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All Journal Prosiding Seminar Biologi AGRIVITA, Journal of Agricultural Science Indonesian Journal of Biotechnology Biosaintifika: Journal of Biology & Biology Education
Sukarti Moeljopawiro
Faculty of Biology, Gadjah Mada University
Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Immunology & microbiology Materials Science & Nanotechnology

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Journal : AGRIVITA,%20Journal%20of%20Agricultural%20Science

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Micropropagation of Dendrobium phalaenopsis Orchid Through Overexpression of Embryo Gene AtRKD4 Setiari, Nintya; Purwantoro, Aziz; Moeljopawiro, Sukarti; Semiarti, Endang
AGRIVITA, Journal of Agricultural Science Vol 40, No 2 (2018): JUNE
Publisher : Faculty of Agriculture University of Brawijaya in collaboration with PERAGI

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.17503/agrivita.v40i2.1690

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.
Co-Authors . HERNAYANTI Ahmad Hamim Sadewa Ari Indrianto Aziz Purwantoro Bambang Hariono Bambang Setiaji Endang Semiarti Erma Sulistyaningsih Galih G, Galih Jarot Subandono Kumala Dewi LANGKAH SEMBIRING Nintya Setiari Noor Aini Habibah Sarkono Sarkono Subagus Wahyuono Tri R Nuringtyas, Tri R Wayan T. Artama Yoga Pratama, Yoga