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

研究成果: Article査読

9 被引用数 (Scopus)

抄録

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.

本文言語English
ページ(範囲)1-12
ページ数12
ジャーナルMolecular Breeding
28
1
DOI
出版ステータスPublished - 2011 6

ASJC Scopus subject areas

  • バイオテクノロジー
  • 分子生物学
  • 農業および作物学
  • 遺伝学
  • 植物科学

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