Micropropagation of Dendrobium phalaenopsis Orchid Through Overexpression of Embryo Gene AtRKD4

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-we...

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Main Authors: Setiari, Nintya (Author), Purwantoro, Aziz (Author), Moeljopawiro, Sukarti (Author), Semiarti, Endang (Author)
Other Authors: Dr. Luke Daniels (Contributor), PUPT Grant, Ministry of Research Technology and Higher Education Republic Indonesia (Contributor), Doctoral Grant, Universitas Diponegoro (Contributor)
Format: EJournal Article
Published: Faculty of Agriculture University of Brawijaya in collaboration with PERAGI, 2018-04-27.
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042 |a dc 
100 1 0 |a Setiari, Nintya  |e author 
100 1 0 |a Dr. Luke Daniels,  |e contributor 
100 1 0 |a PUPT Grant, Ministry of Research Technology and Higher Education Republic Indonesia  |e contributor 
100 1 0 |a Doctoral Grant, Universitas Diponegoro  |e contributor 
700 1 0 |a Purwantoro, Aziz  |e author 
700 1 0 |a Moeljopawiro, Sukarti  |e author 
700 1 0 |a Semiarti, Endang  |e author 
245 0 0 |a Micropropagation of Dendrobium phalaenopsis Orchid Through Overexpression of Embryo Gene AtRKD4 
260 |b Faculty of Agriculture University of Brawijaya in collaboration with PERAGI,   |c 2018-04-27. 
500 |a https://agrivita.ub.ac.id/index.php/agrivita/article/view/1690 
520 |a 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. 
540 |a Copyright (c) 2018 UNIVERSITAS BRAWIJAYA 
540 |a http://creativecommons.org/licenses/by-nc/4.0 
546 |a eng 
690
690 |a AtRKD4; Dendrobium phalaenopsis; Glucocorticoid; Micropropagation; Somatic Embryo 
655 7 |a info:eu-repo/semantics/article  |2 local 
655 7 |a info:eu-repo/semantics/publishedVersion  |2 local 
655 7 |a Peer-reviewed Article  |2 local 
786 0 |n AGRIVITA, Journal of Agricultural Science; Vol 40, No 2 (2018): JUNE; 284-294 
786 0 |n 2477-8516 
786 0 |n 0126-0537 
786 0 |n 10.17503/agrivita.v40i2 
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