Isolation, identification and biodiversity of endophytic fungi o Thymus

Document Type : Original Research Papers


1 Department of Plant Protection, Faculty of Agriculture, Bu-Ali Sina University, Hamedan, Iran.

2 Research Center for Agriculture and Natural Resources, Hamedan, Iran.


Endophytes are considered as an important source of bioactive natural products because these
microorganisms are able to occupy unique biological niches and grow in non-ordinary
environments. Therefore, endophytic fungi of medicinal plants can be developed for medicinal
applications. On the other hand, medicinal properties of plant can be ascribed to its endophytic
fungi. Thymus sp. is widely used as a medicinal plant. In this research, the endophytic
microflora of the plant was studied. In order to isolate endophytic fungi, 800 segments of 6
Thymus species were collected from its natural habitats in Hamedan province (Iran) and
medicinal plants garden during the seasons of spring, summer and fall 2011. Finally, 95 isolates
consisting of 11 different fungal genera, 6 yeast isolates and 19 infertile mycelium were
obtained. Among the identified isolates, the most abundant genera were Alternaria, Phoma, and
Fusarium. The above mentioned genera and the genera of Cladosporium and Colletorichum
were first reported from Thymus in Iran. The genera Curvularia, Cylindrocarpon, Drecheslra,
Aspergillus, Stemphylium and Ulocladium were also, first reported from Thymus in the world.


Main Subjects

1. Li, J., Zhao, G.Z., Chen, H.H., Wang, S.Q., Zhu, W.Y., Xu, L.H., Jiang, C.L. and Li, W.J. (2008)Antitumour and antimicrobial activities of endophytic Streptomycetes from pharmaceutical plants in rainforest. Lett. Appl. Microbiol., 47, 574–580.

2. Mejía, L.C., Rojas, E.I., Maynard, Z., Van-Bael, S., Arnold, E., Hebbar, P., Samuels, G.J., Robbins, N. and Herre, E.A. (2008) Endophytic fungi as biocontrol agents of Theobroma cacao pathogens. Biol. Control., 46, 4–14.

3. Pimentel, M.R., Molina, G., Dionísio, A.P., Junior, M.R.M. and Pastore, G.M. (2011) The use of endophytes to obtain bioactive compounds and their application in biotransformation process. Biotech. Res. Int., 10.4061/2011/576286.

4. Arnold, A.E. and Herre, E.A. (2003) Canopy cover and leaf age affect colonization by tropical fungal endophytes: Ecological pattern and process in Theobroma cacao (Malvaceae). Mycologia, 95, 388-398.

5. Zhang, H.W., Song, Y.C. and Tan, R.X. (2006) Biology and chemistry of endophytes. Nat. Prod. Rep., 23, 753-771.

6. Khan, R., Shahzad, S., Choudhary, M., Shakeel, I., Khan, A. and Aqeel, A. (2010) Communities of endophytic fungi in medicinal plant Withania somnifera. Pakistan. J. Bot., 42, 1281-1287.

7. Arnold, A.E. and Engelbrecht, B.M.J. (2007) Fungal endophytes nearly double minimum leaf conductance in seedlings of a neotropical tree species. J. Trop. Ecol., 23, 369-372.

8. Khan R (2007) Isolation, identification and cultivation of endophytic fungi from medicinal plants for the production and characterization of bioactive Fungal Metabolites. PhD thesis, University of Karachi, Pakistan.

9. Leung, A.Y. and Foster, S. (1996) Encyclopaedia of common natural ingredients used in food, drugs, and cosmetics. John Wiley and Sons press, New York.

10. Dolatabadi, H.K., Mohammadi-Goltapeh, E., Moieni, A., Jaimand, K., Pakdaman-Sardrood, B. and Varma, A. (2011) Effect of Piriformospora indica and Sebacina vermifera on plant growth and essential oil yield in Thymus vulgaris in vitro and in vivo experiments. Symbiosis, 53, 29–35.

11. Naghdibadi, H. and Makkizadeh Tafti, M. (2003) Review on Thymus vulgaris L. J. Med. Plant., 7, 1-13.

12. Jamzad, Z. (2009) Thymus and Satureja species of Iran, 1st edn. Publications of Research Institute of Forests and Rangelands, Iran.

13. Fisher, P.J., Petrini, O. and Amezquita, M.M. (1992)Endophytic fungi from alpine and mediterranean species of Thymus. Nova Hedwigia, 55, 473-477.

14. Barnett, H.L. and Hunter, B.B. (1999) Illustrated genera of imperfect fungi, 4th edn. APS press. Minnesota, USA.

15. Simmons, E.G. (2007) Alternaria an identification manual. CBS Biodiversity Series.

16. Boerema, G.H., De Gruyter, J., Noordeloos, M.E. and Hamers, M.E.C. (2004) Phoma identification manual differentiation of specific and infra-specific taxa in culture. CABI publishing, United Kingdom.

17. Gerlach, W. and Nirenberg, H. (1982)The genus Fusarium - a pictorial atlas. Mitteilungen aus der Biologischen Bundesanstalt Für Land- und Forstwirtschaft, Berlin – Dahlem.

18. Nelson, P.E., Toussoun, T.A. and Marasas, W.F.O. (1983) Fusarium species, an illustrated manual foridentification.Pennsylvania State University press, America.

19. Bensch, K., Braun, U., Groenewald, J.Z. and Crous, P.W. (2012) The genus Cladosporium. Stud. Mycol., 72, 1–401.

20. Sivanesan, A. (1987) Graminicolous species of Bipolaris, Curvularia, Drechslera, Exserohillum and their teleomorphs. CAB International Mycological Institute.

21. Ellis, M.B. (1976) Dematiaceous Hyphomycetes. Commonwealth Mycological Institute, Kew, Surrey, England.

22. Suryanarayanan, T.S., Venkatesan, G. and Murali, T.S. (2003) Endophytic fungal communities in leaves of tropical forest trees: diversity and distribution patterns. Curr. Sci.,85, 489-493.

23. Petrini, O., Sieber, T.H., Toti, L. and Viret, O. (1992) Ecology, metabolite production, and substrate utilization in endophytic fungi. Nat.Toxins,1, 185–196.

24. Carroll, G.C. and Carroll, F.E. (1978) Studies on the incidence of coniferous needle endophytes in the Pacific Northwest. Can. J. Bot., 56, 3032-3043.

25. Fisher, P.J., Petrini, O. and Webster, J. (1991) Aquatic hyphomycetes and other fungi in living aquatic and terrestrial roots of Alnus glutinosa. Mycol. Res., 95, 543–547.

26. Fisher, P.J. and Petrini, O. (1990) A comparative study of fungal endophytes in xylem and bark of Alnus species in England and Switzerland. Mycol. Res., 94, 313-319.

27. Wang, Y., Guo, L.D. (2007) A comparative study of endophytic fungi in needles, bark, and xylem of Pinus tabulaeformis. Can. J. Bot., 85, 911–917.

28. Strobel, G., Daisy, B., Castillo, U. and Harper, J. (2004) Natural product fromendophytic microorganisms. J. Nat. Prod., 67, 257-268.

29. Strobel, G. and Daisy, B. (2003)Bioprospecting for microbial endophytes and their natural products. Microbiol. Mol. Biol., 67, 491-502.

30. Debbab, A., Aly, A.H., Edrada-Ebel, R.A., Müller, W.E.G., Mosaddak, M., Hakiki, A., Ebel, R. and Proksch, P. (2009a) Bioactive secondary metabolites from the endophytic fungus Chaetomium sp. isolated from Salvia officinalis growing in Morocco. Biotechnol. Agron. Soc. Environ., 13, 229-234.

31. Debbab, A., Aly, A.H., Edrada-Ebel, R.A., Wray, V., Müller, W.E.G., Totzke, F., Zirrgiebel, U., Schächtele, Ch., Kubbutat, M.H.G., Lin, W.H., Mosaddak, M., Hakiki, A.,  et al. (2009b)Bioactive metabolites from the endophytic fungus Stemphylium globuliferum isolated from Mentha pulegium. J. Nat. Prod., 72,626–631.

32. Banerjee, D., Manna, S., Mahapatra, S. and Pati, B.R. (2009) Fungal endophytes in three medicinal plants of Lamiaceae. Acta Microbiol. et Immunol. Hungarica, 56, 243-250.

33. Arnold, A.E., Maynard, Z. and Gilbert, G.S. (2001) Fungal endophytes in dicotyledonous neotropical trees: patterns of abundance and diversity. Mycol. Res., 105, 1502-1507.

34. Gange, A.C., Dey, S., Currie, A.F. and Sutton, B.C. (2007) Site- and species-specific differences in endophyte occurrence in two herbaceous plants. J. Ecol., 95, 614-622.