| 摘要: |
| 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155 bp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5 bp的片段插入,使翻译在525 bp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。 |
| 关键词: 新疆桃 自交亲和性 花柱S-RNase 花粉SFB 克隆与分析 |
| DOI：10.11841/j.issn.1007-4333.2015.06.10 |
| 投稿时间：2015-01-29 |
| 基金项目:国家自然科学基金资助项目(31372035,31171941) |
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| Cloning and analysis of S-RNase and SFB genes in Xinjiang peach (Prunus ferganensis Kost.et Riab) |
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CHEN Qiu-ju, MENG Dong, LI Wei, GU Zhao-yu, DUAN Xu-wei, YUAN Hui, ZHANG Yi, LI Tian-zhong
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| (College of Agronomy and Biotechnology, China Agricultural University, Beijing 100193, China) |
| Abstract: |
| To explore the reasons of self-compatibility of Xinjiang peach and reveal its relationship with peach.Four Xinjiang cultivars were used in this study.According to the conserved domain of S-RNase genes in style and SFB genes in pollen,we designed primers to identify their S-genotypes using DNA as templates.S-RNase genes and SFB genes of four cultivars were cloned by full-length primers.After sequence alignment by DNAMAN software,we identified S-genotypes of the four cultivars.They were ‘Hetian Huang Rou’ S2S2m,‘Kashgar 1’ S1S2,‘Kashgar 4’ S2S2 and ‘Huang Li Guang’ S1S2 respectively.The results showed that:The 602th base guanine (G) of S2m-RNase gene was replaced by adenine(A) resulting in cysteine into tyrosine; SFB1m of S1 haplotype encoded truncated SFB1m due to a 155 bp insertion after the 978th base.Translation of SFB2m of S2 and S2m haplotypes were stopped at the 525th base due to a 5 bp insertion after the 492th base.Tissue-specific RT-PCR analysis showed that the S-RNase genes of four cultivars had style tissue-specific expression,whereas SFB genes were only expressed in pollen.Yeast two-hybrid experiments showed that SFB1m and SFB2m mutant interacted with S1-RNase,S2-RNase and S2m-RNase,respectively.In conclusion,S genes were the same with peach (Prunus persica L.) cultivars,and then self-compatibility might be associated with mutations of S-RNase and SFB.The mutations of S genes in Xinjiang peach were consistent with the mutations identified in peach.The study further illustrated that the evolutionary relationship between Xingjiang peach and peach (Prunus persica L.) was close. |
| Key words: Xingjiang Peach self-compatibility S-RNase SFB cloning and analysis |
