Optimization of Agrobacterium-Mediated Transformation Parameters for Melastomatacea Spp. Using Green Fluorescent Protein(GFP) as A Reporter

Agrobacterium-mediated transformation for both Melastoma malabathricum and Tibouchinu semidecanda 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 CuMV 35S promoter was used. P...

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Main Authors: Thau, Wilson Lym Yong, Abdullah, Janna Ong, Mahmood, Maziah
Format: Article
Language:English
English
Published: Elsevier 2006
Online Access:http://psasir.upm.edu.my/id/eprint/713/
http://psasir.upm.edu.my/id/eprint/713/1/Optimization%20of%20Agrobacterium.pdf
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author Thau, Wilson Lym Yong
Abdullah, Janna Ong
Mahmood, Maziah
author_facet Thau, Wilson Lym Yong
Abdullah, Janna Ong
Mahmood, Maziah
author_sort Thau, Wilson Lym Yong
building UPM Institutional Repository
collection Online Access
description Agrobacterium-mediated transformation for both Melastoma malabathricum and Tibouchinu semidecanda 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 CuMV 35S promoter was used. Parameters optimized were bacterial strain, bacterial concentration, pre-culture period, co-cultivation period,immersion time, acetosyring concentration and wounding type. Results obtained obtained were based on the percentage of (GFP expression which was observed 3 days post-transformation. Agrobacterium tumefaciens starin LBA4404 and EHA105 at concentration 1 X 107 cfu ml -1 (OD 600mm 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 fot T, semidecandra, . result also showed that 60 min of immersion and addition of 200 p.M acetosyringone gave the highest percentage of positive transformants for both M. malabathricum and T.semidecandra. Mild wounding also significantly increased the efficiency 0f M.malabathricum transformation.
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spelling upm-7132016-01-21T01:58:37Z http://psasir.upm.edu.my/id/eprint/713/ Optimization of Agrobacterium-Mediated Transformation Parameters for Melastomatacea Spp. Using Green Fluorescent Protein(GFP) as A Reporter Thau, Wilson Lym Yong Abdullah, Janna Ong Mahmood, Maziah Agrobacterium-mediated transformation for both Melastoma malabathricum and Tibouchinu semidecanda 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 CuMV 35S promoter was used. Parameters optimized were bacterial strain, bacterial concentration, pre-culture period, co-cultivation period,immersion time, acetosyring concentration and wounding type. Results obtained obtained were based on the percentage of (GFP expression which was observed 3 days post-transformation. Agrobacterium tumefaciens starin LBA4404 and EHA105 at concentration 1 X 107 cfu ml -1 (OD 600mm 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 fot T, semidecandra, . result also showed that 60 min of immersion and addition of 200 p.M acetosyringone gave the highest percentage of positive transformants for both M. malabathricum and T.semidecandra. Mild wounding also significantly increased the efficiency 0f M.malabathricum transformation. Elsevier 2006 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/713/1/Optimization%20of%20Agrobacterium.pdf Thau, Wilson Lym Yong and Abdullah, Janna Ong and Mahmood, Maziah (2006) Optimization of Agrobacterium-Mediated Transformation Parameters for Melastomatacea 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 10.1016/j.scienta.2006.03.005 English
spellingShingle Thau, Wilson Lym Yong
Abdullah, Janna Ong
Mahmood, Maziah
Optimization of Agrobacterium-Mediated Transformation Parameters for Melastomatacea Spp. Using Green Fluorescent Protein(GFP) as A Reporter
title Optimization of Agrobacterium-Mediated Transformation Parameters for Melastomatacea Spp. Using Green Fluorescent Protein(GFP) as A Reporter
title_full Optimization of Agrobacterium-Mediated Transformation Parameters for Melastomatacea Spp. Using Green Fluorescent Protein(GFP) as A Reporter
title_fullStr Optimization of Agrobacterium-Mediated Transformation Parameters for Melastomatacea Spp. Using Green Fluorescent Protein(GFP) as A Reporter
title_full_unstemmed Optimization of Agrobacterium-Mediated Transformation Parameters for Melastomatacea Spp. Using Green Fluorescent Protein(GFP) as A Reporter
title_short Optimization of Agrobacterium-Mediated Transformation Parameters for Melastomatacea Spp. Using Green Fluorescent Protein(GFP) as A Reporter
title_sort optimization of agrobacterium-mediated transformation parameters for melastomatacea spp. using green fluorescent protein(gfp) as a reporter
url http://psasir.upm.edu.my/id/eprint/713/
http://psasir.upm.edu.my/id/eprint/713/
http://psasir.upm.edu.my/id/eprint/713/
http://psasir.upm.edu.my/id/eprint/713/1/Optimization%20of%20Agrobacterium.pdf