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  

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

作　　者：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] 

机构地区：[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

出　　处：《作物学报》2012年第9期1561-1569,共9页Acta Agronomica Sinica

基　　金：国家转基因生物新品种培育重大专项(2011ZX08010-002);重庆市自然科学基金(CSTC2009BA1088)项目资助

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技术分析了拟南芥RBCS-1A受光调节的表达模式,结果表明,AtRBCS-1A表达受到光的诱导,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1691bp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。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. In this study, we analyzed the transcription patterns of Arabidopsis RBCS-1A in response to light treatments using real-time quantitative PCR assays. The results indicated that the transcription abundance of AtRBCS-1A was induced by light, and had the tissue-specific feature. In silico analysis revealed that the promoter region of AtRBCS-1A keeps many cis-acfing elements for light regulation. Based on the information, we isolated a genomic frag- ment with 1 691 nucleotides upstream of the translation start codon of AtRBCS-1Aand constructed a binary vector with AtRBCS-1A promoter fused with GUS （encoding 13-glucuronidase） reporter gene, then generated Arabidopsis transgenicplants. Histochemical analysis of the transgenic lines suggested that AtRBCS-1A promoter was a light-induced and tissue-specific pro- moter. All the analyses preliminarily prove that AtRBCS-1A promoter has important application value in plant transgenic engi- neering.

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 

分 类 号：Q943[生物学—植物学] S33[农业科学—作物遗传育种]