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Micropropagation of Dendrobium phalaenopsis Orchid Through Overexpression of Embryo Gene AtRKD4

AGRIVITA, Journal of Agricultural Science · Vol. 40 No. 2 (2018) · hlm. 284–294

DOI
10.17503/agrivita.v40i2.1690
ISSN
2477-8516, 0126-0537
Terbit
2018-04-27
Penerbit
Faculty of Agriculture University of Brawijaya in collaboration with PERAGI
Bahasa
eng
Lisensi
creativecommons.org/licenses/by-nc/4.0

Abstrak

To increase the efficiency of crop production from Dendrobium phalaenopsis orchids, mass propagation has been performed by inducing somatic embryogenesis through Agrobacterium-mediated transformation of the Arabidopsis embryo gene AtRKD4 into orchid protocorm (developing orchid embryo). The three-week-old protocorms of D. phalaenopsis were genetically transformed with T-DNA carrying 35S:: GAL4:: AtRKD4:: GR through A. tumefaciens strain EHA 105. The cultures were maintained in VW medium with 10 mg L-1 Hygromycin. Due to the existence of glucocorticoid response element (GR) in the T-DNA construct, the transformed protocorms were transferred into VW medium with the addition of 15 μM Dexamethasone in 6 weeks after transformation to activate the transgene. A total of 12% protocorms has been confirmed for Hyg + by using PCR. The expression of embryo gene AtRKD4 was confirmed by cDNA analysis using AtRKD4 specific primers and Actin primers as a positive control experiment. The expression level of AtRKD4 in 2.5-month-old D. phalaenopsis transformant shoots was 7 times higher than non-transformant plants, and increased to 86 times higher in 8-months, that much higher than that of non-transformant. These results provide an improved method for genetic transformation of D. Phalaenopsis and will (eventually) increase production efficiency in the future.

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Sumber metadata: OAI-PMH (metadata jurnal) https://agrivita.ub.ac.id/index.php/agrivita/oai, pertama di-harvest 8 Nov 2026, terakhir terlihat 8 Nov 2026.
Metadata asli (oai_dc) 
<oai_dc:dc xmlns="http://www.openarchives.org/OAI/2.0/" xmlns:dc="http://purl.org/dc/elements/1.1/" xmlns:oai_dc="http://www.openarchives.org/OAI/2.0/oai_dc/" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" xsi:schemaLocation="http://www.openarchives.org/OAI/2.0/oai_dc/  http://www.openarchives.org/OAI/2.0/oai_dc.xsd">
	<dc:title xml:lang="en-US">Micropropagation of Dendrobium phalaenopsis Orchid Through Overexpression of Embryo Gene AtRKD4</dc:title>
	<dc:creator>Setiari, Nintya</dc:creator>
	<dc:creator>Purwantoro, Aziz</dc:creator>
	<dc:creator>Moeljopawiro, Sukarti</dc:creator>
	<dc:creator>Semiarti, Endang</dc:creator>
	<dc:subject xml:lang="en-US"></dc:subject>
	<dc:subject xml:lang="en-US">AtRKD4; Dendrobium phalaenopsis; Glucocorticoid; Micropropagation; Somatic Embryo</dc:subject>
	<dc:description xml:lang="en-US">To increase the efficiency of crop production from Dendrobium phalaenopsis orchids, mass propagation has been performed by inducing somatic embryogenesis through Agrobacterium-mediated transformation of the Arabidopsis embryo gene AtRKD4 into orchid protocorm (developing orchid embryo). The three-week-old protocorms of D. phalaenopsis were genetically transformed with T-DNA carrying 35S :: GAL4 :: AtRKD4 :: GR through A. tumefaciens strain EHA 105. The cultures were maintained in VW medium with 10 mg L-1 Hygromycin. Due to the existence of glucocorticoid response element (GR) in the T-DNA construct, the transformed protocorms were transferred into VW medium with the addition of 15 μM Dexamethasone in 6 weeks after transformation to activate the transgene. A total of 12% protocorms has been confirmed for Hyg + by using PCR. The expression of embryo gene AtRKD4 was confirmed by cDNA analysis using AtRKD4 specific primers and Actin primers as a positive control experiment. The expression level of AtRKD4 in 2.5-month-old D. phalaenopsis transformant shoots was 7 times higher than non-transformant plants, and increased to 86 times higher in 8-months, that much higher than that of non-transformant. These results provide an improved method for genetic transformation of D. Phalaenopsis and will (eventually) increase production efficiency in the future.</dc:description>
	<dc:publisher xml:lang="en-US">Faculty of Agriculture University of Brawijaya in collaboration with PERAGI</dc:publisher>
	<dc:contributor xml:lang="en-US">Dr. Luke Daniels,</dc:contributor>
	<dc:contributor xml:lang="en-US">PUPT Grant, Ministry of Research Technology and Higher Education Republic Indonesia</dc:contributor>
	<dc:contributor xml:lang="en-US">Doctoral Grant, Universitas Diponegoro</dc:contributor>
	<dc:date>2018-04-27</dc:date>
	<dc:type>info:eu-repo/semantics/article</dc:type>
	<dc:type>info:eu-repo/semantics/publishedVersion</dc:type>
	<dc:type xml:lang="en-US">Peer-reviewed Article</dc:type>
	<dc:format>application/pdf</dc:format>
	<dc:identifier>https://agrivita.ub.ac.id/index.php/agrivita/article/view/1690</dc:identifier>
	<dc:identifier>10.17503/agrivita.v40i2.1690</dc:identifier>
	<dc:source xml:lang="en-US">AGRIVITA Journal of Agricultural Science; Vol 40, No 2 (2018): JUNE; 284-294</dc:source>
	<dc:source>2477-8516</dc:source>
	<dc:source>0126-0537</dc:source>
	<dc:source>10.17503/agrivita.v40i2</dc:source>
	<dc:language>eng</dc:language>
	<dc:relation>https://agrivita.ub.ac.id/index.php/agrivita/article/view/1690/933</dc:relation>
	<dc:relation>https://agrivita.ub.ac.id/index.php/agrivita/article/downloadSuppFile/1690/860</dc:relation>
	<dc:relation>https://agrivita.ub.ac.id/index.php/agrivita/article/downloadSuppFile/1690/959</dc:relation>
	<dc:rights xml:lang="en-US">Copyright (c) 2018 UNIVERSITAS BRAWIJAYA</dc:rights>
	<dc:rights xml:lang="en-US">http://creativecommons.org/licenses/by-nc/4.0</dc:rights>
</oai_dc:dc>