi) in pest control have emerged as an alternative to traditional pesticides. RNAi can mediate natural cell protection being a promising tool to provide prompt responses in plant defense against pathogens. The present study is focused on the physicochemical characterization of formed dsRNAloaded nanoparticles as a result of chitosan-dsRNA ionic interactions. Additionally, a preliminary investigation was conducted of the in-vitro toxicity of loaded nanoparticles in lettuce and human red blood cells. dsRNA molecules, homologous to partial phytopathogenic tomato mosaic virus (ToMV) sequence, were used as a model. The main groups involved in the chitosan-dsRNA ionic coupling were identified by Fourier-transform infrared spectroscopy, and the stability of formed nanoparticles was accessed by dynamic light scattering, electrophoresis, and thermal analyses. The chitosan showed a higher ability to bind to dsRNA at low charge ratios (N/P = 1), ruled by positively charged chitosan methyl groups and negatively charged phosphate groups from the RNA backbone, resulting in small nanoparticles (73.25 nm size) at low polydispersity (0.25). The toxic assays of these particles, on lettuce seeds and in human erythrocytes, revealed very low toxicity demonstrating their safety as a platform, thereby holding potential use as biodefensive for crop protection.
650 $aDouble-stranded RNA
650 $aLettuce
650 $aRNA interference
650 $aTomato mosaic virus
653 $aChitosan nanocarrier
653 $aDsRNA phytotoxicity assay
653 $aTomato mosaic virus (ToMV)
700 1 $aBARROS-ALEXANDRINO, T. T.
700 1 $aLIMA, A. M. F.
700 1 $aASSIS, O. B. G. de
700 1 $aINOUE-NAGATA, A. K.
700 1 $aNAKASU, E. Y. T.
700 1 $aTIERA, M. J.
700 1 $aPILON, L.
773 $tBiointerface Research in Applied Chemistry, vol. 12$gn. 4, 2022.
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Registro original: |
Embrapa Instrumentação (CNPDIA) |
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 | Acesso ao texto completo restrito à biblioteca da Embrapa Arroz e Feijão. Para informações adicionais entre em contato com cnpaf.biblioteca@embrapa.br. |
Registro Completo
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Biblioteca(s): |
Embrapa Arroz e Feijão; Embrapa Recursos Genéticos e Biotecnologia. |
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Data corrente: |
09/08/2013 |
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Data da última atualização: |
08/03/2023 |
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Tipo da produção científica: |
Artigo em Periódico Indexado |
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Circulação/Nível: |
A - 1 |
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Autoria: |
ARAGÃO, F. J. L.; NOGUEIRA, E. O. P. L.; TINOCO, M. L. P.; FARIA, J. C. |
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Afiliação: |
FRANCISCO JOSE LIMA ARAGAO, CENARGEN; ELSA OLIVEIRA P E LAGO NOGUEIRA, CENARGEN; MARIA LAINE P. TINOCO, CENARGEN; JOSIAS CORREA DE FARIA, CNPAF. |
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Título: |
Molecular characterization of the first commercial transgenic common bean immune to the Bean golden mosaic virus. |
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Ano de publicação: |
2013 |
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Fonte/Imprenta: |
Journal of Biotechnology, Amsterdam, v. 66, n. 1/2, p. 42-50, June 2013. |
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DOI: |
http://dx.doi.org/10.1016/j.jbiotec.2013.04.009 |
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Idioma: |
Inglês |
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Conteúdo: |
Golden mosaic of common bean is caused by the Bean golden mosaic virus (BGMV). The disease is one of the greatest constraints on bean production in Latin America and causes significant yield losses. The RNAi concept was explored to silence the rep (AC1) viral gene and a transgenic bean line immune to BGMV upon inoculation at high pressure was previously generated. Identification of the transgene insert confirmed the presence of a single locus corresponding to two intact copies of the RNAi cassette in opposite orientation and three intact copies of the AtAhas gene. It is flanked by Phaseolus genomic sequences and interspersed by one nuclear and three chloroplastic genomic sequences. Southern analyses showed that the transgenes were structurally stable for eight self-pollinated generations and after backcrosses with a non transgenic commercial variety. Transgene expression analyses revealed similar levels of siRNA in leaves of transgenic plants cultivated under field conditions in three distinct regions. siRNA were also analyzed during seed development in common bean transgenic plants. siRNA signals were also detected in seeds, albeit at significantly lower levels than those observed in leaves, and could not be detected in seeds cooked during 10 min. This information is relevant to demonstrate that GM beans are free of siRNA signals after cooking and therefore suitable for human consumption. Additionally, characterization of the locus where the transgene was integrated in the common bean genome provides a valuable tool to trace this GM bean material in the field and in the market. MenosGolden mosaic of common bean is caused by the Bean golden mosaic virus (BGMV). The disease is one of the greatest constraints on bean production in Latin America and causes significant yield losses. The RNAi concept was explored to silence the rep (AC1) viral gene and a transgenic bean line immune to BGMV upon inoculation at high pressure was previously generated. Identification of the transgene insert confirmed the presence of a single locus corresponding to two intact copies of the RNAi cassette in opposite orientation and three intact copies of the AtAhas gene. It is flanked by Phaseolus genomic sequences and interspersed by one nuclear and three chloroplastic genomic sequences. Southern analyses showed that the transgenes were structurally stable for eight self-pollinated generations and after backcrosses with a non transgenic commercial variety. Transgene expression analyses revealed similar levels of siRNA in leaves of transgenic plants cultivated under field conditions in three distinct regions. siRNA were also analyzed during seed development in common bean transgenic plants. siRNA signals were also detected in seeds, albeit at significantly lower levels than those observed in leaves, and could not be detected in seeds cooked during 10 min. This information is relevant to demonstrate that GM beans are free of siRNA signals after cooking and therefore suitable for human consumption. Additionally, characterization of the locus where the transgene was integrated in the ... Mostrar Tudo |
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Thesagro: |
Engenharia genética; Feijão. |
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Thesaurus NAL: |
Begomovirus; Dry beans; Geminiviridae; Genetic engineering; RNA interference. |
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Categoria do assunto: |
X Pesquisa, Tecnologia e Engenharia |
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Marc: |
LEADER 02424naa a2200253 a 4500
001 1968340
005 2023-03-08
008 2013 bl uuuu u00u1 u #d
024 7 $ahttp://dx.doi.org/10.1016/j.jbiotec.2013.04.009$2DOI
100 1 $aARAGÃO, F. J. L.
245 $aMolecular characterization of the first commercial transgenic common bean immune to the Bean golden mosaic virus.$h[electronic resource]
260 $c2013
520 $aGolden mosaic of common bean is caused by the Bean golden mosaic virus (BGMV). The disease is one of the greatest constraints on bean production in Latin America and causes significant yield losses. The RNAi concept was explored to silence the rep (AC1) viral gene and a transgenic bean line immune to BGMV upon inoculation at high pressure was previously generated. Identification of the transgene insert confirmed the presence of a single locus corresponding to two intact copies of the RNAi cassette in opposite orientation and three intact copies of the AtAhas gene. It is flanked by Phaseolus genomic sequences and interspersed by one nuclear and three chloroplastic genomic sequences. Southern analyses showed that the transgenes were structurally stable for eight self-pollinated generations and after backcrosses with a non transgenic commercial variety. Transgene expression analyses revealed similar levels of siRNA in leaves of transgenic plants cultivated under field conditions in three distinct regions. siRNA were also analyzed during seed development in common bean transgenic plants. siRNA signals were also detected in seeds, albeit at significantly lower levels than those observed in leaves, and could not be detected in seeds cooked during 10 min. This information is relevant to demonstrate that GM beans are free of siRNA signals after cooking and therefore suitable for human consumption. Additionally, characterization of the locus where the transgene was integrated in the common bean genome provides a valuable tool to trace this GM bean material in the field and in the market.
650 $aBegomovirus
650 $aDry beans
650 $aGeminiviridae
650 $aGenetic engineering
650 $aRNA interference
650 $aEngenharia genética
650 $aFeijão
700 1 $aNOGUEIRA, E. O. P. L.
700 1 $aTINOCO, M. L. P.
700 1 $aFARIA, J. C.
773 $tJournal of Biotechnology, Amsterdam$gv. 66, n. 1/2, p. 42-50, June 2013.
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Embrapa Recursos Genéticos e Biotecnologia (CENARGEN) |
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