Preliminary Assessment Of Conditions Influencing Agrobacterium Rhizogenes- Mediated Transformation Of Eurycoma Longifolia Jack. Somatic Embryos

Effective propagation of in vitro callus, shoot, plantlet and somatic embryo cultures of Eurycoma longifolia was achieved using micropropagation technique. In vitro shoots, somatic embryos and plantlet cultures were used for conducting histological and SEM analysis, chemotaxis assays and transformat...

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Main Author: Balakrishnan, Bhavani
Format: Thesis
Language:English
Published: 2013
Subjects:
Online Access:http://eprints.usm.my/46057/
http://eprints.usm.my/46057/1/Bhavani%20Balakrishnan.pdf
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author Balakrishnan, Bhavani
author_facet Balakrishnan, Bhavani
author_sort Balakrishnan, Bhavani
building USM Institutional Repository
collection Online Access
description Effective propagation of in vitro callus, shoot, plantlet and somatic embryo cultures of Eurycoma longifolia was achieved using micropropagation technique. In vitro shoots, somatic embryos and plantlet cultures were used for conducting histological and SEM analysis, chemotaxis assays and transformation with four different Agrobacterium rhizogenes disarmed strains. Histological analysis and SEM observations revealed that somatic embryos of Eurycoma longifolia share similar traits with zygotic embryos. Positive chemotactic responses of A. rhizogenes towards somatic embryo explants were detected using swarm plate assay. In vitro shoot and plantlet cultures were used to obtain leaves, stems and rachis explants for transformation. Hairy root-like structures were seen emerging from in vitro stem explants co-cultivated with Agrobacterium rhizogenes strain AR14. However, the number of explants expressing these hairy root-like structures was too low to be confirmed using GUS assay or PCR based detection of transgenes. Transformation optimisations with somatic embryo explants were conducted by analysing the effect of four different factors on the GUS positive frequency. It was ascertained that AR12 is the most virulent and suitable Agrobacterium rhizogenes strain for transformation of E. longifolia somatic embryos.
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institution Universiti Sains Malaysia
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language English
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publishDate 2013
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spelling usm-460572020-01-31T02:16:03Z http://eprints.usm.my/46057/ Preliminary Assessment Of Conditions Influencing Agrobacterium Rhizogenes- Mediated Transformation Of Eurycoma Longifolia Jack. Somatic Embryos Balakrishnan, Bhavani QH1 Natural history (General - Including nature conservation, geographical distribution) Effective propagation of in vitro callus, shoot, plantlet and somatic embryo cultures of Eurycoma longifolia was achieved using micropropagation technique. In vitro shoots, somatic embryos and plantlet cultures were used for conducting histological and SEM analysis, chemotaxis assays and transformation with four different Agrobacterium rhizogenes disarmed strains. Histological analysis and SEM observations revealed that somatic embryos of Eurycoma longifolia share similar traits with zygotic embryos. Positive chemotactic responses of A. rhizogenes towards somatic embryo explants were detected using swarm plate assay. In vitro shoot and plantlet cultures were used to obtain leaves, stems and rachis explants for transformation. Hairy root-like structures were seen emerging from in vitro stem explants co-cultivated with Agrobacterium rhizogenes strain AR14. However, the number of explants expressing these hairy root-like structures was too low to be confirmed using GUS assay or PCR based detection of transgenes. Transformation optimisations with somatic embryo explants were conducted by analysing the effect of four different factors on the GUS positive frequency. It was ascertained that AR12 is the most virulent and suitable Agrobacterium rhizogenes strain for transformation of E. longifolia somatic embryos. 2013-12 Thesis NonPeerReviewed application/pdf en http://eprints.usm.my/46057/1/Bhavani%20Balakrishnan.pdf Balakrishnan, Bhavani (2013) Preliminary Assessment Of Conditions Influencing Agrobacterium Rhizogenes- Mediated Transformation Of Eurycoma Longifolia Jack. Somatic Embryos. Masters thesis, Universiti Sains Malaysia.
spellingShingle QH1 Natural history (General - Including nature conservation, geographical distribution)
Balakrishnan, Bhavani
Preliminary Assessment Of Conditions Influencing Agrobacterium Rhizogenes- Mediated Transformation Of Eurycoma Longifolia Jack. Somatic Embryos
title Preliminary Assessment Of Conditions Influencing Agrobacterium Rhizogenes- Mediated Transformation Of Eurycoma Longifolia Jack. Somatic Embryos
title_full Preliminary Assessment Of Conditions Influencing Agrobacterium Rhizogenes- Mediated Transformation Of Eurycoma Longifolia Jack. Somatic Embryos
title_fullStr Preliminary Assessment Of Conditions Influencing Agrobacterium Rhizogenes- Mediated Transformation Of Eurycoma Longifolia Jack. Somatic Embryos
title_full_unstemmed Preliminary Assessment Of Conditions Influencing Agrobacterium Rhizogenes- Mediated Transformation Of Eurycoma Longifolia Jack. Somatic Embryos
title_short Preliminary Assessment Of Conditions Influencing Agrobacterium Rhizogenes- Mediated Transformation Of Eurycoma Longifolia Jack. Somatic Embryos
title_sort preliminary assessment of conditions influencing agrobacterium rhizogenes- mediated transformation of eurycoma longifolia jack. somatic embryos
topic QH1 Natural history (General - Including nature conservation, geographical distribution)
url http://eprints.usm.my/46057/
http://eprints.usm.my/46057/1/Bhavani%20Balakrishnan.pdf