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

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

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Official URL: http://dx.doi.org/10.1016/j.scienta.2006.03.005

Abstract

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

Item Type:Article
Keyword:Agrobacterium; Green fluorescent protein; Melastomataceae
Subject:Melastomataceae - Biotechnology - Malaysia
Subject:Cell transformation
Subject:Green fluorescent protein
Faculty or Institute:Faculty of Biotechnology and Biomolecular Sciences
Publisher:Elsevier
DOI Number:10.1016/j.scienta.2006.03.005
Altmetrics:http://www.altmetric.com/details.php?domain=psasir.upm.edu.my&doi=10.1016/j.scienta.2006.03.005
ID Code:18366
Deposited By: Azwana Abdul Rahman
Deposited On:30 Oct 2011 03:24
Last Modified:30 Oct 2011 03:24

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