Agarwood formation in Aquilaria beccariana and Aquilaria microcarpa in response to inoculation of newly isolated fungi from Brunei Darussalam




Abstract. Mohammad YH, Shivanand P, Metali F, Taha H, Matussin NBA, Abdul-Halim AMA, Mohaimin AZ. 2021. Agarwood formation in Aquilaria beccariana and Aquilaria microcarpa in response to inoculation of newly isolated fungi from Brunei Darussalam. Biodiversitas 22: 4131-4138. Aquilaria is an important agarwood-producing tree, but naturally occurring agarwood is rare. Artificial induction method is the most commonly used to produce agarwood in a short time. Out of the 21 species of Aquilaria, Brunei Darussalam houses two species that produce high quality agarwood, namely Aquilaria beccariana and A. microcarpa. The aims of this study are to artificially induce agarwood formation using eight different induction treatments: six fungal inoculant treatments with two control treatments, and to isolate and identify fungal strains from naturally infected agarwood tree using DNA barcoding. Agarwood formation was observed at four different time intervals i.e. 1, 3, 6 and 9 months of post-inoculation. Results exhibited that the area and the total length of discoloration zone were significantly greater at 6 and 9 months of incubation period. Induction treatment did not significantly affect the area and the total length of discoloration. FTIR analysis revealed that two aromatic compounds (C – H and C – C stretching) were detected at the wavelength ranges of 3000 – 2800 cm-1, and 1500 – 1400 cm-1 respectively, and OH group was detected at 3600 – 3400 cm-1.  Nine fungal strains from Aspergillus, Fusarium, Mucor and Trichosporon were isolated and identified from a naturally infected agarwood, and the last two fungi (Mucor and Trichosporon) were never been previously reported.


Chhipa H, Kaushik N. 2017. Fungal and bacterial diversity isolated from Aquilaria malaccensis tree and soil, induces agarospirol formation within 3 months after artificial infection. Front Microbiol 8. DOI: 10.3389/fmicb.2017.01286.
Cui J, Wang C, Guo S, Yang L, Xiao P, Wang M. 2013. Evaluation of fungus-induced agilawood from Aquilaria sinensis in China. Symbiosis 60 (1): 37–44. DOI: 10.1007/s13199-013-0237-z.
Cui J, Guo S, Fu S, Xiao P, Wang M. 2013. Effects of inoculating fungi on agilawood formation in Aquilaria sinensis. Chi Sci Bull 58 (26): 3280-3287. DOI: 10.1007/s11434-013-5856-5.
Faizal A, Azar AWP, Turjaman M, Esyanti RR. 2020. Fusarium solani induces the formation of agarwood in Gyrinops versteegii (Gilg.) Domke branches. Symbiosis 81 (1): 15–23. DOI: 10.1007/s13199-020-00677-w.
Han T, Win PP. 2016. Influence of inoculation techniques on agarwood chips formation in Aquilaria malaccensis. Int J Curr Res Life Sci 5 (7): 604-607.
IUCN. 2020. The IUCN Red List of Threatened Species. Version 2021-1. Downloaded on 15 March 2021.
Jayachandaran K, Sekar I, Parthiban KT, Damodarasamy A, Suresh KK. 2014. Analysis of different grades of agarwood (Aquilaria malaccensis Lamk.) oil through GC-MS. Indian J Nat Prod Resour 5. 44- 47
Khademolhosseini R, Jafari A, Mousavi, SM, Hajfarajollah H, Noghabi KA, Manteghian M. 2019. Physicochemical characterization and optimization of glycolipid biosurfactant production by a native strain of Pseudomonas aeruginosa HAK01 and its performance evaluation for the MEOR process. RSC Advances 9 (14): 7932–7947. DOI: 10.1039/C8RA10087J.
Kumar S, Stecher G, Tamura K. 2016. MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol Biol Evol 33 (7): DOI:10.1093/molbev/mswo54.
Lee SY, Mohamed R. 2016. The origin and domestication of Aquilaria, an important agarwood-producing genus. Agarwood: 1-20. DOI: 10.1007/978-981-10-0833-7_1
Nadia S. 2012. Extraction performance study of gaharu using microwave extraction method. January.
Nasardin NRM, Hanafiah MAM, Zainon M, Ibrahim M, Rahman AIA, Baharudin ZA, Husin MHM, Mahir I, Zulkefle AA. 2018. Comparison of chemical compounds of essential oils from natural agarwood and inoculated agarwood (Roselle-based inoculation). Indones J Electr ng Comput Sci 11 (2): 677–681. DOI: 10.11591/ijeecs.v11.i2.pp677-681.
Naziz PS, Das R, Sen S. 2019. The scent of stress: Evidence from the unique fragrance of agarwood. Front Plant Sci 10. DOI: 10.3389/fpls.2019.00840.
Nobuchi T, Siripatanadilok SA. 2008. The formation of wood in tropical forest trees. In: T Nobuchi and S Mohd Hamami (eds), Cytological observations of Aquilaria crassna wood associated with the formation of aloeswood. Malaysia; UPM Press: pp. 147-160.
Nur Farhana J, Miradatul NMR, Nur Hidayah Z, Nursyuhada CH, Ahmad Fakri SM. 2014. FTIR analysis of phenolic compound as pancreatic lipase inhibitor from inoculated Aquilaria malaccensis. Malaysian J Anal Sci 18 (3): 683-689.
R Core Team (2020). A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria.
Rueden CT, Schindelin J, Hiner MC, DeZonia BE, Walter AE, Arena ET, Eliceiri KW. 2017. ImageJ2: ImageJ for the next generation of scientific image data. BMC Bioinformatics 18 (1): 1–26. DOI: 10.1186/s12859-017-1934-z.
Subasinghe SMCUP, Hettiarachchi DS, Rathnamalala E. 2012. Agarwood-type resin from Gyrinops walla Gaertn: A new discovery. J Trop For Sci 2 (2). DOI: 10.31357/jtfe.v2i2.594.
Syaima MTS, Adam F, Nizam ASM, Farah MZY, Saufi SMS. 2016. Molecular nano-sieve approach by the application of polypiperazine (PPA) based membrane for the recovery of water soluble agarwood (Aquilaria malaccensis) marker molecules. ARPN J Eng Appl Sci 11 (4): 2174-2178.
Taha H, Shivanand P, Shahminan NIN, Osman M, Abdul-Halim AMAA, Abdullah M. 2020. Isolation and identification of culturable bacteria and fungi from mixed dipterocarp and mangrove forests of Brunei Darussalam. Proc Natl Acad Sci India Section B Biol Sci 90 (3): 523–530. DOI: 10.1007/s40011-019-01119-4.
Taha H, Shivanand P, Zainudin MAA, Hadanan NA. 2021. Short communication: Identification of culturable marine fungi and bacteria from coastal region in Brunei Darussalam. Biodiversitas 22 (3): 1326–1331. DOI: 10.13057/biodiv/d220332.
Wong MT, Siah CH, Faridah QZ, Mohamed R. 2013. Characterization of wound responsive genes in Aquilaria malaccensis. J Plant Biochem Biot 22 (2): 168–175. DOI: 10.1007/s13562-012-0144-z.
Xu Y, Zhang Z, Wang M, Wei J, Chen H, Gao Z, Sui C, Luo H, Zhang X, Yang Y, Meng H, Li W. 2013. Identification of genes related to agarwood formation: Transcriptome analysis of healthy and wounded tissues of Aquilaria sinensis. BMC Genomics 14 (1): DOI: 10.1186/1471-2164-14-227.
Zhang Z, Wei J, Han X, Liang L, Yang Y, Meng H. 2014. The sesquiterpene biosynthesis and vessel-occlusion formation in stems of Aquilaria sinensis ( Lour .) Gilg Trees induced by wounding treatments without variation of microbial communities. Int J Mol Sci 15 (12): 23589–23603. DOI: 10.3390/ijms151223589.