Antonie van Leeuwenhoek

, Volume 107, Issue 6, pp 1451–1473 | Cite as

New Ceratocystis species from Eucalyptus and Cunninghamia in South China

  • FeiFei Liu
  • Michael Mbenoun
  • Irene Barnes
  • Jolanda Roux
  • Michael J. Wingfield
  • GuoQing Li
  • JieQiong Li
  • ShuaiFei ChenEmail author
Original Paper


During routine surveys for possible fungal pathogens in the rapidly expanding plantations of Eucalyptus and Cunninghamia lanceolata in China, numerous isolates of unknown species in the genus Ceratocystis (Microascales) were obtained from tree wounds. In this study we identified the Ceratocystis isolates from Eucalyptus and Cunninghamia in the GuangDong, GuangXi, FuJian and HaiNan Provinces of South China based on morphology and through comparisons of DNA sequence data for the ITS, partial β-tubulin and TEF-1α gene regions. Morphological and DNA sequence comparisons revealed two previously unknown species residing in the Indo-Pacific Clade. These are described here as Ceratocystis cercfabiensis sp. nov. and Ceratocystis collisensis sp. nov. Isolates of Ceratocystis cercfabiensis showed intragenomic variation in their ITS sequences and four strains were selected for cloning of the ITS gene region. Twelve ITS haplotypes were obtained from 17 clones selected for sequencing, differing in up to seven base positions and representing two separate phylogenetic groups. This is the first evidence of multiple ITS types in isolates of Ceratocystis residing in the Indo-Pacific Clade. Caution should thus be exercised when using the ITS gene region as a barcoding marker for Ceratocystis species in this clade. This study also represents the first record of a species of Ceratocystis from Cunninghamia.


Ceratocystidaceae Fungal barcoding genes Multiple ITS types Plantation forestry 



This study was initiated through the bilateral agreement between the Governments of South Africa and China, and we are grateful for the funding via projects 2012DFG31830 (International Science & Technology Cooperation Program of China), 31400546 (National Natural Science Foundation of China: NSFC), 2010KJCX015-03 (Forestry Science and Technology Innovation Project of Guangdong Province of China). We acknowledge members of Tree Protection and Cooperation Programme (TPCP) and the National Research Foundation (NRF), South Africa for financial support. Tao Huang is thanked for assistance with the fieldwork, and Arista Fourie for the help with the cloning of isolates.


  1. Al Adawi AO, Barnes I, Khan IA, Al Subhi AM, Al Jahwari AA, Deadman ML, Wingfield BD, Wingfield MJ (2013) Ceratocystis manginecans associated with a serious wilt disease of two native legume trees in Oman and Pakistan. Australas Plant Pathol 42:179–193CrossRefGoogle Scholar
  2. Barnes I, Roux J, Wingfield BD, Neill MO, Wingfield MJ (2003) Ceratocystis fimbriata infecting Eucalyptus grandis in Uruguay. Australas Plant Pathol 32:361–366CrossRefGoogle Scholar
  3. Camille F, Morrell JJ (2006) Decay resistance of China-fir (Cunninghamia lanceolata (Lambert) Hooker). For Prod J 56:29–30Google Scholar
  4. Chen SF, van Wyk M, Roux J, Wingfield MJ, Xie YJ, Zhou XD (2013) Taxonomy and pathogenicity of Ceratocystis species on Eucalyptus trees in South China, including C. Chinaeucensis sp. nov. Fungal Divers 58:267–279CrossRefGoogle Scholar
  5. Cunningham CW (1997) Can three incongruence tests predict when data should be combined. Mol Biol Evol 14:733–740CrossRefPubMedGoogle Scholar
  6. De Beer ZW, Duong TA, Barnes I, Wingfield BD, Wingfield MJ (2014) Redefining Ceratocystis and allied genera. Stud Mycol 79:187–219CrossRefPubMedCentralPubMedGoogle Scholar
  7. Engelbrecht CJ, Harrington TC, Alfenas A (2007) Ceratocystis wilt of cacao—a disease of increasing importance. Phytopathology 97:1648–1649CrossRefPubMedGoogle Scholar
  8. Felsenstein J (1985) Confidence intervals on phylogenetics: an approach using bootstrap. Evolution 39:783–791CrossRefGoogle Scholar
  9. Fourie A, Wingfield MJ, Wingfield BD, Barnes I (2015) Molecular markers delimit cryptic species in Ceratocystis sensu stricto. Mycol Prog 14:1–18CrossRefGoogle Scholar
  10. Glass NL, Donaldson GC (1995) Development of primer sets designed for use with the PCR to amplify conserved genes from filamentous Ascomycetes. Appl Environ Microb 6:1323–1330Google Scholar
  11. Guindon S, Gascuel O (2003) A simple, fast, and accurate algorithm to estimate large phylogenies by maximum likelihood. Syst Biol 52:696–704CrossRefPubMedGoogle Scholar
  12. Halsted BD (1890) Some fungous disease of sweet potato. N J Agric Coll Exp Stn Bull 76:1–32Google Scholar
  13. Harrington TC (2004) CABI crop protection compendium. Cabi publishing http://www.public.
  14. Harrington TC, Kazmi MR, Al Sadi AM, Ismail SI (2014) Intraspecific and intragenomic variability of ITS rDNA sequences reveals taxonomic problems in Ceratocystis fimbriata sensu stricto. Mycologia 106:224–242CrossRefPubMedGoogle Scholar
  15. Hausner G, Reid J, Klassen GR (1993) On the phylogeny of Ophiostoma, Ceratocystis s.s., and Microascus, and relationships within Ophiostoma based on partial ribosomal DNA sequences. Can J Bot 71:1249–1265CrossRefGoogle Scholar
  16. Hillis DM, Huelsenbeck JP (1992) Signal, noise and reliability in molecular phylogenetic analyses. J Hered 83:189–195PubMedGoogle Scholar
  17. Huang Q, Zhu YY, Chen HR, Wang YY, Liu YL, Lu WJ, Ruan XY (2003) First report of pomegranate wilt caused by Ceratocystis fimbriata in Yunnan, China. Plant Dis 87:1150CrossRefGoogle Scholar
  18. Huang Q, Wang YY, Zhao YY, Jiao YX, Li XF, Chen HR, Zhu YY (2008) First report of taro black rot caused by Ceratocystis fimbriata in China. Plant Pathol 57:780CrossRefGoogle Scholar
  19. Jacobs K, Bergdahl DR, Wingfield MJ, Halik S, Seifert KA, Bright DE, Wingfield BD (2004) Leptographium wingfieldii introduced into North America and found associated with exotic Tomicus piniperda and native bark beetles. Mycol Res 108:411–418CrossRefPubMedGoogle Scholar
  20. Johnson JA, Harrington TC, Engelbrecht CJB (2005) Phylogeny and taxonomy of the North American clade of the Ceratocystis fimbriata complex. Mycologia 97:1067–1092CrossRefPubMedGoogle Scholar
  21. Katoh K, Misawa K, Kuma K, Miyata T (2002) MAFFT: a novel method for rapid multiple sequence alignment based on fast Fourier transform. Nucleic Acids Res 30:3059–3066CrossRefPubMedCentralPubMedGoogle Scholar
  22. Kile GA (1993) Plant diseases caused by species of Ceratocystis sensu strict and Chalara. In: Wingfield MJ, Seifert KA, Webber JFF (eds) Ceratocystis and Ophiostoma: taxonomy, ecology and pathogenicity. American Phytopathological Society Press, St Paul, pp 73–183Google Scholar
  23. Li J, Gao JM, Han YH, Sun YX, Huang Q (2014a) First report of Ceratocystis fimbriata-caused wilt of Eriobotrya japonica in China. Plant Dis 98:1270CrossRefGoogle Scholar
  24. Li J, Zhang Y, Xu KC, Yang JY, Han YH, Sun YX, Huang Q (2014b) First report of wilt of Eucalyptus caused by Ceratocystis fimbriata in China. Plant Dis. doi: 10.1094/PDIS-06-14-0580-PDN Google Scholar
  25. Liu L, Duan ZH, Xu MK et al (2010) Effect of monospecific and mixed Cunninghamia lanceolata plantations on microbial community and two functional genes involved in nitrogen cycling. Plant Soil 327:413–428CrossRefGoogle Scholar
  26. Mbenoun M, Wingfield MJ, Aimé D, Boyogueno B, Wingfield BD, Roux J (2014) Molecular phylogenetic analyses reveal three new Ceratocystis species and provide evidence for geographic differentiation of the genus in Africa. Mycol Prog 13:219–240CrossRefGoogle Scholar
  27. Möller EM, Bahnweg G, Sandermann H, Geiger HH (1992) A simple and efficient protocol for isolation of high molecular weight DNA from filamentous fungi, fruit bodies, and infected plant tissues. Nucleic Acids Res 20:6115–6116CrossRefPubMedCentralPubMedGoogle Scholar
  28. Morris MJ, Wingfield MJ, de Beer C (1993) Gummosis and wilt of Acasia mearnsii in South Africa caused by Ceratocystis fimbriata. Plant Pathol 42:814–817CrossRefGoogle Scholar
  29. Naidoo K, Steenkamp ET, Coetzee MPA, Wingfield MJ, Wingfield BD (2013) Concerted evolution in the ribosomal RNA cistron. PLoS ONE 8:e59355CrossRefPubMedCentralPubMedGoogle Scholar
  30. Nkuekam KG, Wingfield MJ, Mohammed C, Carnegie AJ, Pegg GS, Roux J (2012) Ceratocystis species, including two new species associated with nitidulid beetles, on eucalypts in Australia. Anton Leeuw 101:217–241CrossRefGoogle Scholar
  31. Posada D, Crandall KA (1998) MODELTEST: testing the model of DNA substitution. Bioinformatics 14:817–818CrossRefPubMedGoogle Scholar
  32. Rayner RW (1970) A mycological colour chart. Commonwealth Mycological Institute, Surrey and British Mycological Society, KewGoogle Scholar
  33. Rodas CA, Roux J, Van Wyk M, Wingfield BD, Wingfield MJ (2008) Ceratocystis neglecta sp. nov. infecting Eucalyptus trees in Colombia. Fungal Divers 28:73–84Google Scholar
  34. Roux J, Wingfield MJ (2009) Ceratocystis species: emerging pathogens of non-native plantation Eucalyptus and Acacia species. South For 71:115–120Google Scholar
  35. Roux J, Wingfield MJ, Bouillet JP, Wingfield BD, Alfenas AC (1999) A serious new wilt disease of Eucalyptus caused by Ceratocystis fimbriata in Central Africa. For Pathol 30:175–184Google Scholar
  36. Roux J, Wingfield MJ, Bouillet JP, Winfield BD, Alfenas AC (2000) A serious new wilt disease of Eucalyptus caused by Ceratocystis fimbriata in Central Africa. For Pathol 30:175–184Google Scholar
  37. Roux J, Harrington TC, Steimel JP, Wingfield MJ (2001) Genetic variation in the wattle wilt pathogen Ceratocystis albifundus. Mycoscience 42:327–332CrossRefGoogle Scholar
  38. Roux J, Van Wyk M, Hatting H, Wingfield MJ (2004) Ceratocystis species infecting stem wounds on Eucalyptus grandis in South Africa. Plant Pathol 53:414–421CrossRefGoogle Scholar
  39. Roux J, Meke G, Kanyi B, Mwangi L, Mbaga A, Hunter GC, Nakabonge G, Heath RN, Wingfield MJ (2005) Diseases of plantation forestry trees species in Eastern Africa. S Afr J Sci 101:409–413Google Scholar
  40. Schoch CL, Seifert KA, Huhndorf S et al (2012) Nuclear ribosomal internal transcribed spacer (ITS) region as a universal DNA barcode marker for Fungi. Proc Natl Acad Sci USA 109:6241–6246CrossRefPubMedCentralPubMedGoogle Scholar
  41. Swofford DL (2002) PAUP* 4.0: phylogenetic analysis using parsimony (*and other methods). Sinauer Associates, SunderlandGoogle Scholar
  42. Sy CM (1956) Studies on the control of black rot (Ophiostoma fimbriatum) of sweet potato. Acta Phytopathol Sin 2:81–95 [In Chinese]Google Scholar
  43. Tamura K, Dudley J, Nei M, Kumar S (2007) MEGA 4: molecular evolutionary genetics analysis (MEGA) software version 4.0. Mol Biol Evol 24:1596–1599CrossRefPubMedGoogle Scholar
  44. Tsopelas P, Angelopoulos A (2004) First report of canker stain of plane strees, caused by Ceratocystis fimbriata sp. platani in Greece. Plant Pathol 53:531CrossRefGoogle Scholar
  45. Upadhyay HP (1981) A monograph of Ceratocystis and Ceratocystiopsis. University of Georgia Press, AthensGoogle Scholar
  46. Van Wyk M, Roux J, Barnes I, Wingfield BD, Liew ECY, Assa B, Summerell BA et al (2004) Ceratocystis polychroma sp. nov., a new species from Syzygium aromaticum in Sulawesi. Stud Mycol 50:273–282Google Scholar
  47. Van Wyk M, Al Adawi AO, Khan IA, Deadman ML, Al Jahwari AA, Wingfield BD, Ploetz R, Wingfield MJ (2007a) Ceratocystis manginecans sp. nov., causal agent of a destructive mango wilt disease in Oman and Pakistan. Fungal Divers 27:213–230Google Scholar
  48. Van Wyk M, Pegg G, Lawson S, Wingfield MJ (2007b) Ceratocystis atrox sp. nov. associated with Phoracantha acanthocera infestations on Eucalyptus grandis in Australia. Australas Plant Path 36:407–414CrossRefGoogle Scholar
  49. Van Wyk M, Wingfield BD, Clegg PA, Wingfield MJ (2009) Ceratocystis larium sp. nov., a new species from Styrax benzoin wounds associated with incense harvesting in Indonesia. Persoonia 22:75–82CrossRefPubMedCentralPubMedGoogle Scholar
  50. Van Wyk M, Roux J, Kamgan NG, Wingfield BD, Wingfield MJ (2012) Ceratocystis eucalypticola sp. nov. from Eucalyptus in South Africa and comparison to global isolates from this tree. IMA Fungus 3:45–58CrossRefPubMedCentralPubMedGoogle Scholar
  51. White TJ, Bruns T, Lee S, Taylor J (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis MA, Gelfand DH, Sninsky JJ, White TJ (eds) PCR protocols: a guide to methods and applications. Academic, New York, pp 230–257Google Scholar
  52. Wilken PM, Steenkamp ET, Wingfield MJ, De Beer ZW, Wingfield BD (2013) Draft nuclear genome sequence for the plant pathogen, Ceratocystis fimbriata. IMA Fungus 4:357–358CrossRefPubMedCentralPubMedGoogle Scholar
  53. Wingfield MJ, Seifert KA, Webber JF (1993) Ceratocystis and Ophiostoma: taxonomy, ecology and pathogenicity. American Phytopathological society Press, St PaulGoogle Scholar
  54. Wingfield MJ, De Beer ZW, Slippers B, Wingfield BD, Groenewald JZ, Lombard L, Crous PW (2012) One fungus, one name promotes progressive plant pathology. Mol Plant Pathol 13:604–613CrossRefPubMedGoogle Scholar
  55. Xie YJ (2011) Research progress on Eucalyptus breeding and its strategy in China. World Fore Res 24:50–54 [In Chinese]Google Scholar
  56. Xu B, Zheng XH, Guo WX, Zhou XP, He P (2011) First report of pomegranate wilt caused by Ceratocystis fimbriata in Sichuan Province. Plant Dis 95:776CrossRefGoogle Scholar
  57. Zhou XD, Wingfield MJ (2011) Eucalypt diseases and their management in China. Australas Plant Path 40:339–345CrossRefGoogle Scholar
  58. Zipfel RD, De Beer ZW, Jacobs K, Wingfield BD, Wingfield MJ (2006) Multigene phylogenies define Ceratocystis and Grosmannia distinct from Ophiostoma. Stud Mycol 55:75–79CrossRefPubMedCentralPubMedGoogle Scholar

Copyright information

© Springer International Publishing Switzerland 2015

Authors and Affiliations

  1. 1.Department of Microbiology and Plant Pathology, Forestry and Agricultural Biotechnology Institute (FABI)University of PretoriaPretoriaSouth Africa
  2. 2.China Eucalypt Research Centre (CERC)Chinese Academy of Forestry (CAF)ZhanJiangChina
  3. 3.Department of Genetics, Forestry and Agricultural Research Institute (FABI)University of PretoriaPretoriaSouth Africa

Personalised recommendations