S genotyping in Japanese plum and sweet cherry by allele-specific hybridization using streptavidin-coated magnetic beads

Chun Lei Wang, Zhi Ping Zhang, Kaoru Tonosaki, Hiroyasu Kitashiba, Takeshi Nishio

Research output: Contribution to journalArticlepeer-review

2 Citations (Scopus)

Abstract

Key message: We report a rapid and reliable method forSgenotyping of Rosaceae fruit trees, which would to be useful for successful planting of cross-compatible cultivars in orchards. Japanese plum (Prunus salicina) and sweet cherry (Prunus avium), belonging to the family Rosaceae, possess gametophytic self-incompatibility controlled by a single polymorphic locus containing at least two linked genes, S-RNase and SFB (S-haplotype-specific F-box gene). For successful planting of cross-compatible cultivars of Rosaceae fruit trees in commercial orchards, it is necessary to obtain information on S genotypes of cultivars. Recently, a method of dot-blot analysis utilizing allele-specific oligonucleotides having sequences of SFB-HVa region has been developed for identification of S haplotypes in Japanese plum and sweet cherry. However, dot-blot hybridization requires considerable time and skill for analysis even of a small number of plant samples. Thus, a quick and efficient method for S genotyping was developed in this study. In this method, instead of a nylon membrane used for dot-blot hybridization, streptavidin-coated magnetic beads are used to immobilize PCR products, which are hybridized with allele-specific oligonucleotide probes. Our improved method allowed us to identify 10 S haplotypes (S-a, S-b, S-c, S-d, S-e, S-f, S-h, S-k, S-7 and S-10) of 13 Japanese plum cultivars and 10 S haplotypes (S-1, S-2, S-3, S-4, S-4′, S-5, S-6, S-7, S-9 and S-16) of 13 sweet cherry cultivars utilizing SFB or S-RNase gene polymorphism. This method would be suitable for identification of S genotypes of a small number of plant samples.

Original languageEnglish
Pages (from-to)567-576
Number of pages10
JournalPlant Cell Reports
Volume32
Issue number4
DOIs
Publication statusPublished - 2013 Apr

Keywords

  • Allele-specific oligonucleotide
  • Japanese plum
  • Magnetic bead hybridization
  • S-RNase
  • SFB
  • Sweet cherry

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