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

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Abstract

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.

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Acknowledgments

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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Authors and Affiliations

  1. Department of Plant Science, Forestry and Agricultural Biotechnology Institute (FABI), University of Pretoria, Pretoria, 0002, South Africa

    M. P. Haasbroek & B. G. Crampton

  2. Department of Genetics, Forestry and Agricultural Biotechnology Institute (FABI), University of Pretoria, Pretoria, 0002, South Africa

    I. Barnes

  3. Agricultural Research Council-Grain Crops Institute, Potchefstroom, 2520, South Africa

    M. Craven

Authors
  1. M. P. Haasbroek
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  2. M. Craven
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  3. I. Barnes
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  4. B. G. Crampton
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Corresponding author

Correspondence to B. G. Crampton.

Additional information

I. Barnes and B. G. Crampton are senior authors who contributed equally to this article.

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Haasbroek, M.P., Craven, M., Barnes, I. et al. Microsatellite and mating type primers for the maize and sorghum pathogen, Exserohilum turcicum . Australasian Plant Pathol. 43, 577–581 (2014). https://doi.org/10.1007/s13313-014-0289-4

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  • DOI: https://doi.org/10.1007/s13313-014-0289-4

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