European Journal of Plant Pathology

, Volume 152, Issue 3, pp 815–821 | Cite as

In vitro production of conidia of Elsinoë ampelina, the causal fungus of grapevine anthracnose

  • Ricardo Feliciano dos SantosEmail author
  • Marcel Bellato Spósito
  • Matthew Ayres
  • Mark Sosnowski


Anthracnose, caused by Elsinoë ampelina, is an important disease of grapevines in humid and warm production regions of the world. Colonies of E. ampelina grow slowly and rarely produce conidia on artificial media. To facilitate studies involving E. ampelina, our objective was to develop a method to induce significant conidial production of this fungus. In the present study, we induced the in vitro production of conidia of 10 Australian isolates by shake-incubation of mycelial fragments in rainwater under continuous light and darkness. Furthermore, seven Brazilian isolates were shake-incubated in rainwater and distilled water under continuous darkness. In both experiments cultures were shaken at 200-rpm and kept at room temperature (22–25 °C) for 7 days. Conidial production, germination, and severity of symptoms were quantified. Australian and Brazilian isolates produced different amounts of conidia. All Australian isolates sporulated in rainwater ranging from 4.20 × 103 to 5.91 × 106 conidia mL−1, with more than 90% conidial germination on water agar medium. All but one Brazilian isolate produced conidia in rainwater and distilled water, ranging from 7.50 × 103 to 8.22 × 106 conidia mL−1, with high germination percentages. Conidial suspensions from both Australian and Brazilian E. ampelina isolates caused typical anthracnose symptoms on grapevine leaves. This study describes an efficient method, using rainwater or distilled water associated with shaking, to induce the conidia production of E. ampelina.


Black spot Sporulation Rainwater Shaking Sphaceloma ampelinum Vitis vinifera 



This research was financially supported by São Paulo Research Foundation – FAPESP (grant numbers 2013/24003-9, 2014/24472-1 and 2016/01508-6). We thank Andrew Taylor (Department of Agriculture and Food, Western Australia), Bob Emmett (RWE Horticultural Pathology Research, Victoria), David Oag and James Drinnan (Department of Agriculture and Fisheries, Queensland) for providing anthracnose samples. We wish to acknowledge the South Australian Research and Development Institute (SARDI) for providing resources and staff for technical assistance. Scholarship provided by FAPESP (grant number 2016/01508-6) for the first author during his eight months in Australia is gratefully acknowledged.

Compliance with ethical standards

Conflict of interest

The authors declare no conflict of interest.

Supplementary material

10658_2018_1502_MOESM1_ESM.docx (42 kb)
Table S1 (DOCX 41 kb)


  1. Bagsic, I., Linde, M., & Debener, T. (2016). Genetic diversity and pathogenicity of Sphaceloma rosarum (teleomorph Elsinoë rosarum) causing spot anthracnose on roses. Plant Pathology, 65, 978–986.CrossRefGoogle Scholar
  2. Carisse, O., & Lefebvre, A. (2011). A model to estimate the amount of primary inoculum of Elsinoë ampelina. Plant Disease, 95, 1167–1171.CrossRefGoogle Scholar
  3. Elson, M. K., Schisler, D. A., & Jackson, M. A. (1998). Carbon-to-nitrogen ratio, carbon concentration, and amino acid composition of growth media influence conidiation of Helminthosporium solani. Mycologia, 90, 406–413.CrossRefGoogle Scholar
  4. EPPO (European and Mediterranean Plant Protection Organization) (2016). Elsinoe ampelina. Accessed 7 Dec 2017.
  5. Hyun, J., Paudyal, D. P., & Hwang, R. (2015). Improved method to increase conidia production from isolates of different pathotypes of citrus scab pathogen Elsinoe spp. Research in Plant Disease, 21, 231–234.CrossRefGoogle Scholar
  6. Kono, A., Nakaune, R., Yamada, M., Nakano, M., Mitani, N., & Ueno, T. (2009). Effect of culture conditions on conidia formation by Elsinoë ampelina, the causal organism of grapevine anthracnose. Plant Disease, 93, 481–484.CrossRefGoogle Scholar
  7. Magarey, R. D., Emmett, R. W., Magarey, P. A., & Franz, P. R. (1993). Evaluation of control of grapevine anthracnose caused by Elsinoe ampelina by pre-infection fungicides. Australasian Plant Pathology, 22, 48–52.CrossRefGoogle Scholar
  8. Mortensen, J. A. (1981). Sources and inheritance of resistance to anthracnose in Vitis. The Journal of Heredity, 72, 423–426.CrossRefGoogle Scholar
  9. R Core Team. (2016). R: A Language and environment for statistical computing. Vienna: R Foundation for Statistical Computing. Scholar
  10. Roncal, T., Cordobés, S., Sterner, O., & Ugalde, U. (2002). Conidiation in Penicillium cyclopium is induced by conidiogenone, an endogenous diterpene. Eukaryotic Cell, 1, 823–829.CrossRefGoogle Scholar
  11. Santos, R. F. (2017). Aetiology and epidemiology of grapevine anthracnose. Thesis: University of São Paulo.Google Scholar
  12. Santos, R. F., Ciampi-Guillardi, M., Amorim, L., Massola, N. S., & Spósito, M. B. (2018a). Aetiology of anthracnose on grapevine shoots in Brazil. Plant Pathology, 67, 692–706.CrossRefGoogle Scholar
  13. Santos, R. F., Spósito, M. B., Ayres, M. R., & Sosnowski, M. R. (2018b). Phylogeny, morphology and pathogenicity of Elsinoë ampelina, the causal agent of grapevine anthracnose in Brazil and Australia. Journal of Phytopathology, 166, 187–198.CrossRefGoogle Scholar
  14. Shear, C. L. (1929). The life history of Sphaceloma ampelinum de Bary. Phytopathology, 19, 673–679.Google Scholar
  15. Sosnowski, M., Emmett, B., Clarke, K., & Wicks, T. (2007). Susceptibility of tablegrapes to black spot (anthracnose) disease. The Australian & New Zealand Grapegrower & Winemaker, 521, 8–11.Google Scholar
  16. Sosnowski, M. R., Emmett, R. W., Wilcox, W. F., & Wicks, T. J. (2012). Eradication of black rot (Guignardia bidwellii) from grapevines by drastic pruning. Plant Pathology, 61, 1093–1102.CrossRefGoogle Scholar
  17. Su, Y., Qi, Y., & Cai, L. (2012). Induction of sporulation in plant pathogenic fungi. Mycology, 3, 195–200.Google Scholar
  18. Williamson, B., Hof, L., & Mcnicol, R. J. (1989). A method for in vitro production of conidia of Elsinoe veneta and the inoculation of raspberry cultivars. Annals of Applied Biology, 114, 23–33.CrossRefGoogle Scholar

Copyright information

© Koninklijke Nederlandse Planteziektenkundige Vereniging 2018

Authors and Affiliations

  • Ricardo Feliciano dos Santos
    • 1
    Email author
  • Marcel Bellato Spósito
    • 1
  • Matthew Ayres
    • 2
  • Mark Sosnowski
    • 2
    • 3
  1. 1.Departamento de Fitopatologia e Nematologia, Escola Superior de Agricultura “Luiz de Queiroz”Universidade de São PauloPiracicabaBrazil
  2. 2.South Australian Research and Development InstituteAdelaideAustralia
  3. 3.School of Agriculture, Food and Wine, Waite Research InstituteThe University of AdelaideAdelaideAustralia

Personalised recommendations