Plant Molecular Biology

, Volume 54, Issue 5, pp 727–742 | Cite as

Chromosome Walking in the Petunia Inflata Self-Incompatibility (S-) Locus and Gene Identification in an 881-kb Contig Containing S2-RNase

  • Yan Wang
  • Tatsuya Tsukamoto
  • Ki-wan Yi
  • Xi Wang
  • Shihshieh Huang
  • Andrew G. McCubbin
  • Teh-hui Kao


Self-incompatibility (SI) in the Solanaceae, Rosaceae and Scrophulariaceae is controlled by the polymorphic S locus, which contains two separate genes encoding pollen and pistil determinants in SI interactions. The S-RNase gene encodes the pistil determinant, whereas the pollen determinant gene, named the pollen S gene, has not yet been identified. Here, we set out to construct an integrated genetic and physical map of the S locus of Petunia inflata and identify any additional genes located at this locus. We first conducted chromosome walking at the S2 locus using BAC clones that contained either S2-RNase or one of the nine markers tightly linked to the S locus. Ten separate contigs were constructed, which collectively spanned 4.4 Mb. To identify additional genes located at the S2 locus, a 328-kb region (part of an 881-kb BAC contig) containing S2-RNase was completely sequenced. Approximately 76% of the region contained repetitive sequences, including transposon-like sequences. Other than S2-RNase, an F-box gene, named PiSLF2(S2-allele of P. inflataS-locus F-box gene), was the only predicted gene whose deduced amino acid sequence was similar to the sequences of known proteins in the database. Two different cDNA selection methods were used to identify additional genes in the 881-kb contig; 11 groups of cDNA clones were identified in addition to those for S2-RNase and PiSLF2. RT-PCR analysis of expression profiles and PCR analysis of BAC clones and genomic DNA confirmed that seven of these 11 newly identified genes were located in the 881-kb contig.

cDNA selection chromosome walking Petunia inflata self-incompatibility S-locus contigs 


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Copyright information

© Kluwer Academic Publishers 2004

Authors and Affiliations

  • Yan Wang
    • 1
  • Tatsuya Tsukamoto
    • 2
  • Ki-wan Yi
    • 2
  • Xi Wang
    • 2
    • 3
  • Shihshieh Huang
    • 4
  • Andrew G. McCubbin
    • 2
    • 5
  • Teh-hui Kao
    • 1
    • 2
  1. 1.Intercollege Graduate Degree Program in Plant PhysiologyThe Pennsylvania State UniversityUniversity ParkUSA
  2. 2.Department of Biochemistry and Molecular BiologyThe Pennsylvania State UniversityUniversity ParkUSA
  3. 3.Department of Molecular, Cellular and Developmental BiologyUniversity of MichiganAnn ArborUSA
  4. 4.Mystic ResearchMonsanto CompanyMysticUSA
  5. 5.School of Biological SciencesWashington State UniversityPullmanUSA

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