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Deletion of a 236 kb region around S 4-RNase in a stylar-part mutant S sm4 -haplotype of Japanese pear

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Abstract

Japanese pear (Pyrus pyrifolia Nakai) has a gametophytic self-incompatibility (GSI) mechanism controlled by a single S-locus with multiple S-haplotypes, each of which contains separate genes that determine the allelic identity of pistil and pollen. The pistil S gene is the S-ribonuclease (S-RNase) gene, whereas good candidates for the pollen S gene are the F-box protein genes. A self-compatible (SC) cultivar, ‘Osa-Nijisseiki’, which is a bud mutant of ‘Nijisseiki’ (S 2 S 4), has a stylar-part mutant \( S^{{sm}}_{4} \)-haplotype, which lacks the S 4-RNase gene but retains the pollen S gene. To delineate the deletion breakpoint in the \( S^{{sm}}_{4} \)-haplotype, we constructed a bacterial artificial chromosome (BAC) library from an S 4-homozygote, and assembled a BAC contig of 570 kb around the S 4-RNase. Genomic PCR of DNA from S 4- and \( S^{{sm}}_{4} \)-homozygotes and the DNA sequence of the BAC contig allowed the identification of a deletion of 236 kb spanning from 48 kb upstream to 188 kb downstream of S 4-RNase. The \( S^{{sm}}_{4} \)-haplotype lacks 34 predicted open reading frames (ORFs) including the S 4-RNase and a pollen-specific F-box protein gene (termed as S 4 F-box0). Genomic PCR with a primer pair designed from the deletion junctions yielded a product specific for the \( S^{{sm}}_{4} \)-haplotype. The product could be useful as a maker for early selection of SC cultivars harboring the \( S^{{sm}}_{4} \)-haplotype.

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Abbreviations

BAC:

Bacterial artificial chromosome

SC:

Self-compatible

SI:

Self-incompatible

S-RNase :

S-ribonuclease

SFBB :

S locus F-box brothers

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Acknowledgments

We thank Go Suzuki (Osaka Kyoiku University, Japan) and Eigo Fukai (National Institute of Agrobiological Science, Japan) for technical advice on the construction of the BAC library, and Yuka Kashiwagi (Kobe University, Japan) for technical assistance on the construction of BAC contig. K.O. was supported by research fellowships of the Japan Society for the Promotion of Science for young scientists. This work was supported in part by Grants-in-Aid for Scientific Research (B) (nos. 16380028; 19380020) and (C) (no. 17580027) from the Ministry of Education, Science, Culture, Sports and Technology, Japan.

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Authors and Affiliations

  1. Graduate School of Agricultural Science, Kobe University, Kobe, Hyogo, 657-8501, Japan

    Kazuma Okada, Nozomi Tonaka, Yuki Moriya, Tetsu Nakanishi & Takeshi Takasaki-Yasuda

  2. Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka, 565-0871, Japan

    Naoko Norioka

  3. National Institute of Fruit Tree Science, Tsukuba, Ibaraki, 305-8605, Japan

    Yutaka Sawamura

  4. Niigata Agricultural Research Institute, Seiro, Niigata, 957-0111, Japan

    Tatsuya Matsumoto

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  1. Kazuma Okada
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  2. Nozomi Tonaka
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  3. Yuki Moriya
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  4. Naoko Norioka
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  5. Yutaka Sawamura
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  7. Tetsu Nakanishi
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  8. Takeshi Takasaki-Yasuda
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Correspondence to Takeshi Takasaki-Yasuda.

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Okada, K., Tonaka, N., Moriya, Y. et al. Deletion of a 236 kb region around S 4-RNase in a stylar-part mutant S sm4 -haplotype of Japanese pear. Plant Mol Biol 66, 389–400 (2008). https://doi.org/10.1007/s11103-007-9277-1

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