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Concomitant reiterative BAC walking and fine genetic mapping enable physical map development for the broad-spectrum late blight resistance region, RB

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Abstract

The wild potato species Solanum bulbocastanum is a source of genes for potent late blight resistance. We previously mapped resistance to a single region of the S. bulbocastanum chromosome 8 and named the region RB (for "Resistance from S. B ulbocastanum "). We now report physical mapping and contig construction for the RB region via a novel reiterative method of BAC walking and concomitant fine genetic mapping. BAC walking was initiated using RFLP markers previously shown to be associated with late blight resistance. Subcontig extension was accomplished using new probes developed from BAC ends. Significantly, BAC end and partial BAC sequences were also used to develop PCR-based markers to enhance map resolution in the RB region. As they were developed from BAC clones of known position relative to RB, our PCR-based markers are known a priori to be physically closer to the resistance region. These markers allowed the efficient screening of large numbers of segregating progeny at the cotyledon stage, and permitted us to assign the resistance phenotype to a region of approximately 55 kb. Our markers also directed BAC walking efforts away from regions distantly related to RB in favor of the 55-kb region. Because the S. bulbocastanum genotype used in BAC library construction is heterozygous for RB (RB/rb), codominant PCR-based markers, originally developed for fine-scale mapping, were also used to determine homolog origins for individual BAC clones. Ultimately, BAC contigs were constructed for the RB region from both resistant (RB) and susceptible (rb) homologs.

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Acknowledgements

This work was funded in part by NSF Plant Genome Project #DBI-9975866. Mention of a trademark, proprietary product, or vendor does not constitute a guarantee or warranty of the product by the USDA, and does not imply its approval to the exclusion of other products and vendors that might also be suitable

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Author notes

  1. J. M. Bradeen

    Present address: Department of Plant Pathology, University of Minnesota, 495 Borlaug Hall, 1991 Upper Buford Circle, St. Paul, MN 55108, USA

  2. S. K. Naess

    Present address: Centre de Recherche Les Buissons, 358 rue Principale, Pointe-aux-Outardes, Québec, G0H 1M0, Canada

Authors and Affiliations

  1. Department of Plant Pathology, University of Wisconsin, 1630 Linden Drive, Madison, WI 53706, USA

    J. M. Bradeen & S. M. Wielgus

  2. USDA-ARS, Department of Plant Pathology, University of Wisconsin, 1630 Linden Drive, Madison, WI 53706, USA

    S. K. Naess, G. T. Haberlach & J. P. Helgeson

  3. The Institute for Genomic Research, 9712 Medical Center Drive, Rockville, MD 20850, USA

    C. R. Buell

  4. Department of Horticulture, University of Wisconsin, 1575 Linden Drive, Madison, WI 53706, USA

    J. Song & J. Jiang

Authors
  1. J. M. Bradeen
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  2. S. K. Naess
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  3. J. Song
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  4. G. T. Haberlach
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  5. S. M. Wielgus
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  6. C. R. Buell
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  7. J. Jiang
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  8. J. P. Helgeson
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Corresponding authors

Correspondence to J. Jiang or J. P. Helgeson.

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Communicated by R. Hagemann

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Bradeen, J.M., Naess, S.K., Song, J. et al. Concomitant reiterative BAC walking and fine genetic mapping enable physical map development for the broad-spectrum late blight resistance region, RB . Mol Gen Genomics 269, 603–611 (2003). https://doi.org/10.1007/s00438-003-0865-8

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  • DOI: https://doi.org/10.1007/s00438-003-0865-8

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