受条锈菌诱导的小麦丝氨酸苏氨酸激酶基因TaS/TK的克隆与表达  

Cloning and expression analysis of a Serine/Threonine protein kinase gene TaS/TK in wheat in response to stripe rust fungal infection

作　　者：蒋正宁[1] 别同德[1] 赵仁惠 高德荣[1] 吴旭江[1] 张伯桥[1] JIANG Zheng-ning BIE Tong-de ZHAO Ren-hui GAO De-rong WU Xu-jiang ZHANG Bo-qiao(Key Laboratory of Wheat Biology and Genetic Improvement on Low ＆ Middle Yangtze River Valley Winter Wheat Region, Ministry of Agriculture/Institute of Agricultural Science of the Lixiahe District in Jiangsu Province, Yangzhou 225007, Chin)

机构地区：[1]江苏里下河地区农业科学研究所/农业部长江中下游小麦生物与遗传改良重点实验室,江苏扬州225007

出　　处：《江苏农业学报》2016年第5期980-986,共7页Jiangsu Journal of Agricultural Sciences

基　　金：国家重点研发计划项目(2016YFD0101802);江苏省自然科学青年基金项目(BK20140500);江苏省农业科技自主创新基金项目[CX(14)5080]

摘　　要：以小麦基因芯片数据获得的1个受条锈病菌诱导上调表达的EST序列为探针,从小麦中克隆抗条锈病相关基因TaS/TK,并对其序列特征、进化关系和表达特征进行分析。结果表明:TaS/TK基因c DNA全长为1 582 bp,包含1 239 bp的ORF、139 bp的5'UTR以及240 bp的3'UTR,编码412个氨基酸,分子量47 120,等电点为5.84,并将其定位于小麦5B染色体;SMART软件分析结果表明TaS/TK仅存在1个典型的丝氨酸/苏氨酸蛋白激酶结构域;经激酶区Blastp比对分析,TaS/TK与其他植物同源性较低,与同源性最高的短柄草仅为59%;qRT-PCR检测结果显示,TaS/TK在小麦茎、叶中均有表达,但在根中低水平表达;TaS/TK在抗病小麦、感病小麦中均受条锈菌诱导表达,但在抗病材料中表达水平明显高于感病材料;同时,TaS/TK受水杨酸诱导上调表达。以上结果表明TaS/TK可能参与小麦SA信号通路中对条锈病菌的抗性反应。In this study, using an up-regulated EST screened by gene chip as probe, a stripe rust resistance-related gene TaS/TK was cloned from wheat,and the gene sequence features, evolutionary relationships and expression characteris-tics were analyzed. The full-length cDNA sequence of TaS/TK is 1 582 bp in length, containing a 1 239-bp open reading frame（ ORF） , a 139-bp 5′UTR and a 240-bp 3′UTR. TaS/TK was predicted to encode a protein of 412 amino acids with a molecular mass of 47 120 and an isoeletric point （ pI） of 5-84, and was located on chromosome 5B. SMART analysis re-vealed this putative protein （ TaS/TK） only possessed the activities of the serine/threonine kinase. Phylogenetic analysis showed that TaS/TK was lowly homologous to the orthologous genes from rice, maize and barley, and the highest similarity （ 59%） was to that from Brachypodium distachyon. Real time quantitative PCR exhibited low expression of TaS/TK in roots but high in stems and leaves. TaS/TK was obviously responsive to stripe rust pathogen, and presented rather higher expression in resistant materials than in susceptible materials. Furthmore, the transcription level of TaS/TK was strongly induced by salicylic acid （SA）.These results imply that TaS/TK might be involved＆amp;nbsp;in the SA-dependent stripe rust resistance responding gene network in wheat.

关 键 词：小麦 条锈病 丝氨酸苏氨酸激酶基因 实时定量PCR 

分 类 号：S512.1[农业科学—作物学]