title: S genotyping in Japanese plum and sweet cherry by allele-specific hybridization using streptavidin-coated magnetic beads.
abstract: We report a rapid and reliable method for S genotyping 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.
authors: Graduate School of Agricultural Science, Tohoku University, 1-1 Tsutsumidori-Amamiyamachi, Aoba-ku, Sendai, Miyagi, 981-8555, Japan.
authors: Chun-Lei
authors: CL
authors: Wang
authors: Zhi-Ping
authors: ZP
authors: Zhang
authors: Kaoru
authors: K
authors: Tonosaki
authors: Hiroyasu
authors: H
authors: Kitashiba
authors: Takeshi
authors: T
authors: Nishio
mesh_terms: D000483:Alleles; D001947:Breeding; D004797:Enzyme-Linked Immunosorbent Assay; D018506:Gene Expression Regulation, Plant; D060005:Genotyping Techniques / Q000379:methods*; D006239:Haplotypes; D009693:Nucleic Acid Hybridization / Q000379:methods*; D016133:Polymerase Chain Reaction; D027861:Prunus / Q000235:genetics*; D012260:Ribonucleases / Q000235:genetics; D060436:Self-Incompatibility in Flowering Plants; D019809:Streptavidin
chemical_list: D019809:Streptavidin; D012260:Ribonucleases; C022542:ribonuclease S