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Optimization of Agrobacterium-mediated transformation parameters for Melastomataceae spp. using green fluorescent protein (GFP) as a reporter

Agrobacterium-mediated transformation for both Melastoma malabathricum and Tibouchina semidecandra were optimized using green fluorescent protein (GFP) as a reporter. The binary vector pCAMBIA1304 harboring the modified green fluorescent protein (mgfp) gene driven by the CaMV 35S promoter was used....

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Main Authors: Yong, Wilson Thau Lim, Abdullah, Janna Ong, Mahmood, Maziah
Format: Article
Language:English
English
Published: Elsevier 2006
Online Access:http://psasir.upm.edu.my/id/eprint/18366/2/Optimization%20of%20Agrobacterium.pdf
http://psasir.upm.edu.my/id/eprint/18366/
http://dx.doi.org/10.1016/j.scienta.2006.03.005
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spelling my.upm.eprints.183662015-11-11T03:38:27Z http://psasir.upm.edu.my/id/eprint/18366/ Optimization of Agrobacterium-mediated transformation parameters for Melastomataceae spp. using green fluorescent protein (GFP) as a reporter Yong, Wilson Thau Lim Abdullah, Janna Ong Mahmood, Maziah Agrobacterium-mediated transformation for both Melastoma malabathricum and Tibouchina semidecandra were optimized using green fluorescent protein (GFP) as a reporter. The binary vector pCAMBIA1304 harboring the modified green fluorescent protein (mgfp) gene driven by the CaMV 35S promoter was used. Parameters optimized were bacterial strain, bacterial concentration, pre-culture period, co-cultivation period, immersion time, acetosyringone concentration and wounding type. Results obtained were based on the percentage of GFP expression which was observed 3 days post-transformation. Agrobacterium tumefaciens strain LBA4404 and EHA105 at concentration 1 × 107 cfu ml-1 (OD600nm 0.8) showed the highest virulence on M. malabathricum and T. semidecandra, respectively. Four days of pre-culture and 2 days of co-cultivation were optimum for M. malabathricum transformation, while 3 days of pre-culture and co-cultivation for T. semidecandra. Result also showed that 60 min of immersion and addition of 200 μM acetosyringone gave the highest percentage of positive transformants for both M. malabathricum and T. semidecandra. Mild wounding also significantly increased the efficiency of M. malabathricum transformation. © 2006 Elsevier B.V. All rights reserved Elsevier 2006-06 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/18366/2/Optimization%20of%20Agrobacterium.pdf Yong, Wilson Thau Lim and Abdullah, Janna Ong and Mahmood, Maziah (2006) Optimization of Agrobacterium-mediated transformation parameters for Melastomataceae spp. using green fluorescent protein (GFP) as a reporter. Scientia Horticulturae, 109 (1). pp. 78-85. ISSN 0304--4238 http://dx.doi.org/10.1016/j.scienta.2006.03.005 doi:10.1016/j.scienta.2006.03.005 English
institution Universiti Putra Malaysia
building UPM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Putra Malaysia
content_source UPM Institutional Repository
url_provider http://psasir.upm.edu.my/
language English
English
description Agrobacterium-mediated transformation for both Melastoma malabathricum and Tibouchina semidecandra were optimized using green fluorescent protein (GFP) as a reporter. The binary vector pCAMBIA1304 harboring the modified green fluorescent protein (mgfp) gene driven by the CaMV 35S promoter was used. Parameters optimized were bacterial strain, bacterial concentration, pre-culture period, co-cultivation period, immersion time, acetosyringone concentration and wounding type. Results obtained were based on the percentage of GFP expression which was observed 3 days post-transformation. Agrobacterium tumefaciens strain LBA4404 and EHA105 at concentration 1 × 107 cfu ml-1 (OD600nm 0.8) showed the highest virulence on M. malabathricum and T. semidecandra, respectively. Four days of pre-culture and 2 days of co-cultivation were optimum for M. malabathricum transformation, while 3 days of pre-culture and co-cultivation for T. semidecandra. Result also showed that 60 min of immersion and addition of 200 μM acetosyringone gave the highest percentage of positive transformants for both M. malabathricum and T. semidecandra. Mild wounding also significantly increased the efficiency of M. malabathricum transformation. © 2006 Elsevier B.V. All rights reserved
format Article
author Yong, Wilson Thau Lim
Abdullah, Janna Ong
Mahmood, Maziah
spellingShingle Yong, Wilson Thau Lim
Abdullah, Janna Ong
Mahmood, Maziah
Optimization of Agrobacterium-mediated transformation parameters for Melastomataceae spp. using green fluorescent protein (GFP) as a reporter
author_facet Yong, Wilson Thau Lim
Abdullah, Janna Ong
Mahmood, Maziah
author_sort Yong, Wilson Thau Lim
title Optimization of Agrobacterium-mediated transformation parameters for Melastomataceae spp. using green fluorescent protein (GFP) as a reporter
title_short Optimization of Agrobacterium-mediated transformation parameters for Melastomataceae spp. using green fluorescent protein (GFP) as a reporter
title_full Optimization of Agrobacterium-mediated transformation parameters for Melastomataceae spp. using green fluorescent protein (GFP) as a reporter
title_fullStr Optimization of Agrobacterium-mediated transformation parameters for Melastomataceae spp. using green fluorescent protein (GFP) as a reporter
title_full_unstemmed Optimization of Agrobacterium-mediated transformation parameters for Melastomataceae spp. using green fluorescent protein (GFP) as a reporter
title_sort optimization of agrobacterium-mediated transformation parameters for melastomataceae spp. using green fluorescent protein (gfp) as a reporter
publisher Elsevier
publishDate 2006
url http://psasir.upm.edu.my/id/eprint/18366/2/Optimization%20of%20Agrobacterium.pdf
http://psasir.upm.edu.my/id/eprint/18366/
http://dx.doi.org/10.1016/j.scienta.2006.03.005
_version_ 1643826783170068480
score 13.211869

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