Detection of AtRKD4 Protein During Induction of Somatic Embryogenesis in Dendrobium lineale Rolfe Transgenic Orchids Carrying 35S::GR::AtRKD4

https://doi.org/10.22146/jtbb.61783

Gde Cahyadi Wirajagat(1), Ni Luh Putu Kayika Febryanti(2), Fitriana Puspitasari(3), Dika Sundari(4), Jose Gutierrez-Marcos(5), Endang Semiarti(6*)

(1) Faculty of Biology, Universitas Gadjah Mada
(2) Faculty of Biology, Universitas Gadjah Mada
(3) Faculty of Biology, Universitas Gadjah Mada
(4) Faculty of Biology, Universitas Gadjah Mada
(5) School of Life Science, University of Warwick
(6) Faculty of Biology, Universitas Gadjah Mada
(*) Corresponding Author

Abstract


Dendrobium lineale is an Indonesian native orchid from the Spatulata section in Orchidaceae Family. This orchid is important because it is usually used as a parental plant in orchid breeding and is predicted to have a potential phytochemistry compound. In addition, in their natural habitat, this orchid is threatened due to forest exploitation and natural disaster. Therefore the precision mass propagation techniques for this orchid need to be conducted. Biotechnological approaches through inserting embryo gene such as AtRKD4 from Arabidopsis thaliana has already been successfully conducted. This study aims to check the integration stability of T-DNA harboring 35S::GR::AtRKD4 from ten selection transformants and to detect the existence of AtRKD4 protein after induction by Dexamethasone and/ Thidiazuron. The result showed that T-DNA were stably integrated into the genome of D. lineale transformants and the AtRKD4 protein with a molecular weight of 28.53 kDa was detected in D. lineale transformant plants after being induced by 15 µM DEX and 3 mgL-1 TDZ for 5 days.


Keywords


AtRKD4 gene; AtRKD4 protein; Dendrobium lineale; dexamethasone; somatic embryo; thidiazuron

Full Text:

PDF


References

Chardin, C. et al., 2014. The plant RWP-RK transcription factors: Key regulators of nitrogen responses and of gametophyte development. Journal of Experimental Botany, 65(19), pp.5577–5587.

Febryanti, N.L.P.K. et al., 2020. The expression analysis of AtRKD4 transgene in Dendrobium lineale Rolfe transgenic orchid carrying 35S::GR::AtRKD4 for micropropagation. AIP Proceeding of The 6th International Conference on Biological Science ICBS 2019: “Biodiversity as a Cornerstone for Embracing Future Humanity,” 2260 (September), 060021.

Fellers, J.P., Guenzi, A.C. & Porter, D.R., 1997. Marker proteins associated with somatic embryogenesis of wheat callus cultures. The Journal of Plant Physiology, 151, pp.201–208

Filiz, E. et al. 2018. Genetic Diversity and Phylogenetic Analyses of Turkish Rice Varieties Revealed by Issr Markers and Chloroplast Trnl-F Region. Fresenius Environ, 27(12), pp. 8351–8358.

Ghosh, A., Igamberdiev, A.U. & Debnath, S.C., 2018. Thidiazuron-induced somatic embryogenesis and changes of antioxidant properties in tissue cultures of half-high blueberry plants. Scientific Reports, 8(1), pp.1–11.

Gow, W.P. et al., 2018, ‘Factors Affecting Thidiazuron-Induced Direct Somatic Embryogenesis of Phalaenopsis aphrodite’, in Ahmad N., Faisal M. (eds.), Thidiazuron: From Urea Derivative to Plant Growth Regulator, pp.317-327, Springer, Singapore.

Hartati, S., Budiyono, A. & Cahyono, O., 2016. Pengaruh NAA dan BAP Terhadap Pertumbuhan Subkultur Anggrek Hasil Persilangan Dendrobium biggibum X Dendrobium liniale. Caraka Tani: Journal of Sustainable Agriculture, 31(1), p.33.

Heda, G.D. et al., 2016. Effects of reusing gel electrophoresis and electrotransfer buffers on western blotting. Journal of Biomolecular Techniques, 27(3), pp.113–118.

Islam, M.O., Ichihasi S., & Matsui S, 1998. Control of growth and development of protocorm like body derived from callus by carbon sources in Phalaenopsis. Plant Biotechnol, 15, pp.183-187.

Ivakdalam, L.M. & Pugesehan, D.J., 2016. Keragaman Jenis Tanaman Anggrek (Orchidaceae) Di Cagar Alam Angwarmase, Kabupaten Maluku Tenggara Barat. Jurnal Agroforestri, 11(3), pp.161-168.

Kasi, P.D. & Semiarti, E., 2016. Pengaruh Thidiazuron Dan Naphthalene Acetic Acid Untuk Induksi Embryogenesis Somatik Dari Daun Anggrek Phalaenopsis “Sogo Vivien”. Jurnal Dinamika, 7, pp.31-40.

Li, Y. et al., 1994. An Auxine Inducible Element in Soybean SAUR Promoters. Plant Physiol, 106, pp.37-43.

Mahendran, G. & Narmatha Bai, V., 2016. Direct somatic embryogenesis of Malaxis densiflora (A. Rich.) Kuntze. Journal of Genetic Engineering and Biotechnology, 14(1), pp.77–81.

Maruyama, T.E. & Hosoi, Y., 2019. Progress in somatic embryogenesis of Japanese pines. Frontiers in Plant Science, 10(January), pp.1–15.

Modi, A., & Kumar, N, 2018, ‘TDZ-induced regeneration in Stevia rebaudiana Bertoni: an important natural sweetener’, in Ahmad N., Faisal M. (eds.), Thidiazuron: From Urea Derivative to Plant Growth Regulator, pp.351- 358, Springer, Singapore.

Moradi, S. et al., 2017. Direct somatic embryogenesis in Epipactis veratrifolia, a temperate terrestrial orchid. Journal of Horticultural Science and Biotechnology, 92(1), pp.88–97. Mose, W. et al., 2017. The Influence of Thidiazuron on Direct Somatic Embryo Formation from Various Types of Explant in Phalaenopsis amabilis (L.) Blume Orchid. HAYATI Journal of Biosciences, 24(4), pp.201 –205.

Mose, W. et al., 2020. Direct Somatic Embryogenesis and Regeneration of an Indonesian orchid Phalaenopsis amabilis (L.) Blume under a Variety of Plant Growth Regulators, Light Regime, and Organic Substances. Jordan Journal of Biological Sciences, 13(4), pp.509–518.

Mursyanti, E. et al., 2015. Induction of Somatic Embryogenesis through Overexpression of ATRKD4 Genes in Phalaenopsis “Sogo Vivien.” Indonesian Journal of Biotechnology, 20(1), p.42.

Nisler, J., 2018, ‘TDZ: Mode of Action, Use and Potential in Agriculture’, in Ahmad N., Faisal M. (eds.), Thidiazuron: From Urea Derivative to Plant Growth Regulator, p.48, Springer, Singapore.

Pridgeon, A., 1992, The Illustrated Encyclopedia of Orchids, Timber Press, Inc., Oregon. Rolfe, R. A, 1889. Dendrobium lineale Rolfe. Gardeners’ Chronicle, 2, p.38.

Schena, M., Lloyd, A.M. & Davis, R.W., 1991. A steroid-inducible gene expression system for plant cells. Proceedings of the National Academy of Sciences of the United States of America, 88(23), pp.10421–10425.

Semiarti, E. et. al. 2007. Agrobacterium Mediated Transformation of The Wild Orchid Species Phalaenopsis amabilis. Plant Biotechnology, 24, pp.265- 272.

Semiarti, E. et al., 2011. Agrobacterium-Mediated Transformation of Indonesian Orchids for Micropropagation in: Alvarez, M., Genetic Transformation. Croatia: Intech, pp.215-240. Semiarti, E. et al., 2018. Stability of T-DNA Integration in Phalaenopsis “Sogo vivien” Transgenic Orchid Carrying 35S::Gal4::AtRKD4::GR. Biology Medicine & Natural Product Chemistry, 7(1), pp.5-13.

Semiarti, E., Purwantoro, A., Puspita, S.I., 2020, ‘Biotechnology Approaches on Characterization, Mass Propagation, and Breeding of Indonesian Orchids Dendrobium lineale (Rolfe.) and Vanda tricolor (Lindl.) with Its Phytochemistry’, in Merillon JM., Kodja H. (eds.), Orchids Phytochemistry, Biology and Horticulture. Reference Series in Phytochemistry. Springer, Cham.

Setiari, N. et al., 2018. Micropropagation of Dendrobium phalaenopsis orchid through overexpression of embryo gene AtRKD4. Agrivita, 40(2), pp.284–294.

Tchorbadjieva, M.I., 2005. Protein markers for somatic embryogenesis. Plant Cell Monographs, pp.215–233.

Utami, E.S.W. et al., 2007. Embriogenesis Somatik Anggrek Bulan Phalaenopsis amabilis (L.) Bl: Struktur dan Pola Perkembangan. Berkala Penelitian Hayati, 13(1), pp.33–38.

Yeşiltaş, B.N., & Kolören, O., 2019. Molecular Characterization of Sicyos Species in Ordu And Giresun Provinces. Turk. J. Weed sci, 22(1), pp.37– 44.

Zulwanis et al., 2020. The expression of AtRKD4 transgene during induction of somatic embryogenesis in transgenic dendrobium phalaenopsis Orchid Carrying 35S::GR::AtRKD4. AIP Conference Proceedings, 2260



DOI: https://doi.org/10.22146/jtbb.61783

Article Metrics

Abstract views : 99 | views : 65

Refbacks

  • There are currently no refbacks.


Copyright (c) 2021 Journal of Tropical Biodiversity and Biotechnology

Creative Commons License
This work is licensed under a Creative Commons Attribution-ShareAlike 4.0 International License.

Editoral address:

Faculty of Biology, UGM

Jl. Teknika Selatan, Sekip Utara, Yogyakarta, 55281, Indonesia

ISSN: 2540-9581 (online)