Selection of Phalaenopsis amabilis L. Blume Orchid Resistance to Hygromycin
https://doi.org/10.22146/ijbiotech.16000 Ixora Sartika Mercuriani(1), Aziz Purwantoro(2), Sukarti Moeljopawiro(3), Seonghoe Jang(4), Endang Semiarti(5*)
(1) Doctoral Program of Biotechnology, Graduate School of Universitas Gadjah Mada, Yogyakarta, Indonesia Department of Biology Education, Faculty of Mathematics and Science, Universitas Negeri Yogyakarta, Indonesia
(2) Faculty of Agriculture, Universitas Gadjah Mada, Yogyakarta, Indonesia
(3) Faculty of Biology, Universitas Gadjah Mada, Yogyakarta, Indonesia
(4) Agricultural Biotechnology Research Center,Academia Sinica Biotechnology Center Southern Taiwan, Tainan County, Taiwan, Republic of China
(5) Faculty of Biology, Universitas Gadjah Mada, Yogyakarta, Indonesia
(*) Corresponding Author
Abstract
Examination of Phalaenopsis amabilis orchid resistance to hygromycin antibiotic is an important step to do
prior to Agrobacterium-mediated genetic transformation in this orchids using Hygromycin phosphotransferase
(HPT) gene as a selection marker in the T-DNA that harboring a desired gene to be transfered. We exposed
the plant on hygromycin containing medium. The experiment was conducted using 6 weeks old P. amabilis
protocorms. These protocorms were subcultured onto NP medium supplemented with various concentration
of Hygromycin (0, 5, 10, 20, 1nd 40 mg/l). The number of survival protocorms were examined every week for
4 weeks after subcultured (WAS). The resistancy of hygromycin was calculated as ratio of death protocorms
per total protocorms). The result showed that 10 mg/l hygromycin with 1 weeks of application caused death
close to LD 50. This data indicate that P. amabilis resistance to hygromycin treatment on the appropriate
concentration 10 mg/l, and this concentration can be used for other purposes in orchid system.
prior to Agrobacterium-mediated genetic transformation in this orchids using Hygromycin phosphotransferase
(HPT) gene as a selection marker in the T-DNA that harboring a desired gene to be transfered. We exposed
the plant on hygromycin containing medium. The experiment was conducted using 6 weeks old P. amabilis
protocorms. These protocorms were subcultured onto NP medium supplemented with various concentration
of Hygromycin (0, 5, 10, 20, 1nd 40 mg/l). The number of survival protocorms were examined every week for
4 weeks after subcultured (WAS). The resistancy of hygromycin was calculated as ratio of death protocorms
per total protocorms). The result showed that 10 mg/l hygromycin with 1 weeks of application caused death
close to LD 50. This data indicate that P. amabilis resistance to hygromycin treatment on the appropriate
concentration 10 mg/l, and this concentration can be used for other purposes in orchid system.
Keywords
hygromycin; resistance; LD50; P. amabilis; protocorm
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PDF DOI: https://doi.org/10.22146/ijbiotech.16000
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