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

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

Full text not available from this repository.

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

Abstract

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.

Item Type:Article
Keyword:Agrobacterium, Melastomataceae, Green fluorescent protein
Faculty or Institute:Faculty of Biotechnology and Biomolecular Sciences
Publisher:Elsevier
DOI Number:10.1016/j.scienta.2006.03.005
ID Code:713
Deposited By:INVALID USER
Deposited On:25 Nov 2008 12:57
Last Modified:30 Oct 2011 05:06

Repository Staff Only: item control page


Universiti Putra Malaysia Institutional Repository

Universiti Putra Malaysia Institutional Repository is an on-line digital archive that serves as a central collection and storage of scientific information and research at the Universiti Putra Malaysia.

Currently, the collections deposited in the IR consists of Master and PhD theses, Master and PhD Project Report, Journal Articles, Journal Bulletins, Conference Papers, UPM News, Newspaper Cuttings, Patents and Inaugural Lectures.

As the policy of the university does not permit users to view thesis in full text, access is only given to the first 24 pages only.