利用cDNA-AFLP技术分析小麦应答低磷胁迫的特异表达基因  

Investigation Based on cDNA-AFLP Approach for Differential Expressed Genes Responding to Deficient-Pi in Wheat

作　　者：谷俊涛[1] 鲍金香[2] 王效颖[2] 郭程瑾[2] 李小娟[1] 路文静[1] 肖凯[2] 

机构地区：[1]河北农业大学生命科学学院,河北保定071001 [2]河北农业大学农学院,河北保定071001

出　　处：《作物学报》2009年第9期1597-1605,共9页Acta Agronomica Sinica

基　　金：国家重点基础研究发展计划(973计划)前期项目(2007CB116209);河北省重点基础研究项目(08965525D);河北省自然科学基金(C2007000476)资助

摘　　要：以磷高效小麦品种石新828为材料,采用cDNA-AFLP技术,鉴定了短期(1～6h)、中期(12～48h)和长期(72～144h)低磷胁迫根系特异上、下调表达基因的表达序列标签(EST)。共有非重复的上调ESTs142个,下调ESTs94个。胁迫下的前者分别含短、中和长期23、53和66个;后者分别含短、中和长期17、39和38个。对其功能比对发现,上调ESTs在功能上归属于信号转导、转录调控、代谢、逆境响应、发育、物质运输、脂类代谢和功能未知等类别,下调EST除上述类别外,还含有蛋白质合成和降解等类别。部分转录因子基因(如水稻OsPTF1和拟南芥ZAT10高度同源的转录因子基因)、促分裂原激酶基因MAPK1a、钙依赖蛋白激酶基因CPK1A和蛋白激酶基因(如serine/threonine kinase)、高亲和磷转运蛋白基因(PHT3和PT2)、过氧化物酶基因(如peroxidase73)和谷胱甘肽-S-转移酶基因(glutathione S-transferase),受到低磷胁迫的特异增强诱导,在改善小麦植株对低磷胁迫的适应能力中可能具有重要作用。研究表明,小麦对低磷胁迫的响应,在分子水平上存在着植株感受低磷胁迫信号和信号转导、进一步在生理生化方面对胁迫信号产生应答等复杂的过程。To date, differential expression genes in response to deficient-Pi stress have been identified and well studied in Arabidopsis thaliana, and totally 612 up-regulated and 254 down-regulated genes with various functions were reported. However, there is no similar report in wheat （Triticum aestivum L.）. In this study, seedlings of wheat cultivar Shixin 828 with high phosphorus use efficiency were treated with 20 μmol L^-1 Pi for 1 to 144 h, and the differential expressed sequence tags （ESTs） with up-regulated and down-regulated patterns were investigated based on cDNA-AFLP approach after deficient-Pi treatment for short term （1-6 h）, medium term （12-48 h）, and long term （72-144 h）. A total of 142 nonredundant ESTs with up-regulated pattern were identified, in which 23, 53, and 66 ESTs expressed in treatments of short, medium, and long term, respectively. Simultaneously, 94 nonredun- dant ESTs with down-regulated pattern were detected in treatments of short （17）, medium （39）, and long term （38）. These ESTs were classified into several functional groups with Blast in GenBank. Except for 44 function-unknown ESTs with the up-regulated pattern, the remained up-regulated ESTs conferred functions of signal transduction, transcription regulation, metabolism, stress response, development, transport, and lipid metabolism. Besides the above functions, protein synthesis and protein degradation were also observed in the down-regulated ESTs. Some genes of transcription factors （such as the transcription factor genes with high homologous to rice OsPTF1 and Arabidospsis ZAT10）, mitogen activated protein kinase （MAPK1a）, calcium-dependent pro- tein kinase （CPK1A）, and protein kinase （such as serine/threonine kinase）, high-affinity phosphate transporter （PHT3 and PT2）, peroxidase （such as peroxidase 73） and glutathione （glutathione S-transferase） were specifically up-regulated under deficient-Pi condition. This suggested that they might play important roles in promoting ada

关 键 词：小麦 低磷胁迫 cDNA—AFLP 特异表达基因 基因功能类别 

分 类 号：S512.1[农业科学—作物学]