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

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 segme...

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Autores principales:	Gatica, Andrés M, Arrieta, Griselda, Espinoza, Ana M
Formato:	Online
Idioma:	eng
Publicado:	Universidad de Costa Rica 2009
Acceso en línea:	https://revistas.ucr.ac.cr/index.php/rbt/article/view/21292

id	RBT21292
record_format	ojs
spelling	RBT212922022-06-06T18:29:01Z 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 Gatica, Andrés M Arrieta, Griselda Espinoza, Ana M coffea arabica caturra catuaí biolistic delivery osmotic treatment uidA GUSPlus™ bar hpt nptII Costa Rica caturra catuaí biobalística tratamiento osmótico uidA GUSPlus™ bar hpt nptII Costa Rica 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. 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. Universidad de Costa Rica 2009-11-01 info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion Article application/pdf https://revistas.ucr.ac.cr/index.php/rbt/article/view/21292 10.15517/rbt.v57i0.21292 Revista de Biología Tropical; Vol. 57 No. S1 (2009): Volume 57 – Supplement 1 – November 2009: 50th anniversary School of Biology, University of Costa Rica; 151–160 Revista de Biología Tropical; Vol. 57 Núm. S1 (2009): Volumen 57 – Suplemento 1 – Noviembre: 50 aniversario Escuela de Biología, Universidad de Costa; 151–160 Revista Biología Tropical; Vol. 57 N.º S1 (2009): Volume 57 – Supplement 1 – November 2009: 50th anniversary School of Biology, University of Costa Rica; 151–160 2215-2075 0034-7744 10.15517/rbt.v57i0 eng https://revistas.ucr.ac.cr/index.php/rbt/article/view/21292/21506 Copyright (c) 2009 Revista de Biología Tropical http://creativecommons.org/licenses/by/4.0
institution	Universidad de Costa Rica
collection	Revista de Biología Tropical
language	eng
format	Online
author	Gatica, Andrés M Arrieta, Griselda Espinoza, Ana M
spellingShingle	Gatica, Andrés M Arrieta, Griselda Espinoza, Ana M Optimization of coffee (Coffea arabica) transformation parameters using uidA and hpt genes: effect of osmotic pre-treatment, helium pressure and target distance
author_facet	Gatica, Andrés M Arrieta, Griselda Espinoza, Ana M
author_sort	Gatica, Andrés M
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.
title	Optimization of coffee (Coffea arabica) transformation parameters using uidA and hpt genes: effect of osmotic pre-treatment, helium pressure and target distance
title_short	Optimization of coffee (Coffea arabica) transformation parameters using uidA and hpt genes: effect of osmotic pre-treatment, helium pressure and target distance
title_full	Optimization of coffee (Coffea arabica) transformation parameters using uidA and hpt genes: effect of osmotic pre-treatment, helium pressure and target distance
title_fullStr	Optimization of coffee (Coffea arabica) transformation parameters using uidA and hpt genes: effect of osmotic pre-treatment, helium pressure and target distance
title_full_unstemmed	Optimization of coffee (Coffea arabica) transformation parameters using uidA and hpt genes: effect of osmotic pre-treatment, helium pressure and target distance
title_sort	optimization of coffee (coffea arabica) transformation parameters using uida and hpt genes: effect of osmotic pre-treatment, helium pressure and target distance
title_alt	Optimization of coffee (Coffea arabica) transformation parameters using uidA and hpt genes: effect of osmotic pre-treatment, helium pressure and target distance
publisher	Universidad de Costa Rica
publishDate	2009
url	https://revistas.ucr.ac.cr/index.php/rbt/article/view/21292
work_keys_str_mv	AT gaticaandresm optimizationofcoffeecoffeaarabicatransformationparametersusinguidaandhptgeneseffectofosmoticpretreatmentheliumpressureandtargetdistance AT arrietagriselda optimizationofcoffeecoffeaarabicatransformationparametersusinguidaandhptgeneseffectofosmoticpretreatmentheliumpressureandtargetdistance AT espinozaanam optimizationofcoffeecoffeaarabicatransformationparametersusinguidaandhptgeneseffectofosmoticpretreatmentheliumpressureandtargetdistance
_version_	1810114832507076608

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

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