Cloning, Characterization and Chromosome Localization of Two Powdery Mildew Resistance-Related Gene Sequences from Wheat  

小麦中与白粉病抗性相关的两个新基因序列的克隆、特征分析及染色体定位(英文)

作　　者：于玲[1] 牛吉山[1] 马正强[1] 陈佩度[1] 齐莉莉[1] 刘大钧[1] 

机构地区：[1]南京农业大学农业部细胞遗传重点开放实验室

出　　处：《Acta Botanica Sinica》2002年第12期1438-1444,共7页Acta Botanica Sinica（植物学报：英文版）

基　　金：国家研究与开发专项 (J0 0_A_0 0 2 ) ;"863"高技术项目 (Z_17_0 4_0 1) ;国家杰出青年科学基金 (3 0 0 2 5 0 3 0 )~~

摘　　要：Reverse_transcription Polymerase Chain Reaction (RT_PCR) was performed using cDNAs as templates from wheat_ Haynaldia villosa 6VS/6AL translocation line and 'Yangmai 5' induced with fungus Erysiphe graminis , and degenerate primers designed based on the conserved amino acid sequences of known plant disease_resistance genes. The cDNA sequences encoding cyclophilin_like and H +_ATPase_like genes were first isolated and characterized in wheat. The putative amino acid sequences of the two clones showed that they were highly homologous to those of cyclophilin proteins and H +_ATPases isolated from other plants. Thus they were designated as Ta_Cyp and Ta_MAH . The obvious expression differences could be observed between wheat_ H. villosa 6VS/6AL translocation line and susceptible wheat cultivar 'Yangmai 5', implying that the two genes may be related with the resistance of wheat_ H. villosa 6VS/6AL translocation line to disease. Southern blot indicated that the wheat genome contained 2-3 copies of Ta_Cyp gene and one copy of the Ta_MAH gene. Chinese Spring nulli_tetrasomic line analysis located the Ta_Cyp homologous genes on wheat chromosome 6A, 6B and 6D. Southern blot using Ta_Cyp clone as a probe showed that the polymorphic bands existed among the H. villosa , amphiploid of Triticum durum _ H. villosa , wheat_ H. villosa 6VS/6AL translocation line and 'Yangmai 5', suggesting that Ta_Cyp homologies exist in wheat genome as well as on the short arm of chromosome 6V in H. villosa .根据抗病基因保守结构域设计简并性引物 ,以被白粉菌 (Erysiphegraminis)诱导后的小麦_簇毛麦 6VS/ 6AL易位系和“扬麦 5号”cDNA为模板进行反转录PCR(Reverse_transcriptionPolymeraseChainReaction ,RT_PCR)筛选 ,从小麦 (TriticumaestivumLinn .)中分离到 2个cDNA片段。蛋白质一级结构分析表明 ,它们分别与植物中已分离的cy clophilin蛋白和H+ _ATP酶高度同源 ,将小麦中这 2个基因分别定名为 :Ta_Cyp和Ta_MAH。经Northern杂交分析表明 ,这 2个基因在抗病的小麦_簇毛麦 6VS/ 6AL易位系及感病“扬麦 5号”中的表达水平有一定差异 ,因此推断这 2个基因有可能与小麦_簇毛麦 6VS/ 6AL易位系的抗病性相关。Southern杂交发现Ta_Cyp基因在小麦基因组中的拷贝数为 2 - 3个 ,Ta_MAH为单拷贝。利用中国春缺体_四体系 ,已将Ta_Cyp基因定位在小麦 6A、6B及 6D染色体上。用Ta_Cyp作探针 ,Southern杂交显示在簇毛麦、硬粒小麦_簇毛麦双二倍体、小麦_簇毛麦 6VS/ 6AL易位系与“扬麦 5号”之间表现多态 ,表明小麦基因组和簇毛麦染色体 6VS上有其同源基因。

关 键 词：CLONING wheat_  Haynaldia villosa   6VS/6AL translocation line cyclophilin gene H  +_ATPase gene 

分 类 号：S435.121[农业科学—农业昆虫与害虫防治]