Identification of S haplotypes in Brassica by dot-blot analysis of SP11 alleles

R. Fujimoto, T. Nishio

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

A self-incompatibility system is used for F1 hybrid breeding in Brassicaceae vegetables. The determinants of recognition specificity of self-incompatibility in Brassica are SRK in the stigma and SP11/SCR in the pollen. Nucleotide sequences of SP11 alleles are more highly variable than those of SRK. We analyzed the S haplotype specificity of SP11 DNA by Southern-blot analysis and dot-blot analysis using 16 S haplotypes in Brassica oleracea, and found that DNA fragments of a mature protein region of SP11 cDNA, SP11m, of eight S haplotypes can detect only the SP11 alleles of the same S haplotypes. This specificity makes these methods useful for S haplotype identification. Therefore, we developed two methods of dot-blot analysis for SP11. One is dot blotting of DNA samples, i.e. plant genomic DNA probed with labeled SP11m, and the other is dot blotting of SP11m DNA fragments probed with labeled DNA samples, i.e. the SP11 coding region labeled by PCR using a template of plant genomic DNA. The former is useful for testing many plant materials. The latter is suitable, if there is no previous information on the S haplotypes of plant materials.

Original languageEnglish
Pages (from-to)1433-1437
Number of pages5
JournalTheoretical and Applied Genetics
Volume106
Issue number8
DOIs
Publication statusPublished - 2003 May 1

Keywords

  • Brassicaceae vegetables
  • Dot-blot analysis
  • F hybrid breeding
  • SP11/SCR
  • Seed purity test
  • Self-incompatibility

ASJC Scopus subject areas

  • Biotechnology
  • Agronomy and Crop Science
  • Genetics

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