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All Journal HAYATI Journal of Biosciences Proceedings of Annual International Conference Syiah Kuala University - Life Sciences & Engineering Chapter
CHAN LAI KENG
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Agriculture, Biological Sciences & Forestry Earth & Planetary Sciences Engineering Other

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Effects of Medium Constituents on Growth and Canthinone Accumulation in Cell Suspension Cultures of Eurycoma longifolia Jack LUTHFI AZIZ MAHMUD SIREGAR; CHAN LAI KENG; BOEY PENG LIM
HAYATI Journal of Biosciences Vol. 16 No. 2 (2009): June 2009
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (525.15 KB) | DOI: 10.4308/hjb.16.2.69

Abstract

The effect of various macronutrients, micronutrients and sucrose on growth and canthinone alkaloid production in cell suspension cultures of Pasak Bumi (Eurycoma longifolia Jack) was investigated. The optimum macronutrients and micronutrients content for the high alkaloid production of E. longifolia Jack was different to that found in the Murashige and Skoog (MS) medium. The highest amount of alkaloids, 9-hydroxycanthin-6-one and 9-methoxycanthin-6-one, could be obtained from E. longifolia Jack cells cultured in modified MS liquid medium that containing macronutrients: 21.50 mM NH4NO3, 14.25 mM KNO3, 7.50 mM CaCl2•2H2O, 2.50 mM MgSO4•7H2O, 1.45 mM KH2PO4, while content of micronutrients was 0.233 mM FeNa-EDTA, 0.215 mM MnSO4•4H2O and without CuSO4•5H2O. Increased sucrose concentration to 4.00% (w/v) in modified MS liquid medium could increase total of two-alkaloid. The modification of macronutrients and micronutrients concentration based the optimum production of biomass was obtained MSBs medium that producing high biomass but also increasing the production of 9-hydroxycanthin-6-one. The modification of macronutrients or macronutrients and micronutrients based the optimum total of two-alkaloid was obtained MSC and MSD medium that producing low fresh weight but producing the high 9-hydroxycanthin-6-one. Key words: Pasak Bumi, 9-hydroxycanthin-6-one, 9-methoxycanthin-6-one, macronutrients, micronutrients, sucrose
Regeneration rates of Dendrobium bobby messina plbs with ascorbic acid using PVS2 vitrification Jessica Jeyanthi James Antony; Chan Lai Keng; Sreeramanan Subramaniam
Proceedings of The Annual International Conference, Syiah Kuala University - Life Sciences & Engineering Chapter Vol 2, No 1 (2012): Life Sciences
Publisher : Syiah Kuala University

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Cryopreservation techniques using PVS2 vitrification was applied on PLBs of Dendrobium Bobby Messina, with survival monitored through observations of growth rate and the 2,3,5-triphenyltetrazolium chloride (TTC) analyses. The parameters optimized were PLBs size, preculture concentration, preculture duration, PVS2 incubation temperature and duration. The optimized parameters obtained were 3-4mm of PLBs precultured in 0.2M sucrose for 1 day, treated with a mixture of 2M glycerol and 0.4M sucrose supplemented with half strength liquid MS media at 25°C for 20 minutes and subsequently dehydrated with plant vitrification solution 2 (PVS2) at 0°C for 20 minutes prior storage in liquid nitrogen. Following rapid warming in a water bath at 40°C for 90 seconds, PLBs were washed with a half strength liquid MS media supplemented with 1.2M sucrose. Subsequently, PLBs were cultured on half strength semi-solid MS media supplemented with 2% (w/v) sucrose in the absence of growth regulator. The optimized vitrification method was successful in preserving this orchid as it produced growth recovery in cryopreserved PLBs up to 40%. Ascorbic acid was added in the media to evaluate the regeneration process of cryopreserved PLBs. However, growth recovery rate was only 10% at 0.6mM ascorbic acid. RAPD analysis using 6 primers indicated that cryopreserved and non-cryopreserved PLBs from vitrification method were genetically faithful to the mother plant. However, 3 primers showed polymorphism and 1 primers indicated partial polymorphism between the cryopreserved and non-cryopreserved PLBs in comparative to the mother plant.
The effect of strain virulence on Agrobacterium rhizogenes transformation efficiency in Eurycoma longifolia Bhavani Balakrishnan; Advina Lizah Julkifle; Syarifah Shahrul Rabiah Syed Alwee; Chan Lai Keng; Sreeramanan Subramaniam
Proceedings of The Annual International Conference, Syiah Kuala University - Life Sciences & Engineering Chapter Vol 2, No 1 (2012): Life Sciences
Publisher : Syiah Kuala University

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (113.207 KB)

Abstract

Eurycoma longifolia, known locally as ‘Tongkat Ali’ in Malaysia, is popularly sought out as herbal remedy in many parts of South East Asia. Consequently, this has resulted in the indiscriminate collection of the taproot from the wild, leading to fears of extinction among conservationists. Agrobacterium rhizogenes is a naturally occurring soil bacterium that infects injured plants and causes a massive proliferation of roots, known as hairy roots. The objective of this study is to determine the transformation efficiency of 2 different strains of Agrobacterium rhizogenes on somatic embryos of Eurycoma longifolia using transient GUS expression as an indicator of successful transformation. Somatic embryos cultured in vitro for 4 to 5 weeks were used as explants and were pre-cultured for 2 days in full strength MS medium containing 0.5 mg/L IBA added with 1% PVP and 2mg/L DTT. The explants were transformed using Agrobacterium rhizogenes strains AR12 and AR14. GUS assay was carried out 1 week after transformation and observed. Observations indicate that AR12 is more effective in transforming somatic embryos of Eurycoma longifolia compared to AR14. Therefore, Agrobacterium rhizogenes strain AR12 is a potential candidate for the successful transformation of Eurycoma longifolia somatic embryos, leading to the induction of sustainable hairy root cultures
Co-Authors Advina Lizah Julkifle Bhavani Balakrishnan BOEY PENG LIM Jessica Jeyanthi James Antony Luthfi Aziz Mahmud Siregar Sreeramanan Subramaniam Sreeramanan Subramaniam Syarifah Shahrul Rabiah Syed Alwee