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Optimization of coffee (Coffea arabica) transformation parameters using uidA and hpt genes: effect of osmotic pre-treatment, helium pressure and target distance

dc.creatorGatica Arias, Andrés Mauricio
dc.creatorArrieta Espinoza, Griselda
dc.creatorEspinoza Esquivel, Ana Mercedes
dc.date2015-10-06
dc.date.accessioned2016-05-03T15:25:38Z
dc.date.available2016-05-03T15:25:38Z
dc.identifierhttp://revistas.ucr.ac.cr/index.php/rbt/article/view/21292
dc.identifier10.15517/rbt.v57i0.21292
dc.identifier.urihttps://hdl.handle.net/10669/25675
dc.descriptionThe aim of this work was to optimize the biolistic delivery parameters that affect the DNA delivery and stable expression of marker genes into coffee tissues (Coffea arabica. L. cvs. Caturra and Catuaí). The effect of osmotic preculture length, osmotic concentration of medium, Helium pressure and target distance on transient expression of the uidA gene in coffee leaves and somatic embryos were tested. The highest transient uidA expression was obtained when Caturra (18.3±2.8) and Catuaí (6.8±2.0) leaves and Catuaí embryos (80.0±7.4) were cultured for 5h on Yasuda medium complemented with 0.5M Mannitol +0.5M Sorbitol. The combination of 1100psi and a target distance of 9cm resulted in the highest number of blue spots per Caturra leaf segment (23.6±3.9), whereas for the Catuaí variety the combination of 1100psi and a target distance of six (10.2±1.9) and nine (8.2±1.9) cm gave the highest number of blue spots per leaf segment. The optimized protocol was tested with pCAMBIA 1 301 (uidA gene and the hpt gene), pCAMBIA 1 305.2 (uidA version GUSPlus ™ and the hpt gene) and pCAMBIA 1 301-BAR (uidA gene and the bar gene). The highest number of blue spots was obtained when Caturra (54.6±5.7) and Catuaí (28.9±4.3) leaves were bombarded with pCAMBIA 1 305.2. Selection of bombarded coffee tissues with 100mg/l hygromicyn caused the oxidation of tissues.en-US
dc.descriptionLa presente investigación tuvo como objetivo optimizar los parámetros que afectan la incorporación y expresión de genes marcadores mediante biobalística en segmentos de hoja y embriones somáticos de café (Coffea arabica. L. cvs. Caturra y Catuaí). La mayor expresión transitoria del gen uidA en segmentos de hoja de Caturra (18.3±2.8) y Catuaí (6.8±2.0) y embriones somáticos de Catuaí (80.0±7.4) se obtuvo al cultivar los explantes por cinco horas previo al bombardeo en el medio Yasuda complementado con 0.5M mannitol+0.5M sorbitol. Asimismo, se obtuvo una mayor expresión transitoria del gen uidA al bombardear los segmentos de hoja de Caturra y Catuaí y embriones somáticos de Catuaí con una presión de helio de 1 100psi y una distancia de bombardeo de 6 o 9 cm.es-ES
dc.formatapplication/pdf
dc.languageeng
dc.publisherUniversidad de Costa Ricaen-US
dc.rightsCopyright (c) 2015 International Journal of Tropical Biology and Conservationen-US
dc.sourceRevista de Biología Tropical/International Journal of Tropical Biology and Conservation; Vol 57 (Suplemento 1) 2009; 151-160en-US
dc.sourceRevista de Biología Tropical/International Journal of Tropical Biology and Conservation; Vol 57 (Suplemento 1) 2009; 151-160es-ES
dc.sourceRevista Biología Tropical; Vol 57 (Suplemento 1) 2009; 151-160pt-PT
dc.source2215-2075
dc.source0034-7744
dc.source10.15517/rbt.v57i0
dc.subjectcoffea arabicaen-US
dc.subjectcaturraen-US
dc.subjectcatuaíen-US
dc.subjectbiolistic deliveryen-US
dc.subjectosmotic treatmenten-US
dc.subjectuidAen-US
dc.subjectGUSPlus™en-US
dc.subjectbaren-US
dc.subjecthpten-US
dc.subjectnptIIen-US
dc.subjectCosta Ricaen-US
dc.subjectcaturraes-ES
dc.subjectcatuaíes-ES
dc.subjectbiobalísticaes-ES
dc.subjecttratamiento osmóticoes-ES
dc.subjectuidAes-ES
dc.subjectGUSPlus™es-ES
dc.subjectbares-ES
dc.subjecthptes-ES
dc.subjectnptIIes-ES
dc.subjectCosta Ricaes-ES
dc.titleOptimization of coffee (Coffea arabica) transformation parameters using uidA and hpt genes: effect of osmotic pre-treatment, helium pressure and target distanceen-US
dc.titleOptimization of coffee (Coffea arabica) transformation parameters using uidA and hpt genes: effect of osmotic pre-treatment, helium pressure and target distancees-ES
dc.typeartículo original


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