首页 | 官方网站   微博 | 高级检索  
     

白菜型冬油菜铁超氧化物歧化酶(Fe-SOD) 基因的克隆及表达分析
引用本文:曾秀存,孙万仓,孙佳,许耀照,方彦,史鹏辉,杨刚,孔德晶,武军艳,刘自刚.白菜型冬油菜铁超氧化物歧化酶(Fe-SOD) 基因的克隆及表达分析[J].中国农业科学,2013,46(21):4603-4611.
作者姓名:曾秀存  孙万仓  孙佳  许耀照  方彦  史鹏辉  杨刚  孔德晶  武军艳  刘自刚
基金项目:国家“863”高技术研究发展计划(2011AA10A104)、国家公益性行业(农业)科研专项(200903002-04)、现代农业产业技术体系建设专项(CARS-13)、河西学院校长科研基金项目计划(XZ2011-02)
摘    要:【目的】了解SOD酶蛋白家族Fe-SOD在超强抗寒冬油菜中的表达情况及其在低温胁迫下的作用。【方法】采用RT-PCR技术克隆超强抗寒白菜型冬油菜陇油7号Fe-SOD的cDNA序列。对该序列进行生物信息学分析,采用半定量以及相对定量RT-PCR研究Fe-SOD在低温胁迫下的表达模式,采用氮蓝四唑(NBT)光还原法测定冬油菜叶片和根中的总SOD酶活性。【结果】获得Fe-SOD,GenBank登录号为KF178713,该基因cDNA片段全长645 bp,包含一个639 bp完整的开放阅读框,编码212个氨基酸的蛋白质,与白菜(Chiifu)的蛋白质氨基酸序列同源性高达99%。该基因编码的酶蛋白是1个主要由α-螺旋组成的亲水性稳定蛋白,无信号肽,无跨膜结构域。Fe-SOD表达模式分析显示初期低温胁迫下(4℃),该基因上调表达,继续低温胁迫处理(-4℃和-8℃),Fe-SOD表达量受到抑制。总SOD酶活性测定显示冬油菜根中酶活性高于叶片,以利于冬油菜安全越冬。【结论】从白菜型冬油菜克隆的Fe-SOD具有已知物种Fe-SOD的特征。Fe-SOD在冬油菜品种陇油7号抗寒过程中起作用。

关 键 词:白菜型油菜    低温    Fe-SOD    表达分析    SOD活性
收稿时间:2013-06-18

Cloning and Expression Analysis of Fe Superoxide Dismutase (Fe-SOD) Gene from Winter Turnip Rape (Brassica rapa L.)
ZENG Xiu-Cun-,SUN Wan-Cang-,SUN Jia-,XU Yao-Zhao-,FANG Yan-,SHI Peng-Hui-,YANG Gang-,KONG De-Jing-,WU Jun-Yan-,LIU Zi-Gang-.Cloning and Expression Analysis of Fe Superoxide Dismutase (Fe-SOD) Gene from Winter Turnip Rape (Brassica rapa L.)[J].Scientia Agricultura Sinica,2013,46(21):4603-4611.
Authors:ZENG Xiu-Cun-  SUN Wan-Cang-  SUN Jia-  XU Yao-Zhao-  FANG Yan-  SHI Peng-Hui-  YANG Gang-  KONG De-Jing-  WU Jun-Yan-  LIU Zi-Gang-
Affiliation:1.Agricultural College, Gansu Agricultural University, Lanzhou 730070, China ; 2.College of Agronomy and Biotechnology, Hexi University, Zhangye 734000, Gansu, China; 3.Department of Plant Sciences, University of Saskatchewan, Saskatoon, Saskatchewan,  Canada S7N5A8
Abstract:【Objective】The objectives of the present study were to clone the Fe-SOD gene of SOD family from an extremely low temperature (-32℃) resistant winter turnip rape (B. rapa L.) cultivar Longyou 7 and analyze its expression under low temperature conditions .【Method】The cDNA sequence of Fe-SOD was isolated by RT-PCR, and the obtained cDNA sequence and the deduced amino acid sequence was analyzed. Semi-quantitative and real time RT-PCR were used to assess the expression of Fe-SOD in response to low temperature stress. The superoxide dismutase enzyme activity was measured by NBT deoxidization method in leaves and roots. 【Result】The Fe-SOD gene was isolated from winter turnip rape (GenBank accession number. KF178713). The cDNA sequence of this gene was 645 bp in length, containing a 639 bp opening reading frame (ORF) , encoded a polypeptide of 212 amino acid,and with 99 % of amino sequence similarity to a vegetable cultivar of B. rapa (Chiifu). The protein encoded by this gene was a hydrophilic protein without signal-peptide and transmembrane region. The prediction of the second structures indicated that the Fe-SOD was a steady protein with more α-helices. Semi-quantitative and real time RT-PCR result showed that Fe-SOD was expressed upregulatedly in response to early low temperature stress (4℃). However, the expression of this gene was inhibited at the lower temperature stress (-4℃ and -8℃). The result of measured enzyme activity showed that superoxide dismutase enzyme activity in roots was higher than that of leaves,which could make winter turnip rape successfully overwinter. 【Conclusion】The Fe-SOD gene cloned from winter turnip rape had genetic characteristics similar with other known species and it might play a role in cold tolerance of the B. rapa cultivar Longyou 7.
Keywords:winter rape (Brassica rapa L  )  lower temperature  Fe-SOD  expression analysis  activity of SOD
点击此处可从《中国农业科学》浏览原始摘要信息
点击此处可从《中国农业科学》下载全文
设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司    京ICP备09084417号-23

京公网安备 11010802026262号