Edy Setiti Wida Utami and Sucipto Hariyanto and Yosephine Sri Wulan Manuhara (2018) Agrobacterium tumefaciens-mediated transformation of Dendrobium lasianthera J.J.Sm: An important medicinal orchid. Journal of Genetic Engineering and Biotechnology, 16 (2). pp. 703-709. ISSN 1687157X
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Abstract
A protocol for genetic transformation mediated by Agrobacterium tumefaciens and production of transgenic Dendrobium lasianthera has been developed for the first time. The 8-week-old protocorm explants were used as target of transformation with Agrobacterium tumefaciens strain LBA4404 carrying plasmid pG35SKNAT1. Several parameters such as infection period, Agrobacterium density, concentration of acetosyringone, and co-cultivation period were evaluated for the transformation efficiency. The data were analyzed using one-way analysis of variance (ANOVA) and Duncan’s Multiple Range Test (DMRT) with p < 0.05. Subsequently, KNAT1 gene expression was confirmed by polymerase chain reaction (PCR) analysis. The highest efficiency of transformation (70%) obtained from protocorm explants infected with Agrobacterium culture was at the OD concentration of 0.6 for 30 min, and co-cultivated with acetosyringone 100 mM for 5 days. The results of confirmation by PCR analysis show that the KNAT1 gene has been integrated and expressed in the genome of Dendrobium lasianthera transgenic. � 2018 Production and hosting by Elsevier B.V. on behalf of Academy of Scientific Research & Technology. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc- 600nd/4.0/).
| Item Type: | Article | ||||||||
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| Uncontrolled Keywords: | A. tumefaciens Dendrobium lasianthera Medicinal orchid Protocorm Transformation | ||||||||
| Subjects: | Q Science > QK Botany | ||||||||
| Divisions: | 08. Fakultas Sains dan Teknologi > Biologi | ||||||||
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| Depositing User: | Mr Akhmad Nurfizal Reza | ||||||||
| Date Deposited: | 26 Jun 2019 02:48 | ||||||||
| Last Modified: | 26 Jun 2019 02:48 | ||||||||
| URI: | http://repository.unair.ac.id/id/eprint/83512 | ||||||||
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