Effective hygromycin concentration for selection of Agrobacterium-mediated transgenic Arabidopsis thaliana

The floral-dip transformation method for Arabidopsis thaliana has long been established. Following transformation, an important step is involving the selection for transgenic plants through antibiotics or herbicides. Hygromycin has been widely applied for transgenic plants selection. However, the...

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Main Authors:	Ee, S.F., Khairunnisa, M.B., Zeti-Azura, M.H., Noor Azmi, S., Zamri, Z.
Format:	Article
Language:	English
Published:	Penerbit Universiti Kebangsaan Malaysia 2014
Online Access:	http://journalarticle.ukm.my/7296/ http://journalarticle.ukm.my/7296/ http://journalarticle.ukm.my/7296/1/43_1_13.pdf

id	ukm-7296
recordtype	eprints
spelling	ukm-72962016-12-14T06:43:40Z http://journalarticle.ukm.my/7296/ Effective hygromycin concentration for selection of Agrobacterium-mediated transgenic Arabidopsis thaliana Ee, S.F. Khairunnisa, M.B. Zeti-Azura, M.H. Noor Azmi, S. Zamri, Z. The floral-dip transformation method for Arabidopsis thaliana has long been established. Following transformation, an important step is involving the selection for transgenic plants through antibiotics or herbicides. Hygromycin has been widely applied for transgenic plants selection. However, the concentrations used were varied between laboratories mostly in a range of 10 mg/L up to 200 mg/L. In the present study, the hygromycin sensitivity test was performed on wild-type seeds to obtain the most effective hygromycin concentration for selection of the transgenic. A standard curve on average mean of hypocotyls lengths against hygromycin concentrations was constructed. Based on hypocotyls length and leaf colour, the most effective concentration of hygromycin was determined in the range of 20 to 30 mg/L. For screening the transformed seeds, hygromycin concentration at 25 mg/L was used. Non-transgenic plants demonstrated a clear decrease in hypocotyls lengths and no root elongation, as compared to transgenic plants. The identified transgenic seedlings were further verified through polymerase chain reaction (PCR), giving fragment with an expected size of 365 bp. Penerbit Universiti Kebangsaan Malaysia 2014-06 Article PeerReviewed application/pdf en http://journalarticle.ukm.my/7296/1/43_1_13.pdf Ee, S.F. and Khairunnisa, M.B. and Zeti-Azura, M.H. and Noor Azmi, S. and Zamri, Z. (2014) Effective hygromycin concentration for selection of Agrobacterium-mediated transgenic Arabidopsis thaliana. Malaysian Applied Biology, 43 (1). pp. 119-123. ISSN 0126-8643 http://mabjournal.com/
repository_type	Digital Repository
institution_category	Local University
institution	Universiti Kebangasaan Malaysia
building	UKM Institutional Repository
collection	Online Access
language	English
description	The floral-dip transformation method for Arabidopsis thaliana has long been established. Following transformation, an important step is involving the selection for transgenic plants through antibiotics or herbicides. Hygromycin has been widely applied for transgenic plants selection. However, the concentrations used were varied between laboratories mostly in a range of 10 mg/L up to 200 mg/L. In the present study, the hygromycin sensitivity test was performed on wild-type seeds to obtain the most effective hygromycin concentration for selection of the transgenic. A standard curve on average mean of hypocotyls lengths against hygromycin concentrations was constructed. Based on hypocotyls length and leaf colour, the most effective concentration of hygromycin was determined in the range of 20 to 30 mg/L. For screening the transformed seeds, hygromycin concentration at 25 mg/L was used. Non-transgenic plants demonstrated a clear decrease in hypocotyls lengths and no root elongation, as compared to transgenic plants. The identified transgenic seedlings were further verified through polymerase chain reaction (PCR), giving fragment with an expected size of 365 bp.
format	Article
author	Ee, S.F. Khairunnisa, M.B. Zeti-Azura, M.H. Noor Azmi, S. Zamri, Z.
spellingShingle	Ee, S.F. Khairunnisa, M.B. Zeti-Azura, M.H. Noor Azmi, S. Zamri, Z. Effective hygromycin concentration for selection of Agrobacterium-mediated transgenic Arabidopsis thaliana
author_facet	Ee, S.F. Khairunnisa, M.B. Zeti-Azura, M.H. Noor Azmi, S. Zamri, Z.
author_sort	Ee, S.F.
title	Effective hygromycin concentration for selection of Agrobacterium-mediated transgenic Arabidopsis thaliana
title_short	Effective hygromycin concentration for selection of Agrobacterium-mediated transgenic Arabidopsis thaliana
title_full	Effective hygromycin concentration for selection of Agrobacterium-mediated transgenic Arabidopsis thaliana
title_fullStr	Effective hygromycin concentration for selection of Agrobacterium-mediated transgenic Arabidopsis thaliana
title_full_unstemmed	Effective hygromycin concentration for selection of Agrobacterium-mediated transgenic Arabidopsis thaliana
title_sort	effective hygromycin concentration for selection of agrobacterium-mediated transgenic arabidopsis thaliana
publisher	Penerbit Universiti Kebangsaan Malaysia
publishDate	2014
url	http://journalarticle.ukm.my/7296/ http://journalarticle.ukm.my/7296/ http://journalarticle.ukm.my/7296/1/43_1_13.pdf
first_indexed	2023-09-18T19:49:18Z
last_indexed	2023-09-18T19:49:18Z
_version_	1777406112104972288

Effective hygromycin concentration for selection of Agrobacterium-mediated transgenic Arabidopsis thaliana

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