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European Journal of Plant Pathology

, Volume 124, Issue 4, pp 577–583 | Cite as

Cercospora zeina is the causal agent of grey leaf spot disease of maize in southern Africa

  • Barbara Meisel
  • Jeanne Korsman
  • Frederik J. Kloppers
  • Dave K. Berger
Article

Abstract

The aim of our study was to identify the causal agent of grey leaf spot disease of maize in southern Africa. Single-conidial cultures were recovered from maize leaves with typical disease symptoms sampled from several fields in South Africa, Zambia and Zimbabwe. Morphology, cultural characteristics, and a PCR-based test using Cercospora zeae-maydis and C. zeina-specific primer sets identified all single-conidial cultures as C. zeina. In addition, sequence alignment of DNA fragments of the internal transcribed spacer region (ITS1, ITS2, and the 5.8S gene) and elongation factor 1-α grouped all cultures in the same clade as the C. zeina ex-type culture CBS 118820. To by-pass cultivation of the slow-growing fungus, a rapid method to isolate DNA directly from lesions was successfully applied for PCR identification of C. zeina with species-specific ITS and histone primers. Koch’s postulates were fulfilled for C. zeina by artificially inoculating maize plants in a greenhouse, re-isolating conidia emerging from lesions and verifying pathogen identity with molecular techniques. These results provide evidence that confirms the presence of C. zeina and absence of C. zeae-maydis in commercial maize plantations in southern Africa.

Keywords

Diagnostic species-specific PCR assay Grey leaf spot Maize Koch’s postulates Cercospora zeina Cercospora zeae-maydis 

Abbreviations

CI

consistency index

EF

elongation factor 1-α

GLS

grey leaf spot

ITS

internal transcribed spacer

RI

retention index

Notes

Acknowledgements

We thank Martin Coetzee for assistance with the phylogenetic analysis. The funding of this work was provided by the Department of Science and Technology through the PlantBio Trust, South Africa and a Post-doctoral Fellowship from the University of Pretoria to BM.

References

  1. Altschul, S. F., Gish, W., Miller, W., Myers, E. W., & Lipman, D. J. (1990). Basic alignment search tool. Journal of Molecular Biology, 215, 403–410.PubMedGoogle Scholar
  2. Asea, G., Lipps, P. E., Pratt, R. C., Gordon, S. G., & Adipala, E. (2005). Development of greenhouse inoculation procedures for evaluation of partial resistance to Cercospora zeae-maydis in maize inbreds. Journal of Phytopathology, 153, 647–653. doi: 10.1111/j.1439-0434.2005.01032.x.CrossRefGoogle Scholar
  3. Bair, W., & Ayers, J. E. (1986). Variability in isolates of Cercospora zeae-maydis. Phytopathology, 76, 129–132. doi: 10.1094/Phyto-76-129.CrossRefGoogle Scholar
  4. Beckman, P. M., & Payne, G. A. (1982). External growth, penetration, and development of Cercospora zeae-maydis in corn leaves. Phytopathology, 72, 810–815. doi: 10.1094/Phyto-72-810.CrossRefGoogle Scholar
  5. Beckman, P. M., & Payne, G. A. (1983). Cultural techniques and conditions influencing growth and sporulation of Cercospora zeae-maydis and lesion development in corn. Phytopathology, 73, 286–289. doi: 10.1094/Phyto-73-286.CrossRefGoogle Scholar
  6. Carbone, I., & Kohn, L. M. (1999). A method for designing primer sets for speciation studies in filamentous ascomycetes. Mycologia, 91, 553–556. doi: 10.2307/3761358.CrossRefGoogle Scholar
  7. Chakrabarti, R., & Schutt, C. E. (2001). The enhancement of PCR amplification by low molecular weight amides. Nucleic Acids Research, 29, 2377–2381. doi: 10.1093/nar/29.11.2377.PubMedCrossRefGoogle Scholar
  8. Crous, P. W., Groenewald, J. Z., Groenewald, M., Caldwell, P., Braun, U., & Harrington, T. C. (2006). Species of Cercospora associated with grey leaf spot of maize. Studies in Mycology, 55, 189–197.PubMedCrossRefGoogle Scholar
  9. Dunkle, L. D., & Levy, M. (2000). Genetic relatedness of African and United States populations of Cercospora zeae-maydis. Phytopathology, 90, 486–490. doi: 10.1094/PHYTO.2000.90.5.486.PubMedCrossRefGoogle Scholar
  10. Goodwin, S. B., Dunkle, D. L., & Zismann, V. L. (2001). Phylogenetic analysis of Cercospora and Mycosphaerella based on the internal transcribed spacer region of ribosomal DNA. Phytopathology, 91, 648–658. doi: 10.1094/PHYTO.2001.91.7.648.PubMedCrossRefGoogle Scholar
  11. Griffith, G. W., & Shaw, D. S. (1998). Polymorphisms in Phytophthora infestans: Four Mitochondrial haplotypes are detected after PCR amplification of DNA from pure cultures or from host lesions. Applied and Environmental Microbiology, 64, 4007–4014.PubMedGoogle Scholar
  12. Latterell, F. M., & Rossi, A. E. (1983). Gray leaf spot of corn: A disease on the move. Plant Disease, 67, 842–847. doi: 10.1094/PD-67-842.CrossRefGoogle Scholar
  13. Möller, E. M., Bahnweg, G., Sandermann, H., & Geiger, H. H. (1992). A simple and efficient protocol for isolation of high molecular weight DNA from filament fungi. Nucleic Acids Research, 20, 6115–6116. doi: 10.1093/nar/20.22.6115.PubMedCrossRefGoogle Scholar
  14. Nowell, D. C. (1997) Studies on ear rot and gray leaf spot of maize in South Africa. Dissertation, University of NatalGoogle Scholar
  15. Okori, P., Fahleson, J., Rubaihayo, P. R., Adipala, E., & Dixelius, C. (2003). Assessment of genetic variation among East African Cercospora zeae-maydis. African Crop Science Journal, 11, 75–85.Google Scholar
  16. Paul, P. A., & Munkvold, G. P. (2005). Influence of temperature and relative humidity on sporulation of Cercospora zeae-maydis and expansion of gray leaf spot lesions on maize leaves. Plant Disease, 89, 624–630. doi: 10.1094/PD-89-0624.CrossRefGoogle Scholar
  17. Shim, W.-B., & Dunkle, D. L. (2005). Malazy, a degenerate, species-specific transposable element in Cercospora zeae-maydis. Mycologia, 97, 349–355. doi: 10.3852/mycologia.97.2.349.PubMedCrossRefGoogle Scholar
  18. Swofford, D. L. (2002). PAUP*: Phylogenetic analysis using parsimony (*and other methods). Version 4.1b10. Sunderland, MA: Sinauer.Google Scholar
  19. Tehon, L. R., & Daniels, E. (1925). Notes on parasitic fungi of Illinois. Mycologia, 17, 240–249. doi: 10.2307/3753890.CrossRefGoogle Scholar
  20. Wang, J., Levy, M., & Dunkle, L. D. (1998). Sibling species of Cercospora associated with gray leaf spot of maize. Phytopathology, 88, 1269–1275. doi: 10.1094/PHYTO.1998.88.12.1269.PubMedCrossRefGoogle Scholar
  21. Ward, J. M. J. (1996) Epidemiology and management of gray leaf spot: A new disease of maize in South Africa. Dissertation, University of NatalGoogle Scholar
  22. Ward, J. M. J., Laing, M. D., & Cairns, A. L. P. (1997). Management practices to reduce gray leaf spot of maize. Crop Science, 37, 1257–1262.CrossRefGoogle Scholar
  23. Ward, J. M. J., Stromberg, E. L., Nowell, D. C., & Nutter Jr., F. W. (1999). Gray leaf spot. A disease of global importance in maize production. Plant Disease, 83, 884–895. doi: 10.1094/PDIS.1999.83.10.884.CrossRefGoogle Scholar
  24. White, T. J., Bruns, T., Lee, S., & Taylor, J. (1990). Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In M. A. Innis, D. H. Gelfand, J. J. Sninsky, & T. J. White (Eds.),PCR protocols: S guide to methods and applications (pp. 315–322). California: Academic.Google Scholar

Copyright information

© KNPV 2009

Authors and Affiliations

  • Barbara Meisel
    • 1
  • Jeanne Korsman
    • 1
  • Frederik J. Kloppers
    • 2
  • Dave K. Berger
    • 1
  1. 1.Department of Plant Science, Forestry and Agricultural Biotechnology Institute (FABI)University of PretoriaPretoriaSouth Africa
  2. 2.PANNAR Research Services (Pty) LtdGreytownSouth Africa

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