Sequence of the S
9-RNase cDNA and PCR-RFLP system for discriminating S
1- to S
9-allele in Japanese pear |
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| Authors: | Takeshi Takasaki Kazuma Okada Carlos Castillo Yuki Moriya Toshihiro Saito Yutaka Sawamura Naoko Norioka Shigemi Norioka Tetsu Nakanishi |
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| Institution: | (1) Faculty of Agriculture, Kobe University, 1-1 Rokkodai, Nada, Kobe, Hyogo 657-8501, Japan;(2) Graduate School of Science and Technology, Kobe University, 1-1 Rokkodai, Nada, Kobe, Hyogo 657-8501, Japan;(3) National Institute of Fruit Tree Science, 2-1 Fujimoto, Tsukuba, Ibaraki 305-8605, Japan;(4) Graduate School of Frontier Biosciences, Osaka University, 3-2 Yamadaoka, Suita, Osaka 565-0871, Japan |
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| Abstract: | A new S
9-allele was discovered in 6 Japanese pear cultivars, ‘Shinkou’, ‘Shinsei’, ‘Niitaka’, ‘Amanogawa’, ‘Nangetsu’ and ‘Nansui’.
cDNA encoding S
9-RNase, a stylar product of S
9-allele, was cloned from pistils of ‘Shinkou’ and ‘Shinsei’ by 3' and 5' RACE. The S
9-RNase gene had an open reading frame of 684 nucleotides encoding 228 amino acid residues. S
9-RNase had a hypervariable (HV) region different from S
1- to S
8-RNase and shared higher similarity (95.2%) with apple S
3-RNase than with 8 Japanese pear S-RNases (from 61.0% to 70.7%). Genomic PCR with primers ‘FTQQYQ’ and ‘anti-(I/T) IWPNV’ provided S
1- to S
9-amplicon (product), but could not discriminate the S
2 from the S
9 of ca. 1.3 kb. The S
2 and S
9 were distinguished by digestion with AflII and BstBI, respectively. The digestion with nine S-allele-specific restriction endonucleases, SfcI, AflII, PpuMI, NdeI,AlwNI, HincII, AccII, NruI and BstBI, distinguished S
1 to S
9, establishing that this PCR-RFLP system is useful for S-genotype assignments in Japanese pear harboring S
1- to S
9-allele. ‘Shinkou’, ‘Shinsei’, ‘Nangetsu’ and ‘Nansui’ assigned as S
4
S
9 were determined to be cross incompatible.
This revised version was published online in July 2006 with corrections to the Cover Date. |
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| Keywords: | Japanese pear Pyrus pyrifolia self-incompatibility S-allele S-RNase |
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