Preliminary Assessment Of Conditions Influencing Agrobacterium Rhizogenes- Mediated Transformation Of Eurycoma Longifolia Jack. Somatic Embryos
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Date
2013-12
Authors
Balakrishnan, Bhavani
Journal Title
Journal ISSN
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Publisher
Universiti Sains Malaysia
Abstract
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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Balakrishnan , Bhavani