Identification of S-alleles using polymerase chain reaction-cleaved amplified polymorphic sequence of the S-locus receptor kinase in inbreeding lines of Brassica oleracea

J. I. Park, S. S. Lee, M. Watanabe, Y. Takahata, I. S. Nou

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Identification and DNA polymorphism of the S-locus receptor kinase gene (SRK) was analysed by pollen tube tests, polymerase chain reaction-cleaved amplified polymorphic sequence (PCR-CAPS) and nucleotide sequencing. SRK-specific primers that can distinguish class I and class II S haplotypes amplified single DNA fragments of 900-1050 bp. The DNA fragments of 22 inbred lines amplified with a class I SRK-specific primer pair determined seven types with HinfI and EcoRII. In addition, the DNA fragments of 17 inbred lines amplified with a class II SRK-specific primer pair determined three types with AluI. Nucleotide sequencing of the DNA fragments amplified from 10 S haplotypes showed that exons of the 3′-end in SRK are highly conserved, and that there is much variation of the introns, which produced polymorphism of the band pattern in PCR-CAPS profiles. The S haplotypes of the plants were determined by restriction analysis of PCR products and agreed with results based on pollen tube growth tests. The PCR-CAPS analysis using specific primer pairs of SRK is considered to be useful for S allele identification in breeding programmes.

Original languageEnglish
Pages (from-to)192-197
Number of pages6
JournalPlant Breeding
Volume121
Issue number3
DOIs
Publication statusPublished - 2002 Jul 24
Externally publishedYes

Keywords

  • Brassica oleracea var. capitata
  • Brassica oleracea var. italica
  • PCR-CAPS
  • S alleles
  • S-locus receptor kinase
  • Self-incompatibility

ASJC Scopus subject areas

  • Agronomy and Crop Science
  • Genetics
  • Plant Science

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