Somatic Embryos Multiplication of Eurycoma longifolia Jack in vitro by using Temporary Immersion System RITA

Hafsah, Ja'afar and Nadiawati, Alias and Dhiya Dalila, Zawawi and Noor Madihah, Mohd (2015) Somatic Embryos Multiplication of Eurycoma longifolia Jack in vitro by using Temporary Immersion System RITA. In: Proceedings of the UniSZA Research Conference 2015 (URC ’15), 14-16 April 2015, UNISZA.

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Abstract

Temporary immersion system, Recipient for Automated Temporary Immersion® (RITA® ), is one of the innovative systems that allow the production of large number of somatic embryos or plantlets in in vitro plant propagation. It has been used widely to avoid associated problems with in vitro propagation such as low multiplication rate of somatic embryos and hyperhydricity. Thus, an attempt was made to investigate the optimal parameters; immersion time and immersion frequency, for the multiplication of direct somatic embryogenesis from cotyledon culture of E. longifolia Jack by using RITA® . The basal media used for the multiplication of somatic embryogenesis was Modified Murashige and Skoog (MS) media supplemented with 0.2 mg/L indole-3-butyric acid (IBA) + 0.1 mg/L Zeatin + 0.12 mg/L thidiazuron (TDZ) + 0.1 g/L activated charcoal. Four periods of immersion time (1, 5, 10 and 15 minutes every 4 hours) were evaluated for the efficiency in somatic embryos multiplication. In order to optimize repetitive somatic embryogenesis, three different immersion frequencies (5 min immersion every 2, 4 and 8 hours) were applied. The semi solid media was also studied to be compared as control. One Way Analysis of Variance (ANOVA) showed that the highest number of secondary embryos (69.67 ± 9.73) was found significant when immersing the globular, primary embryos for 5 minutes every 4 hours as compared to other treatments tested. Apart from that, there is no significant difference for all the immersion frequencies investigated. The secondary somatic embryos obtained in this study could be further used for development of plantlet regeneration system of E. longifolia.

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