Australasian Plant Pathology

, Volume 43, Issue 5, pp 577–581 | Cite as

Microsatellite and mating type primers for the maize and sorghum pathogen, Exserohilum turcicum

  • M. P. Haasbroek
  • M. Craven
  • I. Barnes
  • B. G. CramptonEmail author


Northern corn leaf blight (NCLB) is a destructive foliar disease of maize that results from infection with the fungal pathogen, Exserohilum turcicum. Annual yield losses incurred from NCLB in South Africa may exceed 50 % when environmental conditions optimal for disease development prevail. In order to study the genetic diversity of E. turcicum, 13 microsatellite markers and mating type PCR primers were developed. Thirty-two primer pairs were designed from the E. turcicum genome sequence to flank microsatellite regions. A multiplex PCR assay amplifying both mating type idiomorphs was designed from the MAT1-1 and MAT1-2 gene sequences, and a protocol for multiplex PCR amplification of MAT loci was optimized. Initial screening identified 13 microsatellite regions that were polymorphic in 9 isolates of E. turcicum. To test the efficacy of the markers, 26 isolates of E. turcicum from 6 South African provinces, including 2 isolates from sorghum, were genotyped. A total of 90 alleles across 13 loci were obtained and the gene diversity ranged from 0.074 to 0.929. Cross-species amplification with E. rostratum was obtained for one SSR marker (SSR27). The MAT markers were specific to E. turcicum and could be used to differentiate isolates of E. turcicum and E. rostratum. The markers developed in this study will be useful to elucidate the population genetic structure, genetic diversity and mode of reproduction of E. turcicum on maize and sorghum.


Fungi Mating type markers Microsatellite Multiplex PCR Northern corn leaf blight (NCLB) Setosphaeria turcica 



This work is based on the research supported in part by the National Research Foundation of South Africa (grant specific unique reference number (UID) 85076), and by the Maize Trust and the University of Pretoria’s Research and Development Programme. The financial assistance of the National Research Foundation (NRF) towards this research is hereby acknowledged. Opinions expressed and conclusions arrived at, are those of the author and are not necessarily to be attributed to the NRF. The authors are grateful to the Genomics Research Institute for financial support to Maria Haasbroek. The authors would like to thank Ms Renate Zipfel and Ms Gladys Shabangu of the sequencing facility at the University of Pretoria for technical assistance.

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  1. Agapow PM, Burt A (2001) Indices of multilocus linkage disequilibrium. Mol Ecol Notes 1:101–102CrossRefGoogle Scholar
  2. Bentolila S, Guitton C, Bouvet N, Sailland A, Nykaza S, Freyssinet G (1991) Identification of an RFLP marker tightly linked to the Ht1 gene in maize. Theor Appl Genet 82:393–398PubMedCrossRefGoogle Scholar
  3. Carson ML (1995) Inheritance of latent period length in maize with Exserohilum turcicum. Plant Dis 79:581–585CrossRefGoogle Scholar
  4. Chambers GK, MacAvoy ES (2000) Microsatellites: consensus and controversy. Comp Biochem Physiol 126:455–476CrossRefGoogle Scholar
  5. Craven M, Fourie AP (2011) Field evaluation of maize inbred lines for resistance to Exserohilum turcicum. S Afr J Plant Soil 28:69–74CrossRefGoogle Scholar
  6. Dong J, Fan Y et al (2008) Geographic distribution and genetic analysis of physiological races of Setosphaeria turcica in Northern China. Am J Agric Biol Sci 3:389–398CrossRefGoogle Scholar
  7. Ellegren H (2004) Microsatellites: simple sequences with complex evolution. Nat Rev Genet 5:435–445PubMedCrossRefGoogle Scholar
  8. Faircloth BC (2008) MSATCOMMANDER: detection of microsatellite repeat arrays and automated, locus-specific primer design. Mol Ecol Resour 8:92–94PubMedCrossRefGoogle Scholar
  9. Ferguson LM, Carson ML (2004) Spatial diversity of Setosphaeria turcica sampled from the eastern United States. Phytopathology 94:892–900PubMedCrossRefGoogle Scholar
  10. Ferguson LM, Carson ML (2007) Temporal variation in Setosphaeria turcica between 1974 and 1994 and origin of races 1, 23, and 23N in the United States. Phytopathology 97:1501–1511PubMedCrossRefGoogle Scholar
  11. Kerenyi Z, Zeller K, Hornok L, Leslie JF (1999) Molecular standardization of mating type terminology in the Gibberella fujikuroi species complex. Appl Environ Microbiol 65:4071–4076PubMedCentralPubMedGoogle Scholar
  12. Kloppers R, Tweer S (2009) Northern corn leaf blight fact sheet. PANNAR Seed (Pty) LtdGoogle Scholar
  13. Leonard KJ, Suggs EG (1974) Setosphaeria prolata, ascigerous state of Exserohilum prolatum. Mycologia 66:281–297CrossRefGoogle Scholar
  14. Levy Y (1991) Variation in fitness among field isolates of Exserohilum turcicum in Israel. Plant Dis 75:163–166CrossRefGoogle Scholar
  15. McDonald BA, McDermott JM (1993) Population genetics of plant pathogenic fungi. Bioscience 43:311–319CrossRefGoogle Scholar
  16. Muiru WM, Koopmann B, Tiedemann AV, Mutitu EW, Kimenju JW (2010) Evaluation of genetic variability of Kenyan, German and Austrian isolates of Exserohilum turcicum using amplified fragment length polymorphism DNA markers. Biotechnology 9:204–211Google Scholar
  17. Murray MG, Thompson WF (1980) Rapid isolation of high molecular weight plant DNA. Nucleic Acids Res 8:4321–4325PubMedCentralPubMedCrossRefGoogle Scholar
  18. Nei M (1973) Analysis of gene diversity in subdivided populations. Proc Natl Acad Sci U S A 70:3321–3323PubMedCentralPubMedCrossRefGoogle Scholar
  19. Ohm RA, Feau N et al (2012) Diverse lifestyles and strategies of plant pathogenesis encoded in the genomes of eighteen Dothideomycetes fungi. PLoS Pathog 8:1–26CrossRefGoogle Scholar
  20. Ramathani I, Biruma M, Martin T, Dixelius C, Okori P (2011) Disease severity, incidence and races of Setosphaeria turcica on sorghum in Uganda. Eur J Plant Pathol 131:383–392CrossRefGoogle Scholar
  21. Rozen S, Skaletsky H (2000) Primer3 on the WWW for general users and for biologist programmers. Methods in molecular biology (Clifton, N.J.) pp 365–86Google Scholar
  22. Simcox KD, Pedersen WL, Nickrent DL (1993) Isozyme diversity in Setosphaeria turcica. Can J Plant Pathol 15:91–96Google Scholar
  23. White TJ, Bruns TD, Lee SB, Taylor JW (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: PCR protocols: a guide to methods and applications. Academic, New York, pp 315–322CrossRefGoogle Scholar
  24. Yeh FC, Yang RC, Boyle T, Ye ZH, Mao JX (1997) POPGENE: The user-friendly shareware for population genetic analysis. University of Alberta, Canada: Molecular Biology and Biotechnology CentreGoogle Scholar

Copyright information

© Australasian Plant Pathology Society Inc. 2014

Authors and Affiliations

  • M. P. Haasbroek
    • 1
  • M. Craven
    • 3
  • I. Barnes
    • 2
  • B. G. Crampton
    • 1
    Email author
  1. 1.Department of Plant Science, Forestry and Agricultural Biotechnology Institute (FABI)University of PretoriaPretoriaSouth Africa
  2. 2.Department of Genetics, Forestry and Agricultural Biotechnology Institute (FABI)University of PretoriaPretoriaSouth Africa
  3. 3.Agricultural Research Council-Grain Crops InstitutePotchefstroomSouth Africa

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