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Development and mapping of microsatellite (SSR) markers in wheat

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Abstract

Microsatellite DNA markers are consistently found to be more informative than other classes of markers in hexaploid wheat. The objectives of this research were to develop new primers flanking wheat microsatellites and to position the associated loci on the wheat genome map by genetic linkage mapping in the ITMI W7984 × Opata85 recombinant inbred line (RIL) population and/or by physical mapping with cytogenetic stocks. We observed that the efficiency of marker development could be increased in wheat by creating libraries from sheared rather than enzyme-digested DNA fragments for microsatellite screening, by focusing on microsatellites with the [ATT/TAA] n motif, and by adding an untemplated G-C clamp to the 5′-end of primers. A total of 540 microsatellite-flanking primer pairs were developed, tested, and annotated from random genomic libraries. Primer pairs and associated loci were assigned identifiers prefixed with BARC (the acronym for the USDA-ARS Beltsville Agricultural Research Center) or Xbarc, respectively. A subset of 315 primer sets was used to map 347 loci. One hundred and twenty-five loci were localized by physical mapping alone. Of the 222 loci mapped with the ITMI population, 126 were also physically mapped. Considering all mapped loci, 126, 125, and 96 mapped to the A, B, and D genomes, respectively. Twenty-three of the new loci were positioned in gaps larger than 10 cM in the map based on pre-existing markers, and 14 mapped to the ends of chromosomes. The length of the linkage map was extended by 80.7 cM. Map positions were consistent for 111 of the 126 loci positioned by both genetic and physical mapping. The majority of the 15 discrepancies between genetic and physical mapping involved chromosome group 5.

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Acknowledgements

The authors wish to thank Agricultural Technology, Utilization and Transfer (ATUT) Project 008, the Steering Committee of the U.S. Wheat and Barley Scab Initiative, and the USDA-ARS for the support of this research. Also, the excellent technical assistance of Susan Fogarty and P. Chris Lee is gratefully acknowledged. Victoria Carollo of GrainGenes provided invaluable assistance in identifying errors in the names of loci.

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  1. J. R. Shi

    Present address: Jiangsu Academy of Agricultural Sciences, Nanjing, 210014, P.R. China

Authors and Affiliations

  1. Soybean Genomics and Improvement Lab, Beltsville Agricultural Research Center, USDA-ARS, MD, 20705, USA

    Q. J. Song, E. W. Fickus & P. B. Cregan

  2. Department of Natural Resource Sciences & Landscape Architecture, University of Maryland, MD, 20742, USA

    Q. J. Song & J. M. Costa

  3. Department of Crop and Soil Science, Michigan State University, East Lansing, MI, 48824, USA

    J. R. Shi, J. Lewis & R. Ward

  4. Department of Plant Pathology, Kansas State University, Manhattan, KS, 66506, USA

    S. Singh & B. S. Gill

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  1. Q. J. Song
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Song, Q.J., Shi, J.R., Singh, S. et al. Development and mapping of microsatellite (SSR) markers in wheat. Theor Appl Genet 110, 550–560 (2005). https://doi.org/10.1007/s00122-004-1871-x

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