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Development of an efficient particle bombardment transformation system for the endemic orchid, Phalaenopsis bellina


Chew, Yee Chern and Wan Abdullah, Wan Muhamad Asrul Nizam and Kok, Andrew De-Xian and Abdullah, Janna Ong and Mahmood, Maziah and Lai, Kok Song (2019) Development of an efficient particle bombardment transformation system for the endemic orchid, Phalaenopsis bellina. Sains Malaysiana, 48 (9). pp. 1867-1877. ISSN 0126-6039


Phalaenopsis bellina is an important indigenous orchid with high commercial value. In this study, we established an efficient particle bombardment transformation system for P. bellina using the protocorm-like-bodies (PLBs) as target tissues. Leaf derived PLBs were proliferated on ½ strength Murashige and Skoog (MS) medium supplemented with 0.8 µM 2,4 dichlorophenoxyacetic acid (2, 4-D). Both physical and biological parameters affecting the transformation system were optimised using the green-fluorescent protein (GFP) and β-glucuronidase (GUS) as reporter systems. Optimal bombardment conditions with 6 cm target tissues distance, 1100 psi acceleration pressure, 1.0 μm gold particle size, 27mmHg chamber vacuum pressure, single bombardment time, spermidine as DNA precipitation agent, 72 h post bombardment incubation time, 2 μg plasmid DNA in 0.15:0.12 pmol ratio (pSMCHS:p35SGFP) were successfully determined. Surviving PLBs transformants were successfully recovered from the hygromycin selection medium and verified using genomic PCR analysis. The established system is not only useful for a simple and reliable transient gene analysis but as well as generating stable transformants for selective traits improvement in orchids.

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Additional Metadata

Item Type: Article
Divisions: Faculty of Biotechnology and Biomolecular Sciences
DOI Number: https://doi.org/10.17576/jsm-2019-4809-07
Publisher: Penerbit Universiti Kebangsaan Malaysia
Keywords: β-glucuronidase (GUS); Green fluorescent protein (GFP); Particle bombardment; Phalaenopsis bellina; Protocorm-like-bodies (PLBs)
Depositing User: Ms. Nuraida Ibrahim
Date Deposited: 09 Sep 2020 07:09
Last Modified: 09 Sep 2020 07:10
Altmetrics: http://www.altmetric.com/details.php?domain=psasir.upm.edu.my&doi=10.17576/jsm-2019-4809-07
URI: http://psasir.upm.edu.my/id/eprint/81147
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