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作物学报 ›› 2012, Vol. 38 ›› Issue (09): 1561-1569.doi: 10.3724/SP.J.1006.2012.01561

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

拟南芥RBCS-1A基因受光调节表达模式及其启动子遗传转化应用评价

习雨琳1,2，周朋1，宋梅芳1，李志勇1,3，孟凡华1，杨建平1,4,*   

  1. 1 中国农业科学院作物科学研究所, 北京 100081; 2 中国农业科学院研究生院, 北京 100081; 3 河南农业大学农学院, 河南郑州 450002; 4 重庆邮电大学生物信息学院, 重庆 400065
  • 收稿日期:2012-03-06 修回日期:2012-04-20 出版日期:2012-09-12 网络出版日期:2012-07-03
  • 通讯作者: 建平, E-mail: yangjianping@caas.net.cn, Tel: 010-82105859 杨建平, E-mail: yangjianping@caas.net.cn, Tel: 010-82105859
  • 基金资助:

    本研究由国家转基因生物新品种培育重大专项(2011ZX08010-002)和重庆市自然科学基金(CSTC2009BA1088)项目资助。

Expression Pattern of Arabidopsis RBCS-1A Gene in Response to Light Treatments and Application Evaluation of Its Promoter in Transgenic Engineering

XI Yu-Lin1,2,ZHOU Peng1,SONG Mei-Fang1,LI Zhi-Yong1,3,MENG Fan-Hua1,YANG Jian-Ping1,4,*   

  1. 1 Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China; 2 Graduate School, Chinese Academy of Agricultural Sciences, Beijing 100081, China; 3 Agronomy College, Henan Agricultural University, Zhengzhou 450002, China; 4 College of Bio-information, Chongqing University of Posts and Tele-communication, Chongqing 400065, China
  • Received:2012-03-06 Revised:2012-04-20 Published:2012-09-12 Published online:2012-07-03
  • Contact: 杨建平, E-mail: yangjianping@caas.net.cn, Tel: 010-82105859

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被引次数

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摘要： RBCS编码光合碳同化关键酶核酮糖1,5-二磷酸羧化酶/加氧酶的小亚基, 是控制植物光合作用的重要基因之一。本研究利用实时荧光定量PCR技术分析拟南芥RBCS-1A受光调节表达模式, 结果表明, AtRBCS-1A表达受光诱导, 同时具有组织表达特异性; 运用生物信息学手段分析发现, 该基因启动子序列中存在多个参与光应答的顺式作用元件; 采用PCR技术从拟南芥基因组中分离到长度为1 691 bp的AtRBCS-1A启动子片段, 将该片段与GUS报告基因融合构建植物表达载体并转化野生型拟南芥, 对获得的转基因植株进行GUS染色, 结果显示, AtRBCS-1A启动子是光诱导型和组织特异型启动子。以上结果初步证明, AtRBCS-1A启动子应用于植物遗传转化切实可行, 具有重要应用价值。

关键词: 拟南芥, RBCS-1A基因, 表达模式, 光诱导表达, 顺式作用元件

Abstract: RBCS gene encodes the small subunit of Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco), which catalyzes the rate-limiting step of CO2 fixation in photosynthesis. It is very important for crop improvement to study the gene expression pattern and promoter activity of RBCS in response to light. In this study, we analyzed the gene expression pattern of Arabidopsis RBCS-1A in response to light by Real-time quantitative PCR. The results indicated that the expression of AtRBCS-1A was induced by light, and had the tissue-specific feature. In silico analysis revealed that many cis-acting elements for light regulation were found in the promoter of AtRBCS-1A. Based on this information, we isolated a genomic sequence with 1 691 bp upstream of the translation start codon of AtRBCS-1A using PCR, constructed plasmid vector with AtRBCS-1A promoter fused with GUS (encoding β-glucuronidase) reporter gene, and then generated transgenic Arabidopsis by floral dip. Histochemical analysis of the transgenic lines suggested that AtRBCS-1A promoter was a light-induced and tissue-specific promoter. All the analyses preliminarily prove that AtRBCS-1A promoter has important application value and can be used in plant transgenic engineering.

Key words: Arabidopsis, RBCS-1A gene, Expression pattern, Light-induced expression, cis-acting elements

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