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为探索新疆桃自交亲和的原因及其与普通桃的分类关系,以4个新疆桃(Prunus ferganensis Kost.et Riab)品种为试材,分别在花柱S-RNase和花粉SFB基因保守区设计引物,在DNA中鉴定其S基因型,并通过全长引物分别克隆4个品种中的S-RNase和SFB全长基因。测序后经分析确定S基因型分别是:‘和田黄肉’S2S2m,‘喀什1号’S1S2,‘喀什4号’S2S2,‘黄李光’S1S2。DNAMAN软件比对结果表明:S2m-RNase的第602个碱基鸟嘌呤G变成了腺嘌呤A,导致半胱氨酸变成了酪氨酸;SFB1m在第978个碱基后插入155bp的片段,导致蛋白质翻译提前终止;SFB2m基因由于在第492个碱基处有5bp的片段插入,使翻译在525bp处终止。RT-PCR组织特异性分析发现新疆桃4个品种的S-RNase均具有花柱组织特异性表达,SFB均具有花粉组织特异性表达;酵母双杂交试验显示突变的SFB1m和SFB2m能分别与S1-RNase、S2-RNase和S2m-RNase相互作用。以上结果表明新疆桃的S基因与桃其他栽培品种一致,自交亲和性可能与S基因的突变相关,且S基因的突变类型与目前鉴定出的普通桃(Prunus persica L.)突变类型一致,该研究进一步说明新疆桃与普通桃的进化关系较近。
In order to explore the reason of selfing compatibility of Xinjiang peach and its classification relationship with common peach, four varieties of Prunus ferganensis Kost.et Riab were used to design the conserved regions of S-RNase and SFB in pollen, respectively Primers were used to identify their S genotypes in DNA. The full-length primers were used to clone the full-length S-RNase and SFB genes from 4 cultivars. After sequencing, the genotypes of S were determined as follows: ’Wada Huangshi’ S2S2m, ’Kashi 1’ S1S2, ’Kashi 4’ S2S2, ’Huangguang’ S1S2. DNAMAN software results showed that the 602th base guanine G of S2m-RNase was changed to adenine A, resulting in the conversion of cysteine to tyrosine. After the 978th base, SFB1m inserted a 155bp fragment Leading to an early termination of protein translation; the SFB2m gene was terminated at 525 bp due to the insertion of a 5 bp fragment at the 492th base. The results of RT-PCR showed that S-RNase of four cultivars of Xinjiang was characterized by style-specific expression and SFB with pollen-tissue specific expression. Yeast two-hybrid assay showed that the mutant SFB1m and SFB2m could interact with S1-RNase , S2-RNase and S2m-RNase interaction. The above results showed that the S gene of Xinjiang peach was consistent with other cultivars of peach, and the self-compatibility might be related to the mutation of S gene. The mutation type of S gene was consistent with the currently identified mutant type of Prunus persica L. This study further shows that the evolutionary relationship between Xinjiang peach and common peach is relatively close.