Agrobacterium-Mediated Stable Transformation of Medicinal Plant Andrographis paniculata Callus Expressing β-glucuronidase (GUS) Gene
This study was carried out to establish a stable genetic transformation in callus culture of Andrographispaniculata mediated by Agrobacterium tumefaciens. The leaf disks of A. paniculata were infected with A. tumefaciensLBA4404 carrying a binary vector pCAMBIA1304 that contain β-glucuronidase (GUS)...
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Universitas Gadjah Mada, Yogyakarta
2013-12-01
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doaj-8752d5fc88c34ea48907af64bbfb38b32020-11-25T00:26:36ZengUniversitas Gadjah Mada, YogyakartaIndonesian Journal of Biotechnology0853-86542089-22412013-12-011829210010.22146/ijbiotech.78736589Agrobacterium-Mediated Stable Transformation of Medicinal Plant Andrographis paniculata Callus Expressing β-glucuronidase (GUS) GeneErly MarwaniAgustina TangapoFenny Martha DwivanyThis study was carried out to establish a stable genetic transformation in callus culture of Andrographispaniculata mediated by Agrobacterium tumefaciens. The leaf disks of A. paniculata were infected with A. tumefaciensLBA4404 carrying a binary vector pCAMBIA1304 that contain β-glucuronidase (GUS) and hygromycinphosphotransferase (hpt) genes. The infection was conducted by dipping method for one hour, followed byco-cultivation in the dark for three days. To examine transient GUS expression, the co-cultivated leaf disks wereassayed for β-glucuronidase activity and to obtain stable transformed callus, the co-cultivated leaf disks wereselected on the callus induction medium which contain 20 mg/l hygromycin for selection. The transformedcallus was periodically subcultured every three weeks into the fresh selection medium over the 15 weeksperiod. To test a stable transformation, the callus was subjected to PCR analysis for GUS gene detection. Theresults indicated that the co-cultivated leaf disks expressed GUS activity and proliferated to produce callus onthe selective medium. Analysis of PCR on the transformed callus indicated the presence 976 bp fragment thatconfi rmed the presence of β-glucuronidase gene. These fi ndings imply that the β-glucuronidase was stably integrated into A. paniculata callus culture.Keywords: Andrographis paniculata, Agrobacterium tumefaciens, andrographollide, transformed callus,β-glucuronidase gene.https://jurnal.ugm.ac.id/ijbiotech/article/view/7873Andrographis paniculataAgrobacterium tumefaciensandrographollidetransformed callusβ-glucuronidase gene |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Erly Marwani Agustina Tangapo Fenny Martha Dwivany |
spellingShingle |
Erly Marwani Agustina Tangapo Fenny Martha Dwivany Agrobacterium-Mediated Stable Transformation of Medicinal Plant Andrographis paniculata Callus Expressing β-glucuronidase (GUS) Gene Indonesian Journal of Biotechnology Andrographis paniculata Agrobacterium tumefaciens andrographollide transformed callus β-glucuronidase gene |
author_facet |
Erly Marwani Agustina Tangapo Fenny Martha Dwivany |
author_sort |
Erly Marwani |
title |
Agrobacterium-Mediated Stable Transformation of Medicinal Plant Andrographis paniculata Callus Expressing β-glucuronidase (GUS) Gene |
title_short |
Agrobacterium-Mediated Stable Transformation of Medicinal Plant Andrographis paniculata Callus Expressing β-glucuronidase (GUS) Gene |
title_full |
Agrobacterium-Mediated Stable Transformation of Medicinal Plant Andrographis paniculata Callus Expressing β-glucuronidase (GUS) Gene |
title_fullStr |
Agrobacterium-Mediated Stable Transformation of Medicinal Plant Andrographis paniculata Callus Expressing β-glucuronidase (GUS) Gene |
title_full_unstemmed |
Agrobacterium-Mediated Stable Transformation of Medicinal Plant Andrographis paniculata Callus Expressing β-glucuronidase (GUS) Gene |
title_sort |
agrobacterium-mediated stable transformation of medicinal plant andrographis paniculata callus expressing β-glucuronidase (gus) gene |
publisher |
Universitas Gadjah Mada, Yogyakarta |
series |
Indonesian Journal of Biotechnology |
issn |
0853-8654 2089-2241 |
publishDate |
2013-12-01 |
description |
This study was carried out to establish a stable genetic transformation in callus culture of Andrographispaniculata mediated by Agrobacterium tumefaciens. The leaf disks of A. paniculata were infected with A. tumefaciensLBA4404 carrying a binary vector pCAMBIA1304 that contain β-glucuronidase (GUS) and hygromycinphosphotransferase (hpt) genes. The infection was conducted by dipping method for one hour, followed byco-cultivation in the dark for three days. To examine transient GUS expression, the co-cultivated leaf disks wereassayed for β-glucuronidase activity and to obtain stable transformed callus, the co-cultivated leaf disks wereselected on the callus induction medium which contain 20 mg/l hygromycin for selection. The transformedcallus was periodically subcultured every three weeks into the fresh selection medium over the 15 weeksperiod. To test a stable transformation, the callus was subjected to PCR analysis for GUS gene detection. Theresults indicated that the co-cultivated leaf disks expressed GUS activity and proliferated to produce callus onthe selective medium. Analysis of PCR on the transformed callus indicated the presence 976 bp fragment thatconfi rmed the presence of β-glucuronidase gene. These fi ndings imply that the β-glucuronidase was stably integrated into A. paniculata callus culture.Keywords: Andrographis paniculata, Agrobacterium tumefaciens, andrographollide, transformed callus,β-glucuronidase gene. |
topic |
Andrographis paniculata Agrobacterium tumefaciens andrographollide transformed callus β-glucuronidase gene |
url |
https://jurnal.ugm.ac.id/ijbiotech/article/view/7873 |
work_keys_str_mv |
AT erlymarwani agrobacteriummediatedstabletransformationofmedicinalplantandrographispaniculatacallusexpressingbglucuronidasegusgene AT agustinatangapo agrobacteriummediatedstabletransformationofmedicinalplantandrographispaniculatacallusexpressingbglucuronidasegusgene AT fennymarthadwivany agrobacteriummediatedstabletransformationofmedicinalplantandrographispaniculatacallusexpressingbglucuronidasegusgene |
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