Agarwood induction in cell suspension culture of Aquilaria malaccensis Lam.

Most species of Aquilaria in the Thymelaeaceae family produce agarwood or ‘gaharu’. In Malaysia, the primary producer of agarwood is Aquilaria malaccensis, a tree species locally known as ‘karas’. Agarwood is one of the precious woods on earth and prized for its rich and wonderful fragrance. This st...

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Main Author:	Jayaraman, Shashita
Format:	Thesis
Language:	English
Published:	2013
Subjects:	Agarwood (Wood) Thymelaeaceae
Online Access:	http://psasir.upm.edu.my/id/eprint/39628/1/FH%202013%209%20IR.pdf
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id	my-upm-ir.39628
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spelling	my-upm-ir.396282016-04-20T07:29:03Z Agarwood induction in cell suspension culture of Aquilaria malaccensis Lam. 2013-11 Jayaraman, Shashita Most species of Aquilaria in the Thymelaeaceae family produce agarwood or ‘gaharu’. In Malaysia, the primary producer of agarwood is Aquilaria malaccensis, a tree species locally known as ‘karas’. Agarwood is one of the precious woods on earth and prized for its rich and wonderful fragrance. This study aimed at inducing agarwood production in cell suspension culture of Aquilaria malaccensis, by investigating the optimal growth medium, optimal inocula size and finding suitable elicitors. To establish cell suspension culture, callus was first induced from nursery-derived leaf explants incubated on basic Murashige and Skoog (MS) solid medium containing 1.5% sucrose, at pH 5.7. Different auxin types including 1-naphthaleneacetic acid (NAA), 2,4-dichlorophenoxyacetic acid (2,4-D), and indole-3-butyric acid (IBA), were tested at various concentrations (0.55, 1.1 and 1.65 μM). Compact type callus was induced by 1.1 μM NAA with highest biomass dry weight (DW) of 17.3 mg. This auxin concentration was then combined with either 6-benzylaminopurine (BAP) or kinetin, at 0.55, 1.1, 2.2 and 3.3 μM, to induce growth of friable callus. The 1.1μM NAA + 2.2μM BAP combination produced friable callus with the highest biomass (93.3mg DW). Microscopic observations revealed the arrangement of the friable callus as loosely packed with relatively large cells. However, for the compact callus, the cells were small and densely packed. Further investigation on effects of elicitor in agarwood production were studied using cell suspension culture with initial an inocula of 4% (fresh weight, FW) of callus. The culture was challenged with fungal elicitor prepared from two species of fungi, Trichoderma and Lasiodiplodia. Fungal elicitor in the form of mycelial crude extract was added to the medium to a final concentration of 2, 4, 6, 8, and 10 mg/L, respectively. A light scent of agarwood was detected from the culture after it had been challenged for 20 days with 8mg/L Trichoderma extract. To increase scent intensity, cell suspension cultures were initiated from 4%, 8%, 16% and 32% (FW) of calli and elicited with 8 and 16 mg/L Trichoderma extract, respectively. The combination of 16% of inocula and 8 mg/L Trichoderma extract produced the most intense fragrance comparable to agarwood scent. Several important agarwood compounds were detected using GC-MS including 8-epi-.gama.-eudesmol, á-guaiene, alloaromadendrene oxide-1 and chromone,5-hydroxy-6,7,8-trimethoxy-2,3-dimethyl. Trichoderma appeared to be a suitable elicitor for agarwood production. In conclusion, the presence of these compounds was evidence that agarwood had been induced in the fungal inoculated cell suspension culture. Agarwood (Wood) Thymelaeaceae 2013-11 Thesis http://psasir.upm.edu.my/id/eprint/39628/ http://psasir.upm.edu.my/id/eprint/39628/1/FH%202013%209%20IR.pdf application/pdf en public masters Universiti Putra Malaysia Agarwood (Wood) Thymelaeaceae
institution	Universiti Putra Malaysia
collection	PSAS Institutional Repository
language	English
topic	Agarwood (Wood) Thymelaeaceae
spellingShingle	Agarwood (Wood) Thymelaeaceae Jayaraman, Shashita Agarwood induction in cell suspension culture of Aquilaria malaccensis Lam.
description	Most species of Aquilaria in the Thymelaeaceae family produce agarwood or ‘gaharu’. In Malaysia, the primary producer of agarwood is Aquilaria malaccensis, a tree species locally known as ‘karas’. Agarwood is one of the precious woods on earth and prized for its rich and wonderful fragrance. This study aimed at inducing agarwood production in cell suspension culture of Aquilaria malaccensis, by investigating the optimal growth medium, optimal inocula size and finding suitable elicitors. To establish cell suspension culture, callus was first induced from nursery-derived leaf explants incubated on basic Murashige and Skoog (MS) solid medium containing 1.5% sucrose, at pH 5.7. Different auxin types including 1-naphthaleneacetic acid (NAA), 2,4-dichlorophenoxyacetic acid (2,4-D), and indole-3-butyric acid (IBA), were tested at various concentrations (0.55, 1.1 and 1.65 μM). Compact type callus was induced by 1.1 μM NAA with highest biomass dry weight (DW) of 17.3 mg. This auxin concentration was then combined with either 6-benzylaminopurine (BAP) or kinetin, at 0.55, 1.1, 2.2 and 3.3 μM, to induce growth of friable callus. The 1.1μM NAA + 2.2μM BAP combination produced friable callus with the highest biomass (93.3mg DW). Microscopic observations revealed the arrangement of the friable callus as loosely packed with relatively large cells. However, for the compact callus, the cells were small and densely packed. Further investigation on effects of elicitor in agarwood production were studied using cell suspension culture with initial an inocula of 4% (fresh weight, FW) of callus. The culture was challenged with fungal elicitor prepared from two species of fungi, Trichoderma and Lasiodiplodia. Fungal elicitor in the form of mycelial crude extract was added to the medium to a final concentration of 2, 4, 6, 8, and 10 mg/L, respectively. A light scent of agarwood was detected from the culture after it had been challenged for 20 days with 8mg/L Trichoderma extract. To increase scent intensity, cell suspension cultures were initiated from 4%, 8%, 16% and 32% (FW) of calli and elicited with 8 and 16 mg/L Trichoderma extract, respectively. The combination of 16% of inocula and 8 mg/L Trichoderma extract produced the most intense fragrance comparable to agarwood scent. Several important agarwood compounds were detected using GC-MS including 8-epi-.gama.-eudesmol, á-guaiene, alloaromadendrene oxide-1 and chromone,5-hydroxy-6,7,8-trimethoxy-2,3-dimethyl. Trichoderma appeared to be a suitable elicitor for agarwood production. In conclusion, the presence of these compounds was evidence that agarwood had been induced in the fungal inoculated cell suspension culture.
format	Thesis
qualification_level	Master's degree
author	Jayaraman, Shashita
author_facet	Jayaraman, Shashita
author_sort	Jayaraman, Shashita
title	Agarwood induction in cell suspension culture of Aquilaria malaccensis Lam.
title_short	Agarwood induction in cell suspension culture of Aquilaria malaccensis Lam.
title_full	Agarwood induction in cell suspension culture of Aquilaria malaccensis Lam.
title_fullStr	Agarwood induction in cell suspension culture of Aquilaria malaccensis Lam.
title_full_unstemmed	Agarwood induction in cell suspension culture of Aquilaria malaccensis Lam.
title_sort	agarwood induction in cell suspension culture of aquilaria malaccensis lam.
granting_institution	Universiti Putra Malaysia
publishDate	2013
url	http://psasir.upm.edu.my/id/eprint/39628/1/FH%202013%209%20IR.pdf
_version_	1747811798067707904

Agarwood induction in cell suspension culture of Aquilaria malaccensis Lam.

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