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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) · pp. 284–294

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

Abstract

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.

Keywords

Collaboration network of the authors

40 authors · 100 co-authorship links · showing the 40 most connected

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  • Universitas Gadjah Mada
  • Affiliation unknown
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Co-authorOrganisationJoint articles
Ari IndriantoAffiliation unknown7
Muhammad Dylan LawrieAffiliation unknown7
Exsyupransia MursyantiAffiliation unknown6
Agus Budi SetiawanAffiliation unknown5
Windi MoseAffiliation unknown4
Yulita NurchayatiAffiliation unknown3
Febri Yuda KurniawanAffiliation unknown3
Jose Gutierrez-MarcosAffiliation unknown3
Jaka WidadaUniversitas Gadjah Mada3
Aries Bagus SasongkoUniversitas Gadjah Mada3
Endang SulistyaningsihAffiliation unknown2
Dyah Weny RespatieAffiliation unknown2
Chee How TeoAffiliation unknown2
Aviesta LinggabuwanaAffiliation unknown2
Saifa Usni PutriAffiliation unknown2
Nuzlan RasjidAffiliation unknown2
Sismindari SismindariAffiliation unknown2
Fitriana PuspitasariAffiliation unknown2
Naufal Ghozi Aditya PerdanaAffiliation unknown2
Dika SundariAffiliation unknown2
Triono Bagus SaputroAffiliation unknown2
Erma PrihastantiAffiliation unknown1
Maria UlvaAffiliation unknown1
Khoirin NidaAffiliation unknown1
Masrukhatul LuaeliyahAffiliation unknown1
Munifatul IzzatiAffiliation unknown1
Asadi AsadiAffiliation unknown1
Sahiral YakubAffiliation unknown1
Ari WibowoAffiliation unknown1
Maria Marina HerawatiAffiliation unknown1
Endang PudjihartatiAffiliation unknown1
Suwijiyo PramonoAffiliation unknown1
Tri Joko RaharjoAffiliation unknown1
Stalis Norma EthicaAffiliation unknown1
Oedjijono OedjijonoAffiliation unknown1
Irna AuliauzzakiaAffiliation unknown1

Affiliations come from OpenAlex; journal OAI-PMH metadata carries none. Authors without an OpenAlex ID are matched by name, which can merge or split people.

Metadata source: OAI-PMH (metadata jurnal) https://agrivita.ub.ac.id/index.php/agrivita/oai, first harvested 8 Nov 2026, last seen 8 Nov 2026.
Original metadata (oai_dc) 
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	<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>