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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| Format: | Article |
| Language: | English English |
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Elsevier
2006
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| Online Access: | http://psasir.upm.edu.my/id/eprint/18366/ http://psasir.upm.edu.my/id/eprint/18366/2/Optimization%20of%20Agrobacterium.pdf |
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| author | Yong, Wilson Thau Lim Abdullah, Janna Ong Mahmood, Maziah |
| author_facet | Yong, Wilson Thau Lim Abdullah, Janna Ong Mahmood, Maziah |
| author_sort | Yong, Wilson Thau Lim |
| building | UPM Institutional Repository |
| collection | Online Access |
| 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 |
| first_indexed | 2025-11-15T08:15:53Z |
| format | Article |
| id | upm-18366 |
| institution | Universiti Putra Malaysia |
| institution_category | Local University |
| language | English English |
| last_indexed | 2025-11-15T08:15:53Z |
| publishDate | 2006 |
| publisher | Elsevier |
| recordtype | eprints |
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| spelling | upm-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 Melastomataceae - Biotechnology - Malaysia Cell transformation Green fluorescent protein doi:10.1016/j.scienta.2006.03.005 English |
| spellingShingle | Melastomataceae - Biotechnology - Malaysia Cell transformation Green fluorescent protein 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 |
| title | 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_short | 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 |
| topic | Melastomataceae - Biotechnology - Malaysia Cell transformation Green fluorescent protein |
| url | http://psasir.upm.edu.my/id/eprint/18366/ http://psasir.upm.edu.my/id/eprint/18366/ http://psasir.upm.edu.my/id/eprint/18366/ http://psasir.upm.edu.my/id/eprint/18366/2/Optimization%20of%20Agrobacterium.pdf |