甘蔗Cu/Zn-SOD的克隆和表达分析

doi:10.3864/j.issn.0578-1752.2013.15.023

中国农业科学 ›› 2013, Vol. 46 ›› Issue (15): 3277-3284.doi: 10.3864/j.issn.0578-1752.2013.15.023

甘蔗Cu/Zn-SOD的克隆和表达分析

王盛1, 张保青1, 黄杏12, 樊艳姣1, 杨丽涛1, 李杨瑞12

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

收稿日期:2013-01-25 出版日期:2013-08-01 发布日期:2013-05-06
通讯作者: 通信作者杨丽涛，E-mail：liyr@gxu.edu.cn；通信作者李杨瑞，E-mail：liyr@gxaas.net
作者简介:王盛，E-mail：wangsheng1021@126.com
基金资助:
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）、广西自然科学基金（2013GXNSFAA019073）

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

WANG Sheng-1, ZHANG Bao-Qing-1, HUANG Xing-12, FAN Yan-Jiao-1, YANG Li-Tao-1, LI Yang-Rui-12

1.College of Agriculture, Guangxi University/State Key Laboratory for Conservation and Utilization of Subtropical Agro- Bioresources, Guangxi University, Nanning 530004
2. Sugarcane Research Center, Chinese Academy of Agricultural Sciences/ Sugarcane Research Center, Guangxi Academy of Agricultural Sciences/Key Laboratory of Sugarcane Biotechnology and Genetic Improvement (Guangxi), Ministry of Agriculture/Guangxi Crop Genetic Improvement and Biotechnology Laboratory/Guangxi Key Laboratory of Sugarcane Genetic Improvement, Nanning 530007

Received:2013-01-25 Online:2013-08-01 Published:2013-05-06

摘要/Abstract

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。

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

Abstract: 【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 copper- zinc superoxide dismutase functional areas are highly conservative, and is possibly associated with osmotic stress resistance of sugarcane.

Key words: sugarcane (Saccharum spp.) , Cu/Zn-SOD , adverse stress , expression analysis , real-time fluorescence quantitative PCR (qRT-PCR)

王盛1, 张保青1, 黄杏12, 樊艳姣1, 杨丽涛1, 李杨瑞12. 甘蔗Cu/Zn-SOD的克隆和表达分析[J]. 中国农业科学, 2013, 46(15): 3277-3284.

WANG Sheng-1, ZHANG Bao-Qing-1, HUANG Xing-12, FAN Yan-Jiao-1, YANG Li-Tao-1, LI Yang-Rui-12. Molecular Cloning of Sugarcane Cu/Zn Superoxide Dismutase (Cu/Zn-SOD) and Its Expression Analysis[J]. Scientia Agricultura Sinica, 2013, 46(15): 3277-3284.

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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

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