Skip to main content

Advertisement

Log in

Succession patterns of fungi associated to wound-induced agarwood in wild Aquilaria malaccensis revealed from quantitative PCR assay

  • Original Paper
  • Published:
World Journal of Microbiology and Biotechnology Aims and scope Submit manuscript

Abstract

Aquilaria malaccensis produces agarwood in response to wounding and fungal attack. However, information is limited regarding Aquilaria‘s interaction with its diverse fungal community. In this study, time-related changes of three natural fungal colonizers in two wounded wild A. malaccensis were tracked, beginning a few hours after wounding up to 12 months. Using species-specific primers derived from their nrITS sequences in quantitative real-time PCR (qPCR), we quantified the amount of Cunninghamella bainieri, Fusarium solani and Lasiodiplodia theobromae. Because time is a major factor affecting agarwood quantity and quality, 14 wood samples were collected at different time points, i.e., 0–18 h, 2–13 days, 2–18 weeks, and 6–12 months after wounding. qPCR data revealed that the abundance of the three species decreased over time. The fungi were detected in high numbers during the first few hours and days after wounding (40- to 25,000-fold higher levels compared with initial counts) and in low numbers (<1- to 3,200-fold higher than initially) many months later. Consistent with its role in defense response, the accumulation of secondary metabolites at the wounding site could have caused the decline in fungal abundance. Succession patterns of the two trees were not identical, indicating that fungal populations may have been affected by tree environment and wound microclimate. Our results are important for understanding the diversity of microbial community in wild Aquilaria species and their association to wound-induced agarwood formation. Fungi could be secondary triggers to agarwood production in situations where trees are wounded in attempt to induce agarwood.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  • Applied Biosystems (2011) Real-time PCR: understanding Ct. http://www3.appliedbiosystems.com/cms/groups/mcb_marketing/documents/generaldocuments/cms_053906.pdf. Accessed 26 May 2012

  • Barden A, Anak NA, Mulliken T, Song M (2000) Heart of the matter: agarwood use and trade and CITES implementation for Aquilaria malaccencis. TRAFFIC International, Cambridge

    Google Scholar 

  • Budge GE, Shaw MW, Colyer A, Pietravalle S, Boonham N (2009) Molecular tools to investigate Rhizoctonia solani distribution in soil. Plant Pathol 58:1071–1080

    Article  CAS  Google Scholar 

  • CITES (2011) Appendix II of convention on international trade in endangered species of wild fauna and flora. http://www.cites.org/eng/app/appendices.php. Accessed 26 March 2012

  • Cui J, Wang C, Guo S, Yang L, Xiao P, Wang M (2013) Evaluation of fungus-induced agilawood from Aquilaria sinensis in China. Symbiosis 60:37–44

    Article  CAS  Google Scholar 

  • Espy MJ, Uhl JR, Mitchell S, Thorvilson JN, Svien KA, Wold AD, Smith TF (2000) Diagnosis of herpes simplex virus infections in the clinical laboratory by Lightcycler PCR. J Clin Microbiol 38:795–799

    CAS  Google Scholar 

  • Gao ZH, Wei JH, Yang Y, Zhang Z, Xiong HY, Zhao WT (2012) Identification of conserved and novel microRNAs in Aquilaria sinensis based on small RNA sequencing and transcriptome sequence data. Gene 505:167–175

    Article  CAS  Google Scholar 

  • Haugland RA, Siefring SC, Wymer LJ, Brenner KP, Dufour AP (2005) Comparison of Enterococcus density measurements by quantitative polymerase chain reaction and membrane filter culture analysis at two freshwater recreational beaches. Water Res 39:559–568

    Article  CAS  Google Scholar 

  • Henson JM, French R (1993) The polymerase chain reaction and plant disease diagnosis. Annu Rev Phytopathol 31:81–109

    Article  CAS  Google Scholar 

  • 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

  • Ke D, Ménard C, Picard FJ, Boissinot M, Ouellette M, Roy PH, Bergeron MG (2000) Development of conventional and real-time PCR assays for the rapid detection of group B Streptococci. Clin Chem 46:324–331

    CAS  Google Scholar 

  • Kodsueb R, McKenzie EHC, Lumyong S, Hyde KD (2008) Fungal succession on woody litter of Magnolia liliifera (Magnoliaceae). Fungal Divers 30:55–72

    Google Scholar 

  • Lin MH, Chen TC, Kuo TT, Tseng CC, Tseng CP (2000) Real-time PCR for quantitative detection of Toxoplasma gondii. J Clin Microbiol 38:4121–4125

    CAS  Google Scholar 

  • Liu YY, Chen HQ, Yang Y, Zhang Z, Wei JH, Meng H, Chen WP, Feng JD, Gan BC, Chen XY, Gao ZH, Huang JQ, Chen B, Chen HJ (2013) Whole-tree agarwood-inducing technique: an efficient novel technique for producing high-quality agarwood in cultivated Aquilaria sinensis trees. Molecules 18:3086–3106

    Article  CAS  Google Scholar 

  • Liu X, Gong J (2012) Revealing the diversity and quantity of peritrich ciliates in environmental samples using specific primer-based PCR and quantitative PCR. Microbes Environ 27:497–503

    Article  Google Scholar 

  • Loeffler J, Henke N, Hebart H, Schmidt D, Hagmeyer L, Schumacher U, Einsele H (2000) Quantification of fungal DNA by using fluorescence resonance energy transfer and the lightcycler system. J Clin Microbiol 38:586–590

    CAS  Google Scholar 

  • Mohamed R, Jong PL, Zali MS (2010) Fungal diversity in wounded stems of Aquilaria malaccensis. Fungal Divers 43:67–74

    Article  Google Scholar 

  • Mohamed R, Jong PL, Kamziah AK (2014) Fungi inoculation induced agarwood formation in young Aquilaria malaccensis trees in the nursery. J For Res 21:201–204

    Article  Google Scholar 

  • Morrison TB, Weis JJ, Wittwer CT (1998) Quantification of low-copy transcripts by continuous SYBR Green I monitoring during amplification. Biotechniques 24:954–962

    CAS  Google Scholar 

  • Naef R (2011) The volatile and semi-volatile constituents of agarwood, the infected heartwood of Aquilaria species: a review. Flavour Fragr J 26:73–89

    Article  CAS  Google Scholar 

  • Ng LT, Chang YS, Azizol AK (1997) A review on agar (gaharu) producing Aquilaria species. J Trop For Prod 2:272–285

    Google Scholar 

  • Nitsche A, Steuer N, Schmidt CA, Landt O, Siegert W (1999) Different real-time PCR formats compared for the quantitative detection of human cytomegalovirus DNA. Clin Chem 45:1932–1937

    CAS  Google Scholar 

  • Nobuchi T, Siripatanadilok SA (2008) The formation of wood in tropical forest trees. In: Nobuchi T, Mohd Hamami S (eds) Cytological observations of Aquilaria crassna wood associated with the formation of aloeswood. UPM Press, Serdang, pp 147–160

    Google Scholar 

  • Oldfield S, Lusty C, MacKinven A (1998) The world list of threatened trees. World Conservation Press, Cambridge

    Google Scholar 

  • Pojanagaroon S, Kaewrak C (2005) Mechanical methods to stimulate aloes wood formation in Aquilaria crassna Pierre Ex H. LEC. (Kristsana) trees. In: Jatisatienr A, Paratasilpin T, Elliott S, Anusarnsunthorn V, Wedge D, Craker LE, Gardner ZE (eds) III WOCMAP congress on medicinal and aromatic plants—volume 2: conservation, cultivation and sustainable use of medicinal and aromatic plants. ISHS Acta Horticulturae, Chiang Mai

    Google Scholar 

  • Premalatha K, 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 Ecol 6:205–211

    Article  Google Scholar 

  • Promputtha I, Lumyong S, Lumyong P, McKenzie EHC, Hyde KD (2002) Fungal succession on senescent leaves of Manglietia garrettii in Doi Suthep-Pui National Park, northern Thailand. Fungal Divers 10:89–100

    Google Scholar 

  • Sivichai S, Jones EBG, Hywel-Jones N (2002) Fungal colonisation of wood in a freshwater stream at Tad Ta Phu, Khao Yai National Park, Thailand. In: Hyde KD, Jones EBG (eds) Fungal succession. Fungal Divers, vol 10, pp 113–129

  • Soehartono T, Mardiastuti A (1997) The current trade in gaharu in West Kalimatan. Biodivers Indones 1:1–10

    Google Scholar 

  • Somrithipol S, Chatmala I, Jones EBG (2002) Cirrenalia nigrospora sp nov and C tropicalis from Thailand. Nova Hedwigia 75:477–485

    Article  Google Scholar 

  • Tamuli P, Boruah P, Nath SC, Samanta R (2000) Fungi from diseased agarwood tree (Aquilaria agallocha Roxb.): two new records. Adv For Res India 22:182–187

    Google Scholar 

  • Tamuli P, Boruah P, Nath SC, Leclercq P (2005) Essential oil of eaglewood tree: a product of pathogenesis. J Essent Oil Res 17:601–604

    Article  CAS  Google Scholar 

  • Taniguchi A, Onishi H, Eguchi M (2011) Quantitative PCR assay for the detection of the parasitic ciliate Cryptocaryon irritans. Fish Sci 77:607–613

    Article  CAS  Google Scholar 

  • Wittwer CT, Herrmann MG, Moss AA, Rasmussen RP (1997) Continuous fluorescence monitoring of rapid cycle DNA amplification. Biotechniques 22:130–134

    CAS  Google Scholar 

  • Wong MT, Siah CH, Faridah QZ, Mohamed R (2013) Characterization of wound-responsive genes in Aquilaria malaccensis. J Plant Biochem Biotechnol 22(2):168–175

    Article  CAS  Google Scholar 

  • Woodhall JW, Laurenson L, Peters JC (2012) First report of Rhizoctonia solani anastomosis group 5 (AG5) in wheat in the UK. New Dis Rep 26:9

    Article  Google Scholar 

  • Zhu F, Massana R, Not F, Marie D, Vaulot D (2005) Mapping of picoeucaryotes in marine ecosystems with quantitative PCR of the 18S rRNA gene. FEMS Microbiol Ecol 52:79–92

    Article  CAS  Google Scholar 

  • Zulak KG, Bohlmann J (2010) Terpenoid biosynthesis and specialized vascular cells of conifer defense. J Integr Plant Biol 52:86–97

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by the Universiti Putra Malaysia Research University Grant Scheme (Project No. 03-03-11-1438RU). We thank the Department of Forestry, Malacca, Malaysia, for allowing us to carry out this study at the Sungai Udang Recreational Forest in Malacca.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Rozi Mohamed.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Mohamed, R., Jong, P.L. & Nurul Irdayu, I. Succession patterns of fungi associated to wound-induced agarwood in wild Aquilaria malaccensis revealed from quantitative PCR assay. World J Microbiol Biotechnol 30, 2427–2436 (2014). https://doi.org/10.1007/s11274-014-1668-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11274-014-1668-2

Keywords

Navigation