新疆桃花柱S-RNase和花粉SFB基因的克隆与分析  

Cloning and analysis of S-RNase and SFBgenes in Xinjiang peach(Prunus ferganensis Kost.et Riab)

作　　者：陈秋菊[1] 孟冬[1] 李威[1] 顾钊宇[1] 段续伟[1] 袁晖[1] 张懿[1] 李天忠[1] 

机构地区：[1]中国农业大学农学与生物技术学院,北京100193

出　　处：《中国农业大学学报》2015年第6期76-83,共8页Journal of China Agricultural University

基　　金：国家自然科学基金资助项目(31372035;31171941)

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。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 SFBgenes in pollen,we designed primers to identify their S-genotypes using DNA as templates.S-RNase genes and SFBgenes 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＇S1S2respectively.The results showed that：The 602 th base guanine（G）of S2m-RNase gene was replaced by adenine（A）resulting in cysteine into tyrosine;SFB1m of S1 haplotype encoded truncated SFB1 m due to a155 bp insertion after the 978 th base.Translation of SFB2 m of S2 and S2 mhaplotypes were stopped at the 525 th base due to a 5bp insertion after the 492 th 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 SFB1 mand 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.

关 键 词：新疆桃 自交亲和性 花柱S-RNase 花粉SFB 克隆与分析 

分 类 号：S662.1[农业科学—果树学]