Optimization in Agrobacterium-mediated transformation of Anthurium andraeanum using GFP as a reporter

Although Agrobacterium-mediated transformation protocols for many economically important plant species have been well established, protocol for a number of flowering plants including Anthurium andraeanum remains challenging. In this study, we report success in generating transgenic Anthurium andraea...

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Autores principales:	Zhao,Qing, Jing,Ji, Wang,Gang, Wang,Jie Hua, Feng,Yuan Yuan, Xing,Han Wen, Guan,Chun Feng
Lenguaje:	English
Publicado:	Pontificia Universidad Católica de Valparaíso 2010
Materias:	cocultivation gene transfer green fluorescent protein regeneration
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582010000500009
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spelling	oai:scielo:S0717-345820100005000092011-05-24Optimization in Agrobacterium-mediated transformation of Anthurium andraeanum using GFP as a reporterZhao,QingJing,JiWang,GangWang,Jie HuaFeng,Yuan YuanXing,Han WenGuan,Chun Feng cocultivation gene transfer green fluorescent protein regeneration Although Agrobacterium-mediated transformation protocols for many economically important plant species have been well established, protocol for a number of flowering plants including Anthurium andraeanum remains challenging. In this study, we report success in generating transgenic Anthurium andraeanum cv Arizona using Agrobacterium GV3101 strain harboring a binary vector carrying gfp as a reporter gene. The possibility of facilitating the screening process for transgenic plants expressing functional proteins using gfp marker was explored. In order to realize high transformation efficiency, different explant sources including undifferentiated callus pieces and petioles were compared for their regeneration efficiency and susceptibility to Agrobacterium-mediated transformation. We also optimized the concentration of AS added to co-cultivation media. Genomic PCR revealed that 11 of the 22 resistant plantlets regenerated on selective medium were successfully transformed. Green fluorescence was observed using a fluorescence microscope in 7 of the 11 PCR-positive plants, indicating GFP was expressed stably in the transformed Anthurium andraeanum. The highest transformation efficiency obtained in this study was 1.71% (percentage of explants with transgenic shoots in total explants) when callus explants were used as starting material and 125 μmol l-1 AS was added during the co-cultivation process.info:eu-repo/semantics/openAccessPontificia Universidad Católica de ValparaísoElectronic Journal of Biotechnology v.13 n.5 20102010-09-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582010000500009en
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	cocultivation gene transfer green fluorescent protein regeneration
spellingShingle	cocultivation gene transfer green fluorescent protein regeneration Zhao,Qing Jing,Ji Wang,Gang Wang,Jie Hua Feng,Yuan Yuan Xing,Han Wen Guan,Chun Feng Optimization in Agrobacterium-mediated transformation of Anthurium andraeanum using GFP as a reporter
description	Although Agrobacterium-mediated transformation protocols for many economically important plant species have been well established, protocol for a number of flowering plants including Anthurium andraeanum remains challenging. In this study, we report success in generating transgenic Anthurium andraeanum cv Arizona using Agrobacterium GV3101 strain harboring a binary vector carrying gfp as a reporter gene. The possibility of facilitating the screening process for transgenic plants expressing functional proteins using gfp marker was explored. In order to realize high transformation efficiency, different explant sources including undifferentiated callus pieces and petioles were compared for their regeneration efficiency and susceptibility to Agrobacterium-mediated transformation. We also optimized the concentration of AS added to co-cultivation media. Genomic PCR revealed that 11 of the 22 resistant plantlets regenerated on selective medium were successfully transformed. Green fluorescence was observed using a fluorescence microscope in 7 of the 11 PCR-positive plants, indicating GFP was expressed stably in the transformed Anthurium andraeanum. The highest transformation efficiency obtained in this study was 1.71% (percentage of explants with transgenic shoots in total explants) when callus explants were used as starting material and 125 μmol l-1 AS was added during the co-cultivation process.
author	Zhao,Qing Jing,Ji Wang,Gang Wang,Jie Hua Feng,Yuan Yuan Xing,Han Wen Guan,Chun Feng
author_facet	Zhao,Qing Jing,Ji Wang,Gang Wang,Jie Hua Feng,Yuan Yuan Xing,Han Wen Guan,Chun Feng
author_sort	Zhao,Qing
title	Optimization in Agrobacterium-mediated transformation of Anthurium andraeanum using GFP as a reporter
title_short	Optimization in Agrobacterium-mediated transformation of Anthurium andraeanum using GFP as a reporter
title_full	Optimization in Agrobacterium-mediated transformation of Anthurium andraeanum using GFP as a reporter
title_fullStr	Optimization in Agrobacterium-mediated transformation of Anthurium andraeanum using GFP as a reporter
title_full_unstemmed	Optimization in Agrobacterium-mediated transformation of Anthurium andraeanum using GFP as a reporter
title_sort	optimization in agrobacterium-mediated transformation of anthurium andraeanum using gfp as a reporter
publisher	Pontificia Universidad Católica de Valparaíso
publishDate	2010
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582010000500009
work_keys_str_mv	AT zhaoqing optimizationinagrobacteriummediatedtransformationofanthuriumandraeanumusinggfpasareporter AT jingji optimizationinagrobacteriummediatedtransformationofanthuriumandraeanumusinggfpasareporter AT wanggang optimizationinagrobacteriummediatedtransformationofanthuriumandraeanumusinggfpasareporter AT wangjiehua optimizationinagrobacteriummediatedtransformationofanthuriumandraeanumusinggfpasareporter AT fengyuanyuan optimizationinagrobacteriummediatedtransformationofanthuriumandraeanumusinggfpasareporter AT xinghanwen optimizationinagrobacteriummediatedtransformationofanthuriumandraeanumusinggfpasareporter AT guanchunfeng optimizationinagrobacteriummediatedtransformationofanthuriumandraeanumusinggfpasareporter
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Optimization in Agrobacterium-mediated transformation of Anthurium andraeanum using GFP as a reporter

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