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.
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Takasaki, T., Okada, K., Castillo, C. et al. Sequence of the S 9-RNase cDNA and PCR-RFLP system for discriminating S 1- to S 9-allele in Japanese pear. Euphytica 135, 157–167 (2004). https://doi.org/10.1023/B:EUPH.0000014907.50575.d0
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DOI: https://doi.org/10.1023/B:EUPH.0000014907.50575.d0