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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
Optimization of coffee (Coffea arabica) transformation parameters using uidA and hpt genes: effect of osmotic pre-treatment, helium pressure and target distance
dc.creator | Gatica Arias, Andrés Mauricio | |
dc.creator | Arrieta Espinoza, Griselda | |
dc.creator | Espinoza Esquivel, Ana Mercedes | |
dc.date | 2015-10-06 | |
dc.date.accessioned | 2016-05-03T15:25:38Z | |
dc.date.available | 2016-05-03T15:25:38Z | |
dc.identifier | http://revistas.ucr.ac.cr/index.php/rbt/article/view/21292 | |
dc.identifier | 10.15517/rbt.v57i0.21292 | |
dc.identifier.uri | https://hdl.handle.net/10669/25675 | |
dc.description | The 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.description | La 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.format | application/pdf | |
dc.language | eng | |
dc.publisher | Universidad de Costa Rica | en-US |
dc.rights | Copyright (c) 2015 International Journal of Tropical Biology and Conservation | en-US |
dc.source | Revista de Biología Tropical/International Journal of Tropical Biology and Conservation; Vol 57 (Suplemento 1) 2009; 151-160 | en-US |
dc.source | Revista de Biología Tropical/International Journal of Tropical Biology and Conservation; Vol 57 (Suplemento 1) 2009; 151-160 | es-ES |
dc.source | Revista Biología Tropical; Vol 57 (Suplemento 1) 2009; 151-160 | pt-PT |
dc.source | 2215-2075 | |
dc.source | 0034-7744 | |
dc.source | 10.15517/rbt.v57i0 | |
dc.subject | coffea arabica | en-US |
dc.subject | caturra | en-US |
dc.subject | catuaí | en-US |
dc.subject | biolistic delivery | en-US |
dc.subject | osmotic treatment | en-US |
dc.subject | uidA | en-US |
dc.subject | GUSPlus™ | en-US |
dc.subject | bar | en-US |
dc.subject | hpt | en-US |
dc.subject | nptII | en-US |
dc.subject | Costa Rica | en-US |
dc.subject | caturra | es-ES |
dc.subject | catuaí | es-ES |
dc.subject | biobalística | es-ES |
dc.subject | tratamiento osmótico | es-ES |
dc.subject | uidA | es-ES |
dc.subject | GUSPlus™ | es-ES |
dc.subject | bar | es-ES |
dc.subject | hpt | es-ES |
dc.subject | nptII | es-ES |
dc.subject | Costa Rica | es-ES |
dc.title | Optimization of coffee (Coffea arabica) transformation parameters using uidA and hpt genes: effect of osmotic pre-treatment, helium pressure and target distance | en-US |
dc.title | Optimization of coffee (Coffea arabica) transformation parameters using uidA and hpt genes: effect of osmotic pre-treatment, helium pressure and target distance | es-ES |
dc.type | artículo original |
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