Molecular Genetics and Genomics

, Volume 269, Issue 5, pp 603–611 | Cite as

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

  • J. M. Bradeen
  • S. K. Naess
  • J. Song
  • G. T. Haberlach
  • S. M. Wielgus
  • C. R. Buell
  • J. Jiang
  • J. P. Helgeson
Original Paper

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. Bulbocastanum "). 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.

Keywords

Solanum bulbocastanum General resistance Reiterative mapping Cleaved Amplified Polymorphic Sequences (CAPS) 

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Copyright information

© Springer-Verlag 2003

Authors and Affiliations

  • J. M. Bradeen
    • 1
    • 5
  • S. K. Naess
    • 2
    • 6
  • J. Song
    • 4
  • G. T. Haberlach
    • 2
  • S. M. Wielgus
    • 1
  • C. R. Buell
    • 3
  • J. Jiang
    • 4
  • J. P. Helgeson
    • 2
  1. 1.Department of Plant PathologyUniversity of WisconsinMadisonUSA
  2. 2.USDA-ARS, Department of Plant PathologyUniversity of WisconsinMadisonUSA
  3. 3.The Institute for Genomic ResearchRockvilleUSA
  4. 4.Department of HorticultureUniversity of WisconsinMadisonUSA
  5. 5.Department of Plant PathologyUniversity of MinnesotaSt. PaulUSA
  6. 6.Centre de Recherche Les BuissonsPointe-aux-OutardesCanada

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