Simple and efficient methods for S genotyping and S screening in genus Brassica by dot-blot analysis

Eriko Oikawa, Shohei Takuno, Atsushi Izumita, Koji Sakamoto, Hideaki Hanzawa, Hiroyasu Kitashiba, Takeshi Nishio

Research output: Contribution to journalArticlepeer-review

9 Citations (Scopus)

Abstract

In F1 hybrid breeding of Brassica vegetables utilizing the self-incompatibility system, identification of S genotypes in breeding lines is required. In the present study, we developed S-tester lines of 87 S haplotypes, i. e., 42 S haplotypes in B. rapa and 45 S haplotypes in B. oleracea. With these materials, we established a simple, efficient, and reliable dot-blot technique for S genotyping for 40 S haplotypes of B. rapa and and 33 of B. oleracea using allele-specific oligonucleotide probes and allele-specific primer pairs designed from sequences of each SP11 allele. In this method, DNA fragments amplified using multiplex primer pairs with digoxigenin-dUTP were hybridized with dot-blotted allele-specific oligonucleotide probes with distinct signals. In addition, we developed a screening method for identification of plants harboring a particular S haplotype using a labeled allele-specific oligonucleotide probe. This method is considered to be useful for purity testing of F1 hybrid seeds.

Original languageEnglish
Pages (from-to)1-12
Number of pages12
JournalMolecular Breeding
Volume28
Issue number1
DOIs
Publication statusPublished - 2011 Jun 1

Keywords

  • Brassica
  • Dot-blot
  • S genotyping
  • S screening
  • SP11
  • Self-incompatibility

ASJC Scopus subject areas

  • Biotechnology
  • Molecular Biology
  • Agronomy and Crop Science
  • Genetics
  • Plant Science

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