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Registro Completo |
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Biblioteca(s): |
Embrapa Café. |
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Data corrente: |
16/11/2022 |
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Data da última atualização: |
16/11/2022 |
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Tipo da produção científica: |
Artigo em Periódico Indexado |
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Autoria: |
CASARIN, T.; FREITAS, N. C.; PINTO, R. T.; BREITLER, J. C.; RODRIGUES, L. A. Z.; MARRACCINI, P.; ETIENNE, H.; DINIZ, L. E. C.; ANDRADE, A. C.; PAIVA, L. V. |
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Afiliação: |
TATIANE CASARIN, UNIVERSIDADE FEDERAL DE LAVRAS; NATÁLIA CHAGAS FREITAS, UNIVERSIDADE FEDERAL DE LAVRAS; RENAN TERASSI PINTO, UNIVERSIDADE FEDERAL DE LAVRAS; JEAN‐CHRISTOPHE BREITLER, CENTRO DE COOPERAÇÃO INTERNACIONAL EM PESQUISA AGRÍCOLA PARA O DESENVOLVIMENTO; LEONARDO AUGUSTO ZEBRAL RODRIGUES, UNIVERSIDADE FEDERAL DE LAVRAS; PIERRE MARRACCINI, CENTRO DE COOPERAÇÃO INTERNACIONAL EM PESQUISA AGRÍCOLA PARA O DESENVOLVIMENTO; HERVÉ ETIENNE, CENTRO DE COOPERAÇÃO INTERNACIONAL EM PESQUISA AGRÍCOLA PARA O DESENVOLVIMENTO; LEANDRO EUGENIO CARDAMONE DINIZ, CNPSO; ALAN CARVALHO ANDRADE, CNPCa; LUCIANO VILELA PAIVA, UNIVERSIDADE FEDERAL DE LAVRAS. |
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Título: |
Multiplex CRISPR/Cas9-mediated knockout of the phytoene desaturase gene in Coffea canéfora. |
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Ano de publicação: |
2022 |
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Fonte/Imprenta: |
Scientific Reports, v. 12, 17270, 2022. 10 p. |
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DOI: |
https://doi.org/10.1038/s41598-022-21566-w |
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Idioma: |
Inglês |
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Conteúdo: |
Coffea canephora (2n=2x22 chromosomes) is a species with extensive genetic diversity and desirable agronomic traits for coffee breeding programs. However, obtaining a new coffee cultivar through conventional breeding techniques may require more than 30 years of crossing cycles and selection, which hampers the effort of keeping up with market demands and rapidly proposing more resilient to climate change varieties. Although, the application of modern biotechnology tools such as precision genetic engineering technologies may enable a faster cultivar development process. Therefore, we aimed to validate the CRISPR/Cas9 system to generate mutations on a selected genotype of C. canephora, the clone 14. Embryogenic calli and a multiplex binary vector containing two sgRNAs targeting different exons of the CcPDS gene were used. The sgRNAs were under the C. canephora U6 promoter regulation. The target gene encodes phytoene desaturase, an enzyme essential for photosynthesis involved in B-carotene biosynthesis. Somatic seedlings and embryos with albino, variegated and green phenotypes regenerated after Agrobacterium tumefaciens-mediated genetic transformation were analyzed by verifying the insertion of the Cas9 gene and later by sequencing the sgRNAs target regions in the genome of Robusta modified seedlings. Among them, 77% had the expected mutations, and of which, 50% of them had at least one target with a homozygous mutation. The genotype, temperature of co-cultivation with the bacteria, and light intensity used for subsequent embryo regeneration appeared to strongly influence the successful regeneration of plants with a mutated CcPDS gene in the Coffea genus. MenosCoffea canephora (2n=2x22 chromosomes) is a species with extensive genetic diversity and desirable agronomic traits for coffee breeding programs. However, obtaining a new coffee cultivar through conventional breeding techniques may require more than 30 years of crossing cycles and selection, which hampers the effort of keeping up with market demands and rapidly proposing more resilient to climate change varieties. Although, the application of modern biotechnology tools such as precision genetic engineering technologies may enable a faster cultivar development process. Therefore, we aimed to validate the CRISPR/Cas9 system to generate mutations on a selected genotype of C. canephora, the clone 14. Embryogenic calli and a multiplex binary vector containing two sgRNAs targeting different exons of the CcPDS gene were used. The sgRNAs were under the C. canephora U6 promoter regulation. The target gene encodes phytoene desaturase, an enzyme essential for photosynthesis involved in B-carotene biosynthesis. Somatic seedlings and embryos with albino, variegated and green phenotypes regenerated after Agrobacterium tumefaciens-mediated genetic transformation were analyzed by verifying the insertion of the Cas9 gene and later by sequencing the sgRNAs target regions in the genome of Robusta modified seedlings. Among them, 77% had the expected mutations, and of which, 50% of them had at least one target with a homozygous mutation. The genotype, temperature of co-cultivation with the bacte... Mostrar Tudo |
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Thesagro: |
Biodiversidade; Café Robusta; Coffea Canephora; Genótipo; Melhoramento Genético Vegetal; Mutação. |
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Categoria do assunto: |
-- |
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URL: |
https://ainfo.cnptia.embrapa.br/digital/bitstream/doc/1148303/1/Multiplex-CRISPRCas9-mediated.pdf
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Marc: |
LEADER 02597naa a2200313 a 4500
001 2148303
005 2022-11-16
008 2022 bl uuuu u00u1 u #d
024 7 $ahttps://doi.org/10.1038/s41598-022-21566-w$2DOI
100 1 $aCASARIN, T.
245 $aMultiplex CRISPR/Cas9-mediated knockout of the phytoene desaturase gene in Coffea canéfora.$h[electronic resource]
260 $c2022
520 $aCoffea canephora (2n=2x22 chromosomes) is a species with extensive genetic diversity and desirable agronomic traits for coffee breeding programs. However, obtaining a new coffee cultivar through conventional breeding techniques may require more than 30 years of crossing cycles and selection, which hampers the effort of keeping up with market demands and rapidly proposing more resilient to climate change varieties. Although, the application of modern biotechnology tools such as precision genetic engineering technologies may enable a faster cultivar development process. Therefore, we aimed to validate the CRISPR/Cas9 system to generate mutations on a selected genotype of C. canephora, the clone 14. Embryogenic calli and a multiplex binary vector containing two sgRNAs targeting different exons of the CcPDS gene were used. The sgRNAs were under the C. canephora U6 promoter regulation. The target gene encodes phytoene desaturase, an enzyme essential for photosynthesis involved in B-carotene biosynthesis. Somatic seedlings and embryos with albino, variegated and green phenotypes regenerated after Agrobacterium tumefaciens-mediated genetic transformation were analyzed by verifying the insertion of the Cas9 gene and later by sequencing the sgRNAs target regions in the genome of Robusta modified seedlings. Among them, 77% had the expected mutations, and of which, 50% of them had at least one target with a homozygous mutation. The genotype, temperature of co-cultivation with the bacteria, and light intensity used for subsequent embryo regeneration appeared to strongly influence the successful regeneration of plants with a mutated CcPDS gene in the Coffea genus.
650 $aBiodiversidade
650 $aCafé Robusta
650 $aCoffea Canephora
650 $aGenótipo
650 $aMelhoramento Genético Vegetal
650 $aMutação
700 1 $aFREITAS, N. C.
700 1 $aPINTO, R. T.
700 1 $aBREITLER, J. C.
700 1 $aRODRIGUES, L. A. Z.
700 1 $aMARRACCINI, P.
700 1 $aETIENNE, H.
700 1 $aDINIZ, L. E. C.
700 1 $aANDRADE, A. C.
700 1 $aPAIVA, L. V.
773 $tScientific Reports$gv. 12, 17270, 2022. 10 p.
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Registro original: |
Embrapa Café (CNPCa) |
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 | Acesso ao texto completo restrito à biblioteca da Embrapa Solos. Para informações adicionais entre em contato com cnps.biblioteca@embrapa.br. |
Registro Completo
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Biblioteca(s): |
Embrapa Solos. |
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Data corrente: |
02/06/2014 |
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Data da última atualização: |
21/06/2021 |
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Autoria: |
CAMARGO, M. N.; JACOMINE, P. K. T.; CARVALHO, A. P. de; LARACH, J. O. I. |
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Afiliação: |
MARCELO NUNES CAMARGO, CNPS; PAULO KLINGER TITO JACOMINE, CNPS; AMÉRICO PEREIRA DE CARVALHO, SNLCS; JORGE OLMOS ITURRI LARACH, CNPS. |
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Título: |
Ensaio de estruturação de classes dos níveis hierárquicos de categorias II III IV, referentes à classe 8 (solos com B latossólico) do nível de categoria I. |
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Ano de publicação: |
1984 |
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Fonte/Imprenta: |
Rio de Janeiro: EMBRAPA-SNLCS, 1984. |
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Páginas: |
15 p. |
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Idioma: |
Português |
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Notas: |
Segundo estudo preparatório para futura 3ª aproximação do desenvolvimento do sistema. |
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Palavras-Chave: |
Niveis hierárquicos; SiBCS. |
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Thesagro: |
Classificação do Solo. |
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Categoria do assunto: |
P Recursos Naturais, Ciências Ambientais e da Terra |
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Marc: |
LEADER 00714nam a2200193 a 4500
001 1987431
005 2021-06-21
008 1984 bl uuuu u0uu1 u #d
100 1 $aCAMARGO, M. N.
245 $aEnsaio de estruturação de classes dos níveis hierárquicos de categorias II III IV, referentes à classe 8 (solos com B latossólico) do nível de categoria I.
260 $aRio de Janeiro: EMBRAPA-SNLCS$c1984
300 $a15 p.
500 $aSegundo estudo preparatório para futura 3ª aproximação do desenvolvimento do sistema.
650 $aClassificação do Solo
653 $aNiveis hierárquicos
653 $aSiBCS
700 1 $aJACOMINE, P. K. T.
700 1 $aCARVALHO, A. P. de
700 1 $aLARACH, J. O. I.
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