水分胁迫条件下“洛旱2号”小麦根系的基因表达谱  

Gene Expression Profiling in Roots of Wheat Cultivar "Luohan 2" under Water Stress

作　　者：李永春[1] 孟凡荣[2] 王潇[1] 陈雷[1] 任江萍[1] 牛洪斌[1] 李磊[1] 尹钧[1] 

机构地区：[1]河南农业大学国家小麦工程技术研究中心,河南郑州450002 [2]河南农业大学生命科学学院,河南郑州450002

出　　处：《作物学报》2008年第12期2126-2133,共8页Acta Agronomica Sinica

基　　金：河南省科技厅项目(0623011600);国家"十一五"科技支撑计划重大项目(2006BAD02A07-4)

摘　　要：为探讨小麦水分胁迫反应的分子调控机制,以抗旱小麦品种"洛旱2号"根系为材料,采用Affymetrix小麦基因芯片比较了20%PEG6000溶液处理后根系及对照的基因表达谱。结果表明,洛旱2号根系中受水分胁迫诱导而上调和下调表达的基因分别有3743个(4074个探针组)和4573个(5043个探针组),下调表达的基因远远多于上调表达的基因,其中上调2倍以上的1593个(1716个探针组),下调2倍以上的2238个(2451个探针组)。通过Affymetrix的Net Affx Analysis Center查询和NCBI的BLASTx分析,差异表达2倍以上的基因中有619个已注释了功能,利用MIPS数据库将注释基因分为10类,包括生物代谢、逆境胁迫反应、细胞组分生成、蛋白合成、细胞信号交流、细胞运输、转录相关、细胞分裂和蛋白质加工等。差异表达16倍以上的基因中有32个已注释了功能,其中与逆境胁迫反应相关的基因16个,与生物代谢相关的基因5个。利用实时定量RT-PCR对8个代表性候选基因在水分胁迫条件下的差异表达特性分析表明,其结果和芯片杂交分析的结果基本一致。High-through microarray technology is a powerful tool for analyzing gene expression profiles of plants exposed to abiotic stresses. To investigate the molecular mechanisms of water-stress response in wheat （Triticum aestivum L.）, the transcript profiles in 20% PEG6000 treated roots and control roots of a wheat cultivar ＂Luohan 2＂ were comparatively studied using the Affymetrix wheat GeneChip. Analysis of the microarray data showed that there were 3 743 genes （4 074 probe sets） induced and 4 573 genes （5 043 probe sets） repressed in roots under water-stress. Generally, the down-regulated genes were greater in quantity than up-regulated genes. Among these differentially expressed genes, 1 593 and 2 238 genes （1 716 and 2 451 probe sets） were up- and down-regulated exceeding 2 folds, respectively. Through the sourcing annotations directly from Affymetrix website （www.affymetrix.com/analysis/） in tandem with BLASTx analyzing using the Affymetrix probe set sequences, 619 candidate genes were annotated. The functional classification of annotated genes were performed based on literature searches of the functional categories described in the MIPS database and the 619 annotated genes were classified into 10 functional categories, in- volving in metabolism, stress response, biogenesis of cellular components, protein synthesis, cellular communication, cellular transport, transcription, cell division, protein processing, and others. Thirty-two up- or down-regulated genes with over 16-fold change ratios were annotated. A half of them belonged to the stress response category, and another five were associated with metabolism. In addition, the differential expression patterns of eight representative candidate genes were confirmed by real-time quantitative RT-PCR and the results showed that the expression changes of these candidates were generally consistent with the microarray results detected by Affymetrix GeneChip.

关 键 词：水分胁迫 洛旱2号 基因差异表达谱 实时定量RT-PCR 基因芯片 

分 类 号：S512.1[农业科学—作物学]