, Volume 19, Issue 6, pp 417–423 | Cite as

Diversity of root-associated fungal endophytes in Rhododendron fortunei in subtropical forests of China

  • Chunying Zhang
  • Lijuan Yin
  • Silan Dai
Original Paper


To investigate the diversity of root endophytes in Rhododendron fortunei, fungal strains were isolated from the hair roots of plants from four habitats in subtropical forests of China. In total, 220 slow-growing fungal isolates were isolated from the hair roots of R. fortunei. The isolates were initially grouped into 17 types based on the results of internal transcribed spacer-restriction fragment length polymorphism (ITS-RFLP) analysis. ITS sequences were obtained for representative isolates from each RFLP type and compared phylogenetically with known sequences of ericoid mycorrhizal endophytes and selected ascomycetes or basidiomycetes. Based on phylogenetic analysis of the ITS sequences in GenBank, 15 RFLP types were confirmed as ascomycetes, and two as basidiomycetes; nine of these were shown to be ericoid mycorrhizal endophytes in experimental cultures. The only common endophytes of R. fortunei were identified as Oidiodendron maius at four sites, although the isolation frequency (3–65%) differed sharply according to habitat. Phialocephala fortinii strains were isolated most abundantly from two habitats which related to the more acidic soil and pine mixed forests. A number of less common mycorrhizal RFLP types were isolated from R. fortunei at three, two, or one of the sites. Most of these appeared to have strong affinities for some unidentified root endophytes from Ericaceae hosts in Australian forests. We concluded that the endophyte population isolated from R. fortunei is composed mainly of ascomycete, as well as a few basidiomycete strains. In addition, one basidiomycete strain was confirmed as a putative ericoid mycorrhizal fungus.


Rhododendron fortunei L. Fungal diversity Ericoid endophyte Ericoid mycorrhizal fungi 



We thank Prof. Runjin Liu (Institute Of Mycorrhizal Biotechnology Qingdao Agricultural University) for critical advice on the manuscript. Funding for this research was provided by Shanghai Landscape Bureau (Grand No. ZX050208).


  1. Addy HD, Hambleton S, Currah RS (2000) Distribution and molecular characterization of the root endophyte Phialocephala fortinii along an environmental gradient in the boreal forest of Alberta. Mycol Res 104:1213–1221. doi: 10.1017/S0953756200002896 CrossRefGoogle Scholar
  2. Addy HD, Piercey MM, Currah RS (2005) Microfungal endophytes in roots. Can J Bot 83:1–13. doi: 10.1139/b04-171 CrossRefGoogle Scholar
  3. Allen WK, Allaway WG, Cox GC, Valder PG (1989) Ultrastructure of mycorrhizas of Dracophyllum secundum R. Br. (Ericales: Epacridaceae). Aust J Plant Physiol 16:147–153CrossRefGoogle Scholar
  4. Allen TR, Miller T, Berch SM, Berbee ML (2003) Culturing and direct DNA extraction find different fungi from the same ericoid mycorrhizal roots. New Phytol 160:255–272. doi: 10.1046/j.1469-8137.2003.00885.x CrossRefGoogle Scholar
  5. Berch SM, Allen TR, Berbee ML (2002) Molecular detection, community structure and phylogeny of ericoid mycorrhizal fungi. Plant Soil 244:55–66. doi: 10.1023/A:1020291516884 CrossRefGoogle Scholar
  6. Bougoure DS, Cairney JWG (2005a) Assemblages of ericoid mycorrhizal and other root-associated fungi from Epacris pulchella (Ericaceae) as determined by culturing and direct DNA extraction from roots. Environ Microbiol 7:819–827. doi: 10.1111/j.1462-2920.2005.00755.x CrossRefPubMedGoogle Scholar
  7. Bougoure DS, Cairney JWG (2005b) Fungi associated with hair roots of Rhododendron lochiae (Ericaceae) in an Australian tropical cloud forest revealed by culturing and culture-independent molecular methods. Environ Microbiol 7:1743–1754. doi: 10.1111/j.1462-2920.2005.00919.x CrossRefPubMedGoogle Scholar
  8. Cairney JWG, Ashford AE (2002) Biology of mycorrhizal associations of epacrids (Ericaceae). Tansley Review No. 135. New Phytol 154:305–326. doi: 10.1046/j.1469-8137.2002.00398.x CrossRefGoogle Scholar
  9. Cairney JWG, Meharg AA (2003) Ericoid mycorrhiza: a partnership that exploits harsh edaphic conditions. Eur J Soil Sci 54:735–740. doi: 10.1046/j.1351-0754.2003.0555.x CrossRefGoogle Scholar
  10. Chambers SM, Liu G, Cairney JWG (2000) ITS rDNA sequence comparison of ericoid mycorrhizal endophytes from Woollsia pungens. Mycol Res 104:169–175. doi: 10.1017/S0953756299001306 CrossRefGoogle Scholar
  11. Gardes M, Bruns TD (1993) ITS primers with enhanced specificity for basidiomycetes—application to the identification of mycorrhizae and rusts. Mol Ecol 2:113–118. doi: 10.1111/j.1365-294X.1993.tb00005.x CrossRefPubMedGoogle Scholar
  12. Hambleton S, Currah RS (1997) Fungal endophytes from the roots of alpine and boreal Ericaceae. Can J Bot 75:1570–1581. doi: 10.1139/b97-869 CrossRefGoogle Scholar
  13. Johansson M (2001) Fungal associations of Danish Calluna vulgaris roots with special reference to ericoid mycorrhiza. Plant Soil 231:225–232. doi: 10.1023/A:1010310921738 CrossRefGoogle Scholar
  14. Jumpponen A (2001) Dark septate endophytes are they mycorrhizal? Mycorrhiza 11:207–211. doi: 10.1007/s005720100112 CrossRefGoogle Scholar
  15. Jumpponen A, Trappe JM (1998) Dark septate endophytes: a review of facultative biotrophic root-colonizing fungi. New Phytol 140:295–310. doi: 10.1046/j.1469-8137.1998.00265.x CrossRefGoogle Scholar
  16. Mclean CB, Cunnington JH, Lawrie AC (1999) Molecular diversity within and between ericoid endophytes from the Ericaceae and Epacridaceae. New Phytol 144:351–358. doi: 10.1046/j.1469-8137.1999.00510.x CrossRefGoogle Scholar
  17. Midgley DJ, Chambers SM, Cairney JWG (2004) Distribution of ericoid mycorrhizal endophytes and root-associated fungi in neighbouring Ericaceae plants in the field. Plant Soil 259:137–151. doi: 10.1023/B:PLSO.0000020947.13655.9f CrossRefGoogle Scholar
  18. Monreal M, Berch SM, Berbee M (1999) Molecular diversity of ericoid mycorrhizal fungi. Can J Bot 77:1580–1594. doi: 10.1139/cjb-77-11-1580 CrossRefGoogle Scholar
  19. Perotto S, Girlanda M, Martino E (2002) Ericoid mycorrhizal fungi: some new perspectives on old acquaintances. Plant Soil 244:41–53. doi: 10.1023/A:1020289401610 CrossRefGoogle Scholar
  20. Read DJ (1996) The structure and function of the ericoid mycorrhizal root. Ann Bot (Lond) 77:365–374. doi: 10.1006/anbo.1996.0044 CrossRefGoogle Scholar
  21. Read DJ, Kerley S (1999) The status and function of ericoid mycorrhizal systems. In: Varma A, Hock B (eds) Mycorrhiza, 2nd edn. Springer, Berlin, pp 499–520CrossRefGoogle Scholar
  22. Sharples JM, Meharg AA, Chambers SM, Cairney JWG (2000) Symbiotic solution to arsenic contamination. Nature 40:951–952CrossRefGoogle Scholar
  23. Sigler L, Allan T, Lim SR, Berch S, Berbee M (2005) Two new Cryptosporiopsis species from roots of ericaceous hosts in western North America. Stud Mycol 53:53–62CrossRefGoogle Scholar
  24. Stoyke G, Currah RS (1991) Endophytic fungi from the mycorrhizae of alpine ericoid plant. Can J Bot 69:347–352. doi: 10.1139/b91-047 CrossRefGoogle Scholar
  25. Strandberg M, Johannsson M (1999) Uptake of nutrients in Calluna vulgaris seed plants growth with and without mycorrhiza. For Ecol Manage 114:129–135. doi: 10.1016/S0378-1127(98)00387-9 CrossRefGoogle Scholar
  26. Usuki F, Abe JP, Kakishima M (2003) Diversity of ericoid mycorrhizl fungi isolated from hair roots of Rhododendron obtusum var. Kaempferi in a Japanese red pine forest. Mycoscience 44:97–102. doi: 10.1007/s10267-002-0086-8 CrossRefGoogle Scholar
  27. Xiao G, Berch SM (1992) The ericoid mycorrhizal fungi of Gaultheria shallon. Mycologia 84:470–471. doi: 10.2307/3760201 CrossRefGoogle Scholar
  28. Xiao G, Berch SM (1996) Diversity and abundance of ericoid mycorrhizal fungi to form mycorrhizal fungi of Gaultheria shallon on forest clearcut. Can J Bot 74:337–346. doi: 10.1139/b96-042 CrossRefGoogle Scholar
  29. Yu F, Zhang CY, Yin LJ, Lai ZX (2008) In vitro inoculation technology of Rhododendron fortunei L. with ericoid mycorrhizal fungi and its inoculation effect. J Fujian Agric For Univ 37(4):360–364 Natural Science EditionGoogle Scholar
  30. Zhang CY, Hou YM, Dai SL (2008) Observation on microstructure of mycorrhizal root of Rhododendron fortunei L. Acta Hortic Sin 35(11):1641–1646Google Scholar
  31. Zijlstra JD, Hof PV, Baar J, Verkley GJM, Summerbell RC, Paradi I, Braakhekke WG, Berendse F (2005) Diversity of symbiotic root endophytes of the Helotiales in ericaceous plants and the grass, Deschampsia flexuosa. Stud Mycol 53:147–162CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  1. 1.Shanghai Landscape and Gardening Research InstituteShanghaiPeople’s Republic of China
  2. 2.College of Landscape and ArchitectureBeijing Forestry UniversityBeijingPeople’s Republic of China

Personalised recommendations