基于生物信息学分析的水稻无机焦磷酸化酶基因OsIP1的克隆及其遗传转化  

Cloning of an Inorganic Pyrophosphatase Gene OsIP1 in Rice Based on Bioinformatics Analysis and its Genetic Transformation

作　　者：张亚芳[1] 余永旗[1] 左示敏[1] 娄丽娟[1] 陈宗祥[1] 潘学彪[1] 

机构地区：[1]扬州大学江苏省作物遗传生理重点实验室/教育部植物功能基因组学重点实验室,江苏扬州225009

出　　处：《中国水稻科学》2013年第1期17-25,共9页Chinese Journal of Rice Science

基　　金：国家自然科学基金资助项目(30800067);中国科学院创新基金资助项目;江苏高校优势学科建设工程资助项目

摘　　要：无机焦磷酸化酶在植物体内催化焦磷酸基团分解为磷酸基团的反应,而焦磷酸的及时降解,被认为是原初光合产物合成双糖特别是蔗糖,进而进行长距离运输的关键步骤之一。但是,蔗糖在维管束中的运输需要焦磷酸,因此,在叶肉细胞中特异性过表达焦磷酸化酶基因,被认为是拉动和促进光合作用的关键措施之一。对水稻中约30个无机焦磷酸化酶编码基因进行氨基酸序列比较、结构域特征、亚细胞定位预测以及上游顺式元件释义等生物信息学分析,进而克隆了1个预测为编码胞质型可溶性无机焦磷酸化酶的基因OsIP1(Os04g0687100),并将其与叶肉细胞特异性启动子cyFBPase相连,构建成嵌合基因cyFBPase:OsIP1;通过农杆菌转化法将其转入2个水稻品种中,累计获得48个阳性转基因植株。Inorganic pyrophosphatase hydrolyzes inorganic pyrophosphate（PPi） to two ions of phosphate radical（Pi） in vivo.The timely degradation of PPi has been recognized as a key rate-limiting step in the process of disaccharide synthesis,especially sucrose.However,maintaining certain concentration of PPi in vascular bundle cells is required for the translocation of sucrose.Therefore,it could be the best resolution to over-express inorganic pyrophosphatase gene only in mesophyll tissue,which would feed back and accelerate photosynthesis and produce more carbohydrate.In this research,we firstly analyzed 30 inorganic pyrophosphatase genes predicted in rice by a series of bioinformatics softwares,such as protein clusters,amino acids sequence alignment,protein subcellular localization prediction and cis-elements analysis.Secondly,one gene（Os04g0687100） that was considered encoding soluble cytoplasmic inorganic pyrophosphatase,named as OsIP1,was cloned and placed under the rice mesophyll-specific promoter cyFBPase from a cytosolic fructose-1,6-bisphosphatase gene.Thirdly,we transformed this chimeric gene,cyFBPase：OsIP1,into two different source-sink type rice varieties using Agrobacterium-mediated method,and obtained a total of 48 independent positive transgenic plants.

关 键 词：水稻 无机焦磷酸化酶 基因克隆 叶肉细胞特异启动子 

分 类 号：S511[农业科学—作物学]