Skip to main content
SpringerLink
Log in

Multi-gene sequence analysis and phenotypic diversity of Phaeoacremonium species isolated from grapevines in Spain

  • Published:
European Journal of Plant Pathology Aims and scope Submit manuscript

Abstract

Esca and Petri disease, caused by Phaeoa-cremonium, and other grapevine decline diseases, such as eutypa dieback, are responsible for serious economic losses to the wine industry worldwide. In the present work, 175 isolates of Phaeoacremonium spp. were collected (mostly in the region of Castilla y León, Spain) from vines either asymptomatic or symptomatic for grapevine decline diseases. The sequences of DNA fragments of 30 isolates were analysed. Amplicons of ITS, and partial gene of the β-tubulin, actin, calmodulin and translation elongation factor 1-α [EF1-α], were obtained by PCR. This is the first time that any exon of the EF1-α gene or exons 6–7 of the β-tubulin gene of seven Phaeoacremonium spp. have been amplified. Amplicons of the calmodulin and EF1-α exons were the most informative in terms of species differentiation: only 52.6 % and 56.6 % similarity across six and seven species was respectively seen. Phylogenetic analysis of combined DNA sequences revealed five Phaeoacremonium species isolated from the sampled plants; P. aleophilum was the most common. Four other species were found, however only one isolate each. P. mortoniae and P. iranianum are here reported for the first time on grapevines in the region of Castilla y León and the neighbouring region of La Rioja respectively. In total, 29 nucleotide differences among the studied gene fragments were seen across 23 isolates of P. aleophilum. Network analysis showed these Spanish P. aleophilum isolates to fall into two main groups of genotypes. P. aleophilum genotypes belonging to group 1 were mostly isolated from young grapevines affected by Petri disease but genotypes included in group 2 were isolated from adult plants showing esca or eutypa dieback symptoms.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  • Alaniz, S., León, M., Vicent, A., García-Jiménez, J., Abad-Campos, P., & Armengol, J. (2007). Characterization of Cylindrocarpon species associated with black foot disease of grapevine in Spain. Plant Disease, 91, 1187–1193.

    Article  CAS  Google Scholar 

  • Aroca, A., & Raposo, R. (2007). PCR-based strategy to detect and identify species of Phaeoacremonium causing grapevine diseases. Applied and Environmental Microbiology, 73, 2911–2918.

    Article  PubMed  CAS  Google Scholar 

  • Aroca, A., García-Figueres, F., Bracamonte, L., Luque, J., & Raposo, R. (2006). A survey of trunk disease pathogens within rootstocks of grapevines in Spain. European Journal of Plant Pathology, 115, 195–202.

    Article  Google Scholar 

  • Aroca, A., Garcia-Figueres, F., Bracamonte, L., Luque, J., & Raposo, R. (2007). First report of Phaeoacremonium viticola affecting grapevines in Spain. Plant Pathology, 57, 386.

    Article  Google Scholar 

  • Bandelt, H. J., Forster, P., Sykes, B. C., & Richards, M. B. (1995). Mitochondrial portraits of human populations using median networks. Genetics, 141, 743–753.

    PubMed  CAS  Google Scholar 

  • Bandelt, H. J., Forster, P., & Rohl, A. (1999). Median-joining networks for inferring intraspecific phylogenies. Molecular Biology and Evolution, 16, 37–48.

    Article  PubMed  CAS  Google Scholar 

  • Berraf-Tebbal, A., Bouznad, Z., Santos, J. M., Coelho, M. A., Péros, J. P., & Phillips, A. J. L. (2011). Phaeoacremonium species associated with Eutypa dieback and esca of grapevines in Algeria. Phytopathologia Mediterranea, 50(Supplement), S86–S97.

    Google Scholar 

  • Carbone, I., & Kohn, L. M. (1999). A method for designing primer sets for speciation studies in filamentous ascomycetes. Mycologia, 91, 553–556.

    Article  CAS  Google Scholar 

  • Carter, M.V. (1976). Biennal report of the Waite Agricultural research Institute, South Australia, for 1974–1975, pp. 96–7.

  • Cobos, R., & Martín, M. T. (2008). Molecular characterisation of Phaeomoniella chlamydospora isolated from grapevines in Castilla y León (Spain). Phytopathologia Mediterranea, 47, 20–27.

    CAS  Google Scholar 

  • Crous, P. W., Gams, W., Wingfield, M. J., & van Wyk, P. S. (1996). Phaeoacremonium gen. nov. associated with wilt and decline diseases of woody hosts and human infections. Mycologia, 88, 786–796.

    Article  Google Scholar 

  • Dupont, J., Laloui, W., & Magnin, S. (2000). Phaeoacremonium viticola, a new species associated with Esca disease of grapevine in France. Mycologia, 92, 499–504.

    Article  Google Scholar 

  • Dupont, J., Magnin, S., Césari, C., & Gatica, M. (2002). ITS and β-tubulin markers help delineate Phaeoacremonium species, and the occurrence of P. parasiticum in grapevine disease in Argentina. Mycological Research, 106, 1143–1150.

    Article  CAS  Google Scholar 

  • Edwards, J., & Pascoe, I. (2004). Occurrence of Phaeomoniella chlamydospora and Phaeoacremonium aleophilum associated with Petri disease and esca in Australian grapevines. Australasian Plant Pathology, 33, 273–279.

    Article  Google Scholar 

  • Essakhi, S., Mugnai, L., Crous, P. W., Groenewald, J. Z., & Surico, G. (2008). Molecular and phenotypic characterisation of novel Phaeoacremonium species isolated from esca diseased grapevines. Persoonia, 21, 119–134.

    Article  PubMed  CAS  Google Scholar 

  • Fischer, M., & Kassemeyer, H. H. (2003). Fungi associated with esca disease of grapevine in Germany. Vitis, 42, 109–116.

    CAS  Google Scholar 

  • Glass, N. L., & Donaldson, G. C. (1995). Development of primer sets designed for use with the PCR to amplify conserved genes from filamentous ascomycetes. Applied and Environmental Microbiology, 61, 1323–1330.

    PubMed  CAS  Google Scholar 

  • Graham, A. B., Johnston, P. R., & Weir, B. S. (2009). Three new Phaeoacremonium species on grapevines in New Zealand. Australasian Plant Pathology, 93, 505–513.

    Article  Google Scholar 

  • Gramaje, D., Alainz, S., Pérez-Sierra, A., Abad-Campos, P., García-Jiménez, J., & Armengol, J. (2007). First report of Phaeoacremonium mortoniae causing Petri disease of grapevine in Spain. Plant Disease, 91, 1206.

    Article  Google Scholar 

  • Gramaje, D., Armengol, J., Colino, M. J., Santiago, R., Moralejo, E., Luque, J., & Mostert, L. (2009a). First report of Phaeoacremonium inflatipes, P. iranianum, and P. sicilianum causing Petri disease of grapevine in Spain. Plant Disease, 93, 964.

    Article  Google Scholar 

  • Gramaje, D., Armengol, J., Mohammadi, H., Banihashemi, Z., & Mostert, L. (2009b). Novel Phaeoacremonium species associated with Petri disease and esca of grapevine in Iran and Spain. Mycologia, 101, 920–929.

    Article  PubMed  Google Scholar 

  • Groenewald, M., Kang, J. C., Crous, P. W., & Gams, W. (2001). ITS and β-tululin phylogeny of Phaeoacremonium and Phaeomoniella species. Mycological Research, 105, 651–657.

    Article  CAS  Google Scholar 

  • Hasegawa, M., Kishino, H., & Yano, T. (1985). Dating the human-ape splitting by a molecular clock of mitochondrial DNA. Journal of Molecular Evolution, 22, 160–174.

    Article  PubMed  CAS  Google Scholar 

  • Lardner, R., Stummer, B., Sosnowski, M., & Scott, E. (2005). Molecular identification and detection of Eutypa lata in grapevine. Mycological Research, 109, 799–808.

    Article  PubMed  CAS  Google Scholar 

  • Larignon, P., & Dubos, B. (1997). Fungi associated with esca disease in grapevine. European Journal of Plant Pathology, 103, 147–157.

    Article  Google Scholar 

  • Lecomte, P., Péros, J. P., Blancard, D., Bastien, N., & Délye, C. (2000). PCR assays that identify the grapevine dieback fungus Eutypa lata. Applied and Environmental Microbiology, 66, 4475–4480.

    Article  PubMed  CAS  Google Scholar 

  • Martín, M. T., & Cobos, R. (2007). Identification of fungi associated with grapevine decline in Castilla y León (Spain). Phytopathologia Mediterranea, 46, 18–25.

    Google Scholar 

  • Martín, L., Cuesta, M. J., & Martín, M. T. (2011). A new report of Phaeoacremonium viticola and P. hispanicum causing grapevine trunk disease in Castilla y León, Spain. New Disease Reports, 23, 27. doi:10.5197/j.2044-0588.2011.023.027.

    Article  Google Scholar 

  • Martín, M. T., Cobos, R., Martín, L., & López-Enríquez, L. (2012). Real-time PCR detection of Phaeomoniella chlamydospora and Phaeoacremonium aleophilum. Applied and Environmental Microbiology, 78, 3985–3991.

    Article  PubMed  Google Scholar 

  • Mostert, L., Crous, P. W., Groenewald, J. Z., Gams, W., & Summerbell, R. (2003). Togninia (Calosphaeriales) is confirmed as teleomorph of Phaeoacremonium by means of morphology, sexual compatibility, and DNA phylogeny. Mycologia, 95, 646–659.

    Article  PubMed  Google Scholar 

  • Mostert, L., Groenewald, J. Z., Summerbell, R. C., Robert, V., Sutton, D. A., Padhye, A. A., & Crous, P. W. (2005). Species of Phaeoacremonium associated with human infections and environmental reservoirs in infected woody plants. Journal of Clinical Microbiology, 43, 1752–1767.

    Article  PubMed  Google Scholar 

  • Mostert, L., Groenewald, J. Z., Summerbell, R. C., Gams, W., & Crous, P. W. (2006). Taxonomy and pathology of Togninia (Diaporthales) and its Phaeoacremonium anamorphs. Studies in Mycology, 54, 1–115.

    Article  Google Scholar 

  • Mugnai, L., Graniti, A., & Surico, G. (1999). Esca (black measles) and brown wood-streaking: two old and elusive diseases of grapevines. Plant Disease, 83, 404–418.

    Article  Google Scholar 

  • Muruamendiaraz, A., Lecomte, P., & Legorburu, F. J. (2009). Occurrence of Eutypa lata sexual stage on grapevine in Rioja. Phytopathologia Mediterranea, 48, 140–144.

    Google Scholar 

  • Nei, M., & Kumar, S. (2000). Molecular evolution and phylogenetics. New York: Oxford University Press.

    Google Scholar 

  • O’Donnell, K., & Cigelinik, E. (1997). Two different intragenomic rDNA ITS2 types within a monophyletic lineage of the fungus Fusarium are nonorthologous. Molecular Phylogenetics and Evolution, 7, 103–116.

    Article  PubMed  Google Scholar 

  • Pascoe, I., & Cottral, E. (2000). Developments in grapevine trunk diseases research in Australia. Phytopathologia Mediterranea, 39, 68–75.

    Google Scholar 

  • Petit, E., & Gubler, W. D. (2005). Characterization of Cylindrocarpon species, the cause of black foot disease of grapevine in California. Plant Disease, 89, 1051–1059.

    Article  CAS  Google Scholar 

  • Rooney-Latham, S., Eskalen, A., & Gubler, W. D. (2005). Teleomorph formation of Phaeoacremonium aleophilum, cause of esca and grapevine decline in California. Plant Disease, 89, 177–184.

    Article  Google Scholar 

  • Scheck, H. J., Vasquez, S. J., & Gubler, W. D. (1998). First report of three Phaeoacremonium spp., causing young grapevine decline in California. Plant Disease, 82, 590.

    Article  Google Scholar 

  • Surico, G., Mugnai, L., & Marchi, G. (2006). Older and more recent observations on esca: a critical overview. Phytopathologia Mediterranea, 45(Supplement), S68–S86.

    Google Scholar 

  • Swofford, D. L. (2000). PAUP* 4.0: Phylogenetic analysis using parsimony. Sunderland: Sinauer Associates.

    Google Scholar 

  • Tamura, K., Peterson, D., Peterson, N., Stecher, G., Nei, M., & Kumar, S. (2011). MEGA 5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Molecular Biology and Evolution, 28(10), 2731–2739. doi:10.1093/molbev/msr121.

    Article  PubMed  CAS  Google Scholar 

  • Tegli, S., Bertelli, E., & Surico, G. (2000). Sequence analysis of ITS ribosomal DNA in five Phaeoacremonium species and development of a PCR-based assay for the detection of P. chlamydosporum and P. aleophilum in grapevine tissue. Phytopathologia Mediterranea, 39, 134–149.

    CAS  Google Scholar 

  • White, T. J., Bruns, T., Innis, M. A., Lee, S., & Taylor, J. (1990). Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: PCR protocols: A guide to methods and applications, pp 315–322.

  • Whiting, E. C., Khan, A., & Gubler, W. D. (2001). Effect of temperature and water potential on survival and mycelial growth of Phaeomoniella chlamydospora and Phaeoacremonium spp. Plant Disease, 85, 195–201.

    Article  Google Scholar 

Download references

Acknowledgments

This work was funded by the Instituto Nacional de Investigación y Tecnología Agraria y Alimentarias (RTA04-127 and RTA2007-07) and by FEDER funds. Laura Martín was supported by the Programa Personal Técnico de Apoyo, Ministerio de Educación y Ciencia (grant PTA-2003-01-01001). Maria Teresa Martín was supported by the Contratos Doctores Sistema INIA-CCAA grants DR02-272 and DR07-0161. María José Cuesta and Miguel Ángel Sanz provided helpful insight.

Author information

Authors and Affiliations

  1. Instituto Tecnológico Agrario de Castilla y León (ITACYL), Ctra. de Burgos Km 119, 47071, Valladolid, Spain

    Laura Martín & María Teresa Martín

Authors
  1. Laura Martín
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. María Teresa Martín
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to María Teresa Martín.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Martín, L., Martín, M.T. Multi-gene sequence analysis and phenotypic diversity of Phaeoacremonium species isolated from grapevines in Spain. Eur J Plant Pathol 137, 343–361 (2013). https://doi.org/10.1007/s10658-013-0247-y

Download citation

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10658-013-0247-y

Keywords

Search

Navigation