甘蔗Cu/Zn-SOD的克隆和表达分析  

Molecular Cloning of Sugarcane Cu/Zn Superoxide Dismutase (Cu/Zn-SOD) and Its Expression Analysis

作　　者：王盛[1] 张保青[1] 黄杏[1,2] 樊艳姣 杨丽涛[1] 李杨瑞[1,2] 

机构地区：[1]广西大学农学院/广西大学亚热带农业生物资源保护与利用国家重点实验室,南宁530004 [2]中国农业科学院甘蔗研究中心/广西农业科学院甘蔗研究所/农业部广西甘蔗生物技术与遗传改良重点实验室/广西作物遗传改良生物技术重点开放实验室/广西甘蔗遗传改良重点实验室,南宁530007

出　　处：《中国农业科学》2013年第15期3277-3284,共8页Scientia Agricultura Sinica

基　　金：国家"863"计划课题(2013AA102604);国家国际合作项目(2009DFA30820;2013DFA31600);广西自然科学基金创新团队项目(2011GXNSFF018002);广西自然科学基金重点项目(2012GXNSFDA053011);广西自然科学基金(2013GXNSFAA019073);广西科学研究与技术开发计划项目(桂科攻1222009-1B);广西农业科学院团队项目(桂农科2011YT01)

摘　　要：【目的】克隆甘蔗铜/锌超氧化物歧化酶(Cu/Zn-SOD)基因,并分析其序列特征、组织特异性表达及在不同逆境胁迫下的表达模式。【方法】以甘蔗桂糖28号为材料,利用RT-PCR技术克隆获得Cu/Zn-SOD,采用生物信息学方法分析所推测的氨基酸序列,通过实时荧光定量PCR研究Cu/Zn-SOD在不同组织及逆境条件下的表达情况。【结果】克隆获得甘蔗Cu/Zn-SOD,NCBI登录号为JQ958328,其开放阅读框为456 bp,编码151个氨基酸,与禾本科植物聚于同一进化分支。实时荧光定量PCR分析表明Cu/Zn-SOD在根、茎、叶中均有表达,为组成型表达,在叶中表达量最高;Cu/Zn-SOD在聚乙二醇(PEG)、NaCl、低温(4℃)和H2O2四种非生物胁迫下均诱导表达,表达模式因调控机制的不同而异。【结论】克隆获得Cu/Zn-SOD,其主要在甘蔗绿色组织中表达,其在铜/锌SOD超氧化物歧化酶的功能区域保守型很高,可能与甘蔗抵御渗透胁迫相关。【Objective】The aim of this study was to clone the full-length cDNA of a key enzyme gene Cu/Zn-SOD related to superoxide anion removal in sugarcane（Saccharum spp.）,investigate its sequence characteristics and analyze its expression in different organs and under different stresses.【Method】 The Cu/Zn-SOD gene cDNA sequence was cloned from the leaves of sugarcane variety GT28 using RT-PCR techniques.The bioinformatics methods were used to analyze the putative amino acid sequence,and real-time fluorescence quantitative PCR（qRT-PCR） method was used to study the expression of Cu/Zn-SOD gene in different tissues and under different stresses.【Result】 The full-length cDNA of Cu/Zn-SOD（GenBank accession number： JQ958328） in sugarcane was cloned.The sequence included an open reading frame of 456 bp,encoding a polypeptide of 151 amino acids,which was clustered in the same phylogenetic branch of the Cu/Zn-SODs of gramineae.Real-time fluorescence quantitative PCR results showed that the Cu/Zn-SOD expressed in root,stem and leaf of sugarcane plant,belonging to constitutive expression,but showing the highest in leaf.The Cu/Zn-SOD expression could be induced under the treatments of polyethylene glycol（PEG）,H2O2,low temperature and NaCl stresses,but the expression models were different depending on the regulation mechanisms.【Conclusion】 The gene Cu/Zn-SOD was cloned,and it mainly expressed in the green tissue in sugarcane plant.Its copperzinc superoxide dismutase functional areas are highly conservative,and is possibly associated with osmotic stress resistance of sugarcane.

关 键 词：甘蔗 铜 锌超氧化物歧化酶 逆境 表达分析 实时荧光定量PCR 

分 类 号：S566.1[农业科学—作物学]