Development of pollen mediated activation tagging system for Phalaenopsis and Doritaenopsis

In the present study, a novel plant transformation system for Doritaenopsis and Phalaenopsis has been developed. The pollen-mediated activation tagging system was established by artificial pollination. The pollens, co-cultured with Agrobacterium tumefaciens strain EHA105 harbouring an activation tag...

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Autores principales:	Tsay,Hsin-Sheng, Ho,Hui-Min, Gupta,Sushim K, Wang,Chang-Sheng, Chen,Pei-Tsu, Chen,Emily Chin-Fun
Lenguaje:	English
Publicado:	Pontificia Universidad Católica de Valparaíso 2012
Materias:	activation tagging Agrobacterium tumefaciens Doritaenopsis cultivars Phalaenopsis amabilis transformation
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582012000400009
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spelling	oai:scielo:S0717-345820120004000092012-08-22Development of pollen mediated activation tagging system for Phalaenopsis and DoritaenopsisTsay,Hsin-ShengHo,Hui-MinGupta,Sushim KWang,Chang-ShengChen,Pei-TsuChen,Emily Chin-Fun activation tagging Agrobacterium tumefaciens Doritaenopsis cultivars Phalaenopsis amabilis transformation In the present study, a novel plant transformation system for Doritaenopsis and Phalaenopsis has been developed. The pollen-mediated activation tagging system was established by artificial pollination. The pollens, co-cultured with Agrobacterium tumefaciens strain EHA105 harbouring an activation tagging vector (pTAG-8), were used for pollination. In order to optimize the transformation efficiency, several factors (concentration of A. tumefaciens, concentration of acetosyringone during co-cultivation and the duration of co-cultivation) known to influence Agrobacterium-mediated DNA transfer were examined. A concentration of 0.5-1 x 10(8) CFU/ml for A. tumefaciens, 0.1 mM acetosyringone, and 6 hrs of co-culture period were found to be the optimal condition for high transformation efficiency. Integration of T-DNA into the genome of putative transgenic plants was confirmed by PCR and DNA blot analyses. Single copy of the transgene was observed in all transgenic plants analyzed. Most of the transgenic plants had a morphologically normal phenotype and the overall capsule formation efficiency was similar to control plant. Our results showed a new approach of genetic transformation in orchids and this method can be employed for genetic improvement of the orchids.info:eu-repo/semantics/openAccessPontificia Universidad Católica de ValparaísoElectronic Journal of Biotechnology v.15 n.4 20122012-07-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582012000400009en
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	activation tagging Agrobacterium tumefaciens Doritaenopsis cultivars Phalaenopsis amabilis transformation
spellingShingle	activation tagging Agrobacterium tumefaciens Doritaenopsis cultivars Phalaenopsis amabilis transformation Tsay,Hsin-Sheng Ho,Hui-Min Gupta,Sushim K Wang,Chang-Sheng Chen,Pei-Tsu Chen,Emily Chin-Fun Development of pollen mediated activation tagging system for Phalaenopsis and Doritaenopsis
description	In the present study, a novel plant transformation system for Doritaenopsis and Phalaenopsis has been developed. The pollen-mediated activation tagging system was established by artificial pollination. The pollens, co-cultured with Agrobacterium tumefaciens strain EHA105 harbouring an activation tagging vector (pTAG-8), were used for pollination. In order to optimize the transformation efficiency, several factors (concentration of A. tumefaciens, concentration of acetosyringone during co-cultivation and the duration of co-cultivation) known to influence Agrobacterium-mediated DNA transfer were examined. A concentration of 0.5-1 x 10(8) CFU/ml for A. tumefaciens, 0.1 mM acetosyringone, and 6 hrs of co-culture period were found to be the optimal condition for high transformation efficiency. Integration of T-DNA into the genome of putative transgenic plants was confirmed by PCR and DNA blot analyses. Single copy of the transgene was observed in all transgenic plants analyzed. Most of the transgenic plants had a morphologically normal phenotype and the overall capsule formation efficiency was similar to control plant. Our results showed a new approach of genetic transformation in orchids and this method can be employed for genetic improvement of the orchids.
author	Tsay,Hsin-Sheng Ho,Hui-Min Gupta,Sushim K Wang,Chang-Sheng Chen,Pei-Tsu Chen,Emily Chin-Fun
author_facet	Tsay,Hsin-Sheng Ho,Hui-Min Gupta,Sushim K Wang,Chang-Sheng Chen,Pei-Tsu Chen,Emily Chin-Fun
author_sort	Tsay,Hsin-Sheng
title	Development of pollen mediated activation tagging system for Phalaenopsis and Doritaenopsis
title_short	Development of pollen mediated activation tagging system for Phalaenopsis and Doritaenopsis
title_full	Development of pollen mediated activation tagging system for Phalaenopsis and Doritaenopsis
title_fullStr	Development of pollen mediated activation tagging system for Phalaenopsis and Doritaenopsis
title_full_unstemmed	Development of pollen mediated activation tagging system for Phalaenopsis and Doritaenopsis
title_sort	development of pollen mediated activation tagging system for phalaenopsis and doritaenopsis
publisher	Pontificia Universidad Católica de Valparaíso
publishDate	2012
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582012000400009
work_keys_str_mv	AT tsayhsinsheng developmentofpollenmediatedactivationtaggingsystemforphalaenopsisanddoritaenopsis AT hohuimin developmentofpollenmediatedactivationtaggingsystemforphalaenopsisanddoritaenopsis AT guptasushimk developmentofpollenmediatedactivationtaggingsystemforphalaenopsisanddoritaenopsis AT wangchangsheng developmentofpollenmediatedactivationtaggingsystemforphalaenopsisanddoritaenopsis AT chenpeitsu developmentofpollenmediatedactivationtaggingsystemforphalaenopsisanddoritaenopsis AT chenemilychinfun developmentofpollenmediatedactivationtaggingsystemforphalaenopsisanddoritaenopsis
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Development of pollen mediated activation tagging system for Phalaenopsis and Doritaenopsis

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