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S genotyping and S screening utilizing SFB gene polymorphism in Japanese plum and sweet cherry by dot-blot analysis

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Abstract

Most Rosaceae fruit trees such as Japanese plum and sweet cherry have a gametophytic self-incompatibility (GSI) system controlled by a single S locus containing at least two linked genes with multiple alleles, i.e., S-RNase as a pistil determinant and SFB (S-haplotype-specific F-box gene) as a candidate for the pollen S determinant. For identification of S genotypes, many methods based on polymerase chain reaction (PCR) utilizing polymorphism in length of the S-RNase and SFB gene have been developed. In this study, we developed two dot-blot analysis methods for S-haplotype identification utilizing allele-specific oligonucleotides based on the SFB-HVa region, which has high sequence polymorphism. Dot-blotting of allele-specific oligonucleotides hybridized with digoxigenin-labeled PCR products allowed S genotyping of plants with nine S haplotypes (S-a, S-b, S-c, S-e, S-f, S-h, S-k, S-7 and S-10) in Japanese plum and ten S haplotypes (S-1, S-2, S-3, S-4, S-4′, S-5, S-6, S-7, S-9 and S-16) in sweet cherry (dot-blot-S-genotyping). In addition, dot-blotting of PCR products of SFB probed with the allele-specific oligonucleotides, occasionally utilizing competitive hybridization, was successful in screening for a desirable S haplotype in sweet cherry (dot-blot-S-screening).

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Abbreviations

GSI:

gametophytic self-incompatibility

SFB:

S-haplotype-specific F-box protein

PCR:

polymerase chain reaction

RFLP:

restriction fragment length polymorphism

HVa:

hypervariable region a

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Acknowledgements

This work was supported in part by a fund from the Dean of the Graduate School of Agricultural Science, Tohoku University.

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Author notes

  1. Kenta Shirasawa

    Present address: National Institute of Crop Science (NICS), Kannondai, Tsukuba, Ibaraki, 305-8518, Japan

Authors and Affiliations

  1. Graduate School of Agricultural Science, Tohoku University, Aoba-ku, Sendai, 981-8555, Japan

    Hiroyasu Kitashiba, Shao Ling Zhang, Kenta Shirasawa & Takeshi Nishio

  2. College of Horticulture, Nanjing Agricultural University, Nanjing, 210095, China

    Shao Ling Zhang & Jun Wu

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  1. Hiroyasu Kitashiba
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  2. Shao Ling Zhang
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  4. Kenta Shirasawa
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  5. Takeshi Nishio
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Correspondence to Hiroyasu Kitashiba.

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Kitashiba, H., Zhang, S.L., Wu, J. et al. S genotyping and S screening utilizing SFB gene polymorphism in Japanese plum and sweet cherry by dot-blot analysis. Mol Breeding 21, 339–349 (2008). https://doi.org/10.1007/s11032-007-9134-6

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  • DOI: https://doi.org/10.1007/s11032-007-9134-6

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