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Quantitative transient GUS expression in J-104 rice calli through manipulation of in vitro culture conditions
Pérez Bernal, Maylin; Hernández, Carlos; Barceló, Teresa María; Delgado, Magalis; Armas, Raúl.
  • Pérez Bernal, Maylin; Center for Genetic Engineering and Biotechnology of Sancti Spiritus. Research Department. Sancti Spiritus. CU
  • Hernández, Carlos; Center for Genetic Engineering and Biotechnology of Sancti Spiritus. Research Department. Sancti Spiritus. CU
  • Barceló, Teresa María; Center for Genetic Engineering and Biotechnology of Sancti Spiritus. Research Department. Sancti Spiritus. CU
  • Delgado, Magalis; Laboratory Technician. Sancti Spiritus. CU
  • Armas, Raúl; Center for Genetic Engineering and Biotechnology of Sancti Spiritus. Research Department. Sancti Spiritus. CU
Rev. colomb. biotecnol ; 11(2): 75-84, dic. 2009.
Artigo em Inglês | LILACS | ID: lil-550522
ABSTRACT
This paper purposes suitable conditions for callus induction and co-cultivation with Agrobacterium tumefaciens of J-104 rice cultivar. It was evaluated the effect of different concentrations of 2.4-D and agar, and the inclusion of L-proline and L-glutamine in callus culture medium. The use of 2.5 mg/L 2.4-D and 0.8% agar allowed the highest percentage of embryogenic calli. Callus formation was improved considerably with 500 mg/L of L-proline and L-glutamine in the culture medium. Different factors were studied throughout co-cultivation of calli with A. tumefaciens inoculation time, co-cultivation temperature, concentration of acetosyringone and co-cultivation period. Transient GUS expression was quantified by fluorometry in all co-cultivated calli. The best results were obtained with the following conditions 10 min as inoculation time, 100µM acetosyringone in co-cultivation medium, temperature of 20ºC, and 3 days as co-cultivation period.
RESUMEN
Se describen las condiciones óptimas para la callogénesis y cocultivo de callos con Agrobacterium tume-faciens de la variedad de arroz J-104. Se determinó el efecto de diferentes concentraciones de 2.4-D, agar y de L-prolina y L-glutamina en el medio de cultivo de callos. El uso de 2,5 mg/L de 2.4-D y 0,8% de agar permitió lograr el porcentaje más alto de callos embriogénicos. La formación de callos fue mejorada considerablemente con la adición de 500 mg/L de L-prolina e igual concentración de L-glutamina en el medio de cultivo. Se estudiaron diferentes factores en el cocultivo de los callos con A. tumefaciens tiempo de inoculación, concentración de acetosiringona, temperatura y tiempo de cocultivo. Para comparar el efecto de cada factor sobre la expresión GUS se cuantificó la actividad transitoria mediante fluorimetría. Los valores más altos de actividad fluorimétrica fueron obtenidos con las siguientes condiciones 10 min de inoculación, 100µM de acetosiringona en el medio de cocultivo y 3 días de cocultivo a 20 ºC.
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Texto completo: DisponíveL Índice: LILACS (Américas) Assunto principal: Técnicas de Cocultura Idioma: Inglês Revista: Rev. colomb. biotecnol Assunto da revista: Biotecnologia Ano de publicação: 2009 Tipo de documento: Artigo País de afiliação: Cuba Instituição/País de afiliação: Center for Genetic Engineering and Biotechnology of Sancti Spiritus/CU / Laboratory Technician/CU

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Texto completo: DisponíveL Índice: LILACS (Américas) Assunto principal: Técnicas de Cocultura Idioma: Inglês Revista: Rev. colomb. biotecnol Assunto da revista: Biotecnologia Ano de publicação: 2009 Tipo de documento: Artigo País de afiliação: Cuba Instituição/País de afiliação: Center for Genetic Engineering and Biotechnology of Sancti Spiritus/CU / Laboratory Technician/CU