Micropropagation of Tongkat Ali (Eurycoma Longifolia Jack) Via Somatic Embryogenesis and Direct Plant Regeneration Techniques

Eurycoma longifolia Jack or Tongkat Ali is well known among the local communities mainly for its aphrodisiac properties and its effectiveness as the cytotoxic, anti-malarial, anti-ulcer, anti-tumor promoting and anti-parasitic agent. In view of its potential commercial value as a plantation crop as...

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Main Author:	Hussein, Sobri
Format:	Thesis
Language:	English English
Published:	2004
Subjects:	Somatic embryogenesis Eurycoma longifolia Regeneration (Botany) - Techniques
Online Access:	http://psasir.upm.edu.my/id/eprint/502/1/549737_FBSB_2004_4.pdf
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id	my-upm-ir.502
record_format	uketd_dc
institution	Universiti Putra Malaysia
collection	PSAS Institutional Repository
language	English English
topic	Somatic embryogenesis Eurycoma longifolia Regeneration (Botany) - Techniques
spellingShingle	Somatic embryogenesis Eurycoma longifolia Regeneration (Botany) - Techniques Hussein, Sobri Micropropagation of Tongkat Ali (Eurycoma Longifolia Jack) Via Somatic Embryogenesis and Direct Plant Regeneration Techniques
description	Eurycoma longifolia Jack or Tongkat Ali is well known among the local communities mainly for its aphrodisiac properties and its effectiveness as the cytotoxic, anti-malarial, anti-ulcer, anti-tumor promoting and anti-parasitic agent. In view of its potential commercial value as a plantation crop as well as to conserve its germplasm, the somatic embryogenesis and direct plant regeneration of E. longifolia were carried out as these in vitro micropropagation protocols had not been reported. In attempts to establish the somatic embryos of E. longifolia, the potential of cotyledon, zygotic embryo, leaf, petiole, stem and taproot in forming embryogenic callus were examined in the basal Murashige and Skoog (MS) medium supplemented with different auxins at various concentrations. Only cotyledon explants were able to form embryogenic callus in the presence of 1.0 mg/L (w/v) of 2,4-dichlorophenoxyacetic (2,4-D) at 30%. A higher yield (60%) of embryogenic callus was obtained when the Type 4 method dissected cotyledon explants were cultured in basal MS medium containing 0.5 mg/L (w/v) of kinetin and 1.0 mg/L (w/v) of 2,4-D. The highest number of somatic embryos (45 ± 2) was observed in the same medium formulation with the addition of 1.0 g/L (w/v) activated charcoal. Subsequent transfer of these mature somatic embryos in basal MS media supplemented with 1.0 mg/L (w/v) of kinetin produced a 90% of plantlet regeneration. Addition of activated charcoal, casein hydrolysate, abscisic acid, proline and polyethylene glycol (PEG) at various concentrations into the regeneration medium did not stimulate the conversion of E. longifolia somatic embryo into plantlet. The differences between the embryogenic and non-embryogenic callus were also determined based on histological studies. Successful direct plant regeneration was obtained from the root, stem, shoot tip, axillary and adventitious bud explants. Each explant generally requires different combinations of media and plant growth regulators to produce the highest regeneration percentage. In the root explant, the best medium formulation determined was basal Juglans medium (DKW) supplemented with 1.0 mg/L (w/v) of kinetin + 1.0 mg/L (w/v) of zeatin whereas in the stem explants, basal woody plant medium (WPM) enriched with 2.0 mg/L (w/v) of BAP and 2.0 mg/L (w/v) of zeatin was found to be the best medium formulation in increasing the regeneration rate and healthy plantlets formation. Stem and root explants that were 2 cm distant from one another as has been identified as the most suitable position for attaining the maximum percentage of direct plant regeneration. Successful direct plant regeneration from in vitro and in vivo shoot tip explants of E. longifolia was achieved in basal MS medium supplemented with 3.0 and 5.0 mg/L (w/v) of kinetin, respectively. As for the direct plant regeneration from axillary bud explant, explants produced the highest regeneration capability (90%) in basal Nitsch medium (NM) supplemented with 10.0 mg/L (w/v) of zeatin while basal NM supplied with 6.0 mg/L (w/v) of zeatin produced the highest regeneration percentage in adventitious bud explants. Rooting induction of in vitro plantlets of E. longifolia was also achieved and the highest induction rate was attained in basal MS medium supplemented with 0.5 mg/L (w/v) of Indole-3-butyric acid (IBA). Acclimatization of in vitro plantlets regenerated from somatic embryogenesis and direct plant regeneration survived well with no morphological differences from the parent plants after two months of transplantation to the soil.
format	Thesis
qualification_name	Doctor of Philosophy (PhD.)
qualification_level	Doctorate
author	Hussein, Sobri
author_facet	Hussein, Sobri
author_sort	Hussein, Sobri
title	Micropropagation of Tongkat Ali (Eurycoma Longifolia Jack) Via Somatic Embryogenesis and Direct Plant Regeneration Techniques
title_short	Micropropagation of Tongkat Ali (Eurycoma Longifolia Jack) Via Somatic Embryogenesis and Direct Plant Regeneration Techniques
title_full	Micropropagation of Tongkat Ali (Eurycoma Longifolia Jack) Via Somatic Embryogenesis and Direct Plant Regeneration Techniques
title_fullStr	Micropropagation of Tongkat Ali (Eurycoma Longifolia Jack) Via Somatic Embryogenesis and Direct Plant Regeneration Techniques
title_full_unstemmed	Micropropagation of Tongkat Ali (Eurycoma Longifolia Jack) Via Somatic Embryogenesis and Direct Plant Regeneration Techniques
title_sort	micropropagation of tongkat ali (eurycoma longifolia jack) via somatic embryogenesis and direct plant regeneration techniques
granting_institution	Universiti Putra Malaysia
granting_department	Faculty of Biotechnology and Biomolecular Sciences
publishDate	2004
url	http://psasir.upm.edu.my/id/eprint/502/1/549737_FBSB_2004_4.pdf
_version_	1747810235956854784
spelling	my-upm-ir.5022013-05-27T06:48:47Z Micropropagation of Tongkat Ali (Eurycoma Longifolia Jack) Via Somatic Embryogenesis and Direct Plant Regeneration Techniques 2004-10 Hussein, Sobri Eurycoma longifolia Jack or Tongkat Ali is well known among the local communities mainly for its aphrodisiac properties and its effectiveness as the cytotoxic, anti-malarial, anti-ulcer, anti-tumor promoting and anti-parasitic agent. In view of its potential commercial value as a plantation crop as well as to conserve its germplasm, the somatic embryogenesis and direct plant regeneration of E. longifolia were carried out as these in vitro micropropagation protocols had not been reported. In attempts to establish the somatic embryos of E. longifolia, the potential of cotyledon, zygotic embryo, leaf, petiole, stem and taproot in forming embryogenic callus were examined in the basal Murashige and Skoog (MS) medium supplemented with different auxins at various concentrations. Only cotyledon explants were able to form embryogenic callus in the presence of 1.0 mg/L (w/v) of 2,4-dichlorophenoxyacetic (2,4-D) at 30%. A higher yield (60%) of embryogenic callus was obtained when the Type 4 method dissected cotyledon explants were cultured in basal MS medium containing 0.5 mg/L (w/v) of kinetin and 1.0 mg/L (w/v) of 2,4-D. The highest number of somatic embryos (45 ± 2) was observed in the same medium formulation with the addition of 1.0 g/L (w/v) activated charcoal. Subsequent transfer of these mature somatic embryos in basal MS media supplemented with 1.0 mg/L (w/v) of kinetin produced a 90% of plantlet regeneration. Addition of activated charcoal, casein hydrolysate, abscisic acid, proline and polyethylene glycol (PEG) at various concentrations into the regeneration medium did not stimulate the conversion of E. longifolia somatic embryo into plantlet. The differences between the embryogenic and non-embryogenic callus were also determined based on histological studies. Successful direct plant regeneration was obtained from the root, stem, shoot tip, axillary and adventitious bud explants. Each explant generally requires different combinations of media and plant growth regulators to produce the highest regeneration percentage. In the root explant, the best medium formulation determined was basal Juglans medium (DKW) supplemented with 1.0 mg/L (w/v) of kinetin + 1.0 mg/L (w/v) of zeatin whereas in the stem explants, basal woody plant medium (WPM) enriched with 2.0 mg/L (w/v) of BAP and 2.0 mg/L (w/v) of zeatin was found to be the best medium formulation in increasing the regeneration rate and healthy plantlets formation. Stem and root explants that were 2 cm distant from one another as has been identified as the most suitable position for attaining the maximum percentage of direct plant regeneration. Successful direct plant regeneration from in vitro and in vivo shoot tip explants of E. longifolia was achieved in basal MS medium supplemented with 3.0 and 5.0 mg/L (w/v) of kinetin, respectively. As for the direct plant regeneration from axillary bud explant, explants produced the highest regeneration capability (90%) in basal Nitsch medium (NM) supplemented with 10.0 mg/L (w/v) of zeatin while basal NM supplied with 6.0 mg/L (w/v) of zeatin produced the highest regeneration percentage in adventitious bud explants. Rooting induction of in vitro plantlets of E. longifolia was also achieved and the highest induction rate was attained in basal MS medium supplemented with 0.5 mg/L (w/v) of Indole-3-butyric acid (IBA). Acclimatization of in vitro plantlets regenerated from somatic embryogenesis and direct plant regeneration survived well with no morphological differences from the parent plants after two months of transplantation to the soil. Somatic embryogenesis Eurycoma longifolia Regeneration (Botany) - Techniques 2004-10 Thesis http://psasir.upm.edu.my/id/eprint/502/ http://psasir.upm.edu.my/id/eprint/502/1/549737_FBSB_2004_4.pdf application/pdf en public phd doctoral Universiti Putra Malaysia Somatic embryogenesis Eurycoma longifolia Regeneration (Botany) - Techniques Faculty of Biotechnology and Biomolecular Sciences English

Micropropagation of Tongkat Ali (Eurycoma Longifolia Jack) Via Somatic Embryogenesis and Direct Plant Regeneration Techniques

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