Microsatellite and mating type primers for the maize and sorghum pathogen, Exserohilum turcicum
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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.
KeywordsFungi 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.
- Kloppers R, Tweer S (2009) Northern corn leaf blight fact sheet. PANNAR Seed (Pty) LtdGoogle Scholar
- 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
- 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
- Simcox KD, Pedersen WL, Nickrent DL (1993) Isozyme diversity in Setosphaeria turcica. Can J Plant Pathol 15:91–96Google Scholar
- 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