Isolation and molecular identification of agarwood-inducing fungi and their virulence test using Aquilaria sp. seedlings

##plugins.themes.bootstrap3.article.main##

LUKMAN
DINY DINARTI
ULFAH JUNIARTI SIREGAR
MAMAN TURJAMAN
SUDARSONO

Abstract


Abstract. Lukman, Dinarti D, Siregar UJ, Turjaman M, Sudarsono. 2023. Isolation and molecular identification of agarwood-inducing fungi and their virulence test using Aquilaria sp. seedlingsBiodiversitas 24: 140-148Agarwood is a lump of fragrant resin with a dark color and distinctive aroma from secondary metabolite products that accumulate in wood fibers produced from the genus Aquilaria and Gyrinops, a plant defense mechanism against damage caused by wounds and infection of specific fungal pathogens. The three main factors in producing agarwood sapwood are the presence of plants that have the potential to produce agarwood (family Thymelaceae) as a host, a supportive environment, and the presence of specific fungal pathogens. The isolation, identification, and virulence testing of local isolates must be made to get the correct agarwood-inducing pathogens without posing potential environmental problems. A total of five explants of Aquilaria sp. branches producing agarwood from the study site were used. This study aims to isolate local strains of fungi associated with agarwood formation from North Aceh regency, Aceh province, Indonesia, figure out their virulence against and ability to induce agarwood from seedlings of target trees, and determine their species identity. The experiments comprised isolation, macroscopic and microscopic identification, virulence tests, and species identification based on ITS sequences. The study resulted in five local Aceh isolates (LAI-1- LAI-5). Four isolates (LAI-1 - LAI-4) can induce agarwood formation, and oneisolate (LAI-5) is a white rot fungus. The ITS sequence analysis showed that LAI-1 - LAI-4 were Fusarium solani, while two colonies taken from LAI-5 isolate were identified as Ceriporia lacerataor Irpex laceratus (LAI-5 colony#1), and Daldinia eschscholtzii (LAI-5 colony #3). Based on these findings, the LA-5 isolate was probably a mixture of the two white rot fungus isolates.


##plugins.themes.bootstrap3.article.details##

References
Booth C. 1971. The Genus Fusarium. England: Commonwealth Mycological Institute.
Budi SWR, Santoso E & Wahyudi A. 2010. Identifikasi jenis-jenis fungi yang potensial terhadap pembentukan gaharu dari batang Aquilaria spp. J. Silvikultur Tropika. (1) :1-5. ISSN: 2086-8227
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
Daniels NM, Gallant A, Peng J, Cowen LJ, Baym M, Berger B. 2013. Compressive genomics for protein databases. Bioinformatics. 29:1283-1290.
Du TY, Dao CJ, Mapook A, Stephenson SL, Elgorban AM, Al-Rejaie S, Suwannarach N, Karunarathna SC, and Tibpromma S. 2022. Diversity and biosynthetic activities of agarwood associated fungi. Diversity 2022, 14, 211. https://doi.org/10.3390/ d14030211.
Edgar RC. 2004. MUSCLE : Multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Research, 32, 1792-1797.
Faizal A, Azar AWP, Turjaman M and. Esyanti RR. 2020. Fusarium solani induces the formation of agarwood in Gyrinops versteegii (Gilg.) Domke branches. Symbiosis 81:15–23. doi.org/10.1007/s13199-020-00677-w
Ferniah RS, Pujiyanto S, Purwantisari P dan Supriyadi. 2011. Interaksi kapang patogen Fusarium oxysporum dengan bakteri Kitinolitik rizosfer tanaman jahe dan pisang. J. Natur Indonesia 14(1): 56-60 ISSN 1410-9379 .
Gao ZH, Wei JH, Yang Y, Zhang Z, Zhao WT. 2012. Selection and validation of reference genes for studying stress-related agarwood formasion of Aquilaria sinesis. Plant Cell Rep. 31:1759?1768. doi: 10.1007/s00299-012-1289-x.
Hafizi R, Salleh B, and Latiffah Z. 2013. Morphological and molecular characterization of Fusarium solani and F. oxysporum associated with crown disease of oil palm. Brazilian Journal of Microbiology, 44(3), 959–968. https://doi.org/10.1590/S1517- 83822013000300047.
Jong PL, Tsan P, Mohamed R. 2014. Gas chromatography-mass spectrometry analysis of agarwood extracts from mature and juvenile Aquilaria malaccensis. Int J Agric Biol 16:644–648. http://www.fspublishers.org/
Legiastuti TS dan Amining T. 2012. Identifikasi cendawan endofit menggunakan teknik polymerase chain reaction. J Fitopatol Indones. ISSN: 0215-7950 31-36.
Liu Y, Chen H, Yang Y, Zhang Z, Wei J, Meng H, Wei, Meng H , Chen W, Feng J, Gan B, Chen X, Gao Z, Huang J, Chen B and Chen H.2013. Wholetree agarwood-inducing technique: an efficient novel technique for producing high-quality agarwood in cultivated Aquilaria sinensis trees. Molecules 18, 3086–3106. doi: 10.3390/molecules18033086.
Liu YY, Wei JH, Gao ZH, Zhang Z, and Lyu JC. 2017. A review of quality assessment and grading for agarwood. Chin. Herb. Med. 9, 22–30. doi: 10.1016/S1674-6384(17)60072-8.
Mohamed R, Jong PL and Kamziah AK. 2014. Fungal inoculation induces agarwood in young Aquilaria malaccensis Trees in the Nursery. J. Forestry Research, 25, 201-204. http://dx.doi.org/10.1007/s11676-013-0395-0.
Mohammad YH, Shivanand P, Metali F, Taha H, Matussinn NBA, Al-Azim-Halim AM, and Mohaimin AZ. 2021. Agarwood formation in Aquilaria beccariana and Aquilaria microcarpa in response to inoculation of newly isolated fungi from Brunei Darussalam. Biodiversitas Vol. 22, Number 10, Pages: 4131-4138. ISSN: 1412-033X, E-ISSN: 2085-4722. doi: 10.13057/biodiv/d221002
Nguyen, Tra TH, Nguyen, Duy V. 2014. Biodiversity of major bacterial groups in association with agarwood (Aquilaria crassna) in Khanh Hoa province, Vietnam. J. Viet. Env. Vol. 6, No. 2, pp. 132-137 doi: 10.13141/jve.vol6.no2. pp132-137.
Nugraha F, Roslim DI, Ardilla YP and Herman. 2014. Partial gene sequence ferritin2 on rice plants (Oryza sativa L.) Indragiri Hilir, Riau. Biosaintifika Journal of Biology & Biology Education. doi: 10.15294/biosaintifika.v6i2.3102.
Nugraheni YMMA, Anggadhania L and Putranto RA. 2015. Identification of three agarwood-forming fungi isolates from Nusa Tenggara Barat using ITS and TEF1-?. J. Pemuliaan Tanaman Hutan Vol. 9 No. 2, 77-90. doi: 10.20886/jpth.2015.9.2.77-90
Nurbaya, Tutik K., Baharuddin, Ade R. dan Syamsuddin M. 2014. Uji kecepatan pertumbuhan Fusarium spp., pada media organik dan media sintesis. J. Bionature, 15 (1) : 45-46.
Premalatha K and Kalra A. 2013. Molecular phylogenetic identification of endophytic fungi isolated from resinous and healthy wood of Aquilaria malaccensis, a red listed and highly exploited medicinal tree. Fungal Ecology 6: 205?211.
Rakhmana S, Saryono dan Nugroho TT. 2015. Ekstraksi DNA dan amplifikasi ITS rDNA isolat fungi endofit lbkurcc67 umbi tanaman dahlia (Dahlia variabilis). JOM FMIPA Vol. 2 No. 1.
Rasool S and Mohamed R. 2016. "Understanding agarwood formation and its challenges," in agarwood: Science behind the fragrance, ed. R. Mohamed. (Berlin: Springer), 39–56. doi: 10.1007/978-981-10-0833-7_3
Rayner, RW. 1970. A mycological colour chart. Mycological institute & British Mycological Society, Commonwealth England.
Sambrook J, Russell DW. 2001. Molecular Cloning a Laboratory Manual. Ed ke-3. New York: Cold Spring Harbor Laboratory.
Sangareswari M, Parthiban KT, Kanna SU, Karthiba L and Saravanakumar D. 2016. Fungal microbes associated with agarwood formation. Am. J. Plant Sci. 7, 1445–1452. http://dx.doi.org/10.4236/ajps.2016.710138.
Schoch CL, Seifert KA, Huhndorf S, Robert V, Spouge JL, Levesque CA and Schindel D. 2012. Nuclear ribosomal internal transcribed spacer (ITS) region as a universal DNA barcode marker for Fungi. In C. L. Schoch, K. A. Seifert, S. Huhndorf, V. Robert, J. L. Spouge, C. A. Levesque, W. Chen, & Fungal Barcoding Consortium (Eds.), Proceedings of the Jurnal National Academy of Sciences of the United States of America, 109, 6241-6246.
Selno S, Zakiah Z dan Kurniatuhadi R. 2021. Kualitas gaharu Aquilaria sp. dengan pemberian bioinokulan fermentasi batang pisang yang terkena penyakit layu Fusarium. J. Bios Logos Vol.11 No. 2. DOI: https://doi.org/10.35799/jbl.11.2.2021.32551.
Semangun H. 2004. Penyakit-Penyakit Tanaman Hortikultura di Indonesia. UGM Press. Yogyakarta. 2930. 850 hal.
Siburian RHS, Siregar UJ, Siregar IZ, Santoso E, Wahyudi I. 2013. Identification of Anatomical Characteristics of Aquilaria microcarpa in its Interaction with Fusarium solani. Biotropia. 20:104?111. DOI: 10.11598/btb.2013.20.2.4
Subasinghe U, and Hettiarachchi D. 2013. Agarwood resin production and resin quality of Gyrinops walla Gaertn. Int. J. Agr. Sci. 3, 357–362.
Suryawan WIGA. 2015. Pemanfaatan medium alternatif untuk pertumbuhan isolat Fusarium sp. penginduksi pembentukan gaharu pada Gyrinops versteegii (Gilg) Domke. J. Sangkareang Mataram, 1 (3) : 54-55. ISSN : 2355-9292.
Sutarman, Jalaluddin AK, Li’aini AS , and Prihatiningrum AE. 2021. Characterizations of Trichoderma sp. and its effect on ralstonia solanacearum of tobacco seedlings. J. HPT Tropika J. HPT Tropika Vol. 21, No. 1,: 8–19 ISSN: 1411-7525 E-ISSN: 2461-0399 doi : 10.23960/j.hptt.1218-19
Tan WJ, Thanh TUV, Rafael ES, Chen YS and Yeo FKS. 2021. Morphological and molecular characterization of Fusarium spp. associated with Fusarium wilt disease of Piper nigrum L. in Northwestern region of Sarawak. Malaysian J. of Microbiology, Vol 17(2), pp. 165-177: http://dx.doi.org/10.21161/mjm.200938.
Watanabe T. 2002. Pictorial atlas of soil and seed fungi morphologies of cultured fungi and key to species. CRC Press LLC. U.S.A.
Wu ZQ, Liu S, Li JF, Li MC, Du HF, Qi LK. 2017. Analysis of gene expression and quality of agarwood using Agar-bit in Aquilaria sinensis. J. Trop. For. Sci. 29, 380–388. doi: 10.26525/jtfs2017.29.3.38 0388
Ye W, Wu H, He X, Wang L, Zhang W, Li H. 2016. Transcriptome sequencing of chemically induced Aquilaria sinensis to identify genes related to agarwood formation. PLoS One 11:e0155505. doi: 10.1371/journal.pone. 0155505.

Most read articles by the same author(s)

1 2 3 4 > >>