Sequence of the S₉-RNase cDNA and PCR-RFLP system for discriminating S₁- to S₉-allelein Japanese pear

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• 作者：Takasaki ; Takeshi ; Okada ; Kazuma ; Castillo ; Carlos ; Moriya ; Yuki ; Saito ; Toshihiro ; Sawamura ; Yutaka ; Norioka ; Naoko ; Norioka ; Shigemi ; Nakanishi ; Tetsu
• 关键词：Japanese pear ; Pyrus pyrifolia ; self ; incompatibility ; S ; allele ; S ; RNase
• 刊名：Euphytica
• 出版时间：2004
• 出版年：2004
• 期刊代码：43_00142336
• 类别：bio
• 卷：135
• 期：2
• 页码：157-167
• 数据来源：sp

摘要

A new S₉-allele was discovered in 6 Japanese pear cultivars, `Shinkou', `Shinsei', `Niitaka', `Amanogawa', `Nangetsu' and `Nansui'. cDNA encoding S₉-RNase, a stylar product of S₉-allele, was cloned from pistils of `Shinkou' and `Shinsei' by 3' and 5' RACE. The S₉-RNase gene had anopen reading frame of 684 nucleotides encoding 228 amino acid residues. S₉-RNase had a hypervariable (HV) region different from S₁- to S₈-RNase and shared higher similarity (95.2%) with apple S₃-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 S₁- to S₉-amplicon(product), but could not discriminate the S₂ from the S₉ of ca. 1.3 kb. The S₂ and S₉ were distinguished by digestion with AflIIand BstBI, respectively. The digestion with nine S-allele-specific restriction endonucleases, SfcI, AflII, PpuMI, NdeI,AlwNI, HincII, AccII, NruI and BstBI, distinguished S₁ to S₉, establishing that this PCR-RFLP system is useful for S-genotype assignments in Japanese pear harboring S₁- to S₉-allele. `Shinkou', `Shinsei', `Nangetsu' and `Nansui' assigned as S₄S₉ were determined to be cross incompatible.