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

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 Murashig...

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Main Authors:	Chew, Yee-Chern, Wan Muhamad Asrul Nizam Wan Abdullah, Kok, Andrew De-Xian, Janna Ong-Abdullah, Maziah Mahmood, Lai, Kok-Song
Format:	Article
Language:	English
Published:	Penerbit Universiti Kebangsaan Malaysia 2019
Online Access:	http://journalarticle.ukm.my/14340/ http://journalarticle.ukm.my/14340/ http://journalarticle.ukm.my/14340/1/07%20Yee-Chern%20Chew.pdf

id	ukm-14340
recordtype	eprints
spelling	ukm-143402020-03-03T07:58:00Z http://journalarticle.ukm.my/14340/ Development of an efficient particle bombardment transformation system for the endemic orchid, Phalaenopsis bellina Chew, Yee-Chern Wan Muhamad Asrul Nizam Wan Abdullah, Kok, Andrew De-Xian Janna Ong-Abdullah, Maziah Mahmood, Lai, Kok-Song 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, 27 mmHg 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. Penerbit Universiti Kebangsaan Malaysia 2019-09 Article PeerReviewed application/pdf en http://journalarticle.ukm.my/14340/1/07%20Yee-Chern%20Chew.pdf Chew, Yee-Chern and Wan Muhamad Asrul Nizam Wan Abdullah, and Kok, Andrew De-Xian and Janna Ong-Abdullah, and Maziah Mahmood, 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 http://www.ukm.my/jsm/malay_journals/jilid48bil9_2019/KandunganJilid48Bil9_2019.html
repository_type	Digital Repository
institution_category	Local University
institution	Universiti Kebangasaan Malaysia
building	UKM Institutional Repository
collection	Online Access
language	English
description	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, 27 mmHg 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.
format	Article
author	Chew, Yee-Chern Wan Muhamad Asrul Nizam Wan Abdullah, Kok, Andrew De-Xian Janna Ong-Abdullah, Maziah Mahmood, Lai, Kok-Song
spellingShingle	Chew, Yee-Chern Wan Muhamad Asrul Nizam Wan Abdullah, Kok, Andrew De-Xian Janna Ong-Abdullah, Maziah Mahmood, Lai, Kok-Song Development of an efficient particle bombardment transformation system for the endemic orchid, Phalaenopsis bellina
author_facet	Chew, Yee-Chern Wan Muhamad Asrul Nizam Wan Abdullah, Kok, Andrew De-Xian Janna Ong-Abdullah, Maziah Mahmood, Lai, Kok-Song
author_sort	Chew, Yee-Chern
title	Development of an efficient particle bombardment transformation system for the endemic orchid, Phalaenopsis bellina
title_short	Development of an efficient particle bombardment transformation system for the endemic orchid, Phalaenopsis bellina
title_full	Development of an efficient particle bombardment transformation system for the endemic orchid, Phalaenopsis bellina
title_fullStr	Development of an efficient particle bombardment transformation system for the endemic orchid, Phalaenopsis bellina
title_full_unstemmed	Development of an efficient particle bombardment transformation system for the endemic orchid, Phalaenopsis bellina
title_sort	development of an efficient particle bombardment transformation system for the endemic orchid, phalaenopsis bellina
publisher	Penerbit Universiti Kebangsaan Malaysia
publishDate	2019
url	http://journalarticle.ukm.my/14340/ http://journalarticle.ukm.my/14340/ http://journalarticle.ukm.my/14340/1/07%20Yee-Chern%20Chew.pdf
first_indexed	2023-09-18T20:06:52Z
last_indexed	2023-09-18T20:06:52Z
_version_	1777407217328193536

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

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