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程西永,呼晓贤,李海霞,董中东,任 妍,陈树林,詹克慧,许海霞.低温胁迫下小麦幼穗中差异表达蛋白质的鉴定[J].麦类作物学报,2016,36(10):1342
低温胁迫下小麦幼穗中差异表达蛋白质的鉴定
Identification and Analysis of Differentially Expressed Proteins in Wheat Young Spike under Cold Stress
  
DOI:10.7606/j.issn.1009-1041.2016.10.11
中文关键词:  小麦  幼穗  低温胁迫  双向电泳  胁迫响应蛋白
英文关键词:Wheat  Young spike  Low-temperature stress  Two-dimensional electrophoresis  Stress response protein
基金项目:国家重点基础研究发展计划(973计划)项目(2014CB138105);河南省教育厅重点项目(15A180039)
作者单位
程西永,呼晓贤,李海霞,董中东,任 妍,陈树林,詹克慧,许海霞 (河南农业大学/小麦玉米作物学国家重点实验室/河南粮食作物协同创新中心, 河南郑州 450002) 
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中文摘要:
      为探究低温胁迫下小麦幼穗中胁迫响应差异表达蛋白,采用IEF/SDS-PAGE双向凝胶电泳及质谱技术,以低温敏感品种SW601和低温不敏感品种陇麦157为材料,对常温生长、0 ℃低温胁迫12 h和72 h 的幼穗全蛋白进行了分析。结果表明,在pH 4~7时,SW601和陇麦157各处理幼穗蛋白图谱中均可重复检测到800~900个有效蛋白质点;低温胁迫12 h、72 h后,SW601和陇麦157中上调表达的蛋白点分别有120个和101个,下调表达的蛋白点分别有92个和60个。对2种胁迫处理下均差异表达且Ratio>2的54个蛋白点进行质谱分析,成功鉴定出43个差异蛋白。经数据库搜索匹配和蛋白质鉴定,这43个差异蛋白分别属于胁迫应激/防御相关蛋白、光合作用蛋白、蛋白代谢相关蛋白、碳水化合物代谢相关蛋白、信号传导相关蛋白、能量产生和运输相关蛋白和未知功能蛋白等7类蛋白。其中,抗坏血酸过氧化物酶、Cu/Zn超氧化物歧化酶和胚胎后期发育富集(LEA)同源蛋白14-A等蛋白的表达量在低温不敏感材料陇麦157中变化显著,而在低温敏感材料SW601中没有表达,这可能与小麦幼穗在低温逆境下的代谢调控和低温敏感特性有一定关系。
英文摘要:
      In order to study the proteins responsing to low temperature(LT) in wheat(Triticum aestivum L.) spike, two varieties SW601(LT sensitive ) and Longmai 157(LT insensitive) were used to analyze the total protein profile under low temperature at 0 ℃ for 12 h and 72 h by two-dimensional electrophoresis and matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry method,the spike under normal growth temperature conditions was control. The results show that there are 800 to 900 protein spots for each variety within pH 4 to 7. After 12 h and 72 h low temperature stress treatment,120 up-regulated and 92 down-regulated protein spots were found in SW601, while 101 up-regulated and 60 down-regulated protein spots were detected in Longmai 157. Fifty-four proteins spots which were significantly expressed with the expression ratio more than 2 under two temperature stress conditions were selected. There were 43 protein spots identified by mass spectrometry analysis.Using protein blast methods, the identified cold stress responsive proteins were classified into seven groups including stress/defense, photosynthesis, protein metabolism, carbohydrate metabolism, signal transduction, energy pathways and other unknown function. Some of the expressed proteins were enhanced significantly in LT insensitive varieties Longmai 157,such as ascorbate peroxidase,Cu/Zn superoxide dismutase,and late embryogenesis abundant,while they were not expressed in LT sensitive SW601.These protein may related to metabolic regulation or response characteristics of wheat spike under low temperature stress.
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