苹果‘金冠’AGO1基因克隆及与发育相关miRNA表达分析

为明确miRNA沉默靶基因的调控机理,克隆AGO1蛋白基因并了解其作为沉默复合物核心成员在miRNA沉默通路中的作用,本研究在分析‘金冠’苹果基因组的基础上,以‘金冠’叶片cDNA为模板,克隆获得了3 234bp AGO1基因全长,蛋白分子质量为119ku,等电点为9.41,包含2个保守的AGO蛋白特征结构域:PAZ和Piwi区,命名为MdAGO1。苹果‘金冠’不同组织MdAGO1基因及13种与发育相关miRNA实时定量PCR分析结果表明:1)MdAGO1在花和果实中表达量均高于叶片;2)miRNA在果实和花中表达量均较高,叶片相对较低。MdAGO1与13种发育相关miRNA表达一致,在苹果中MdAGO1可能与miRNA协同作用调控植物的生长发育。

Clone and expression of AGO1 gene and development-related miRNA in apple(Golden Delicious)

As the internal component of RNA induced silencing complex,ARGONAUTE(AGO) protein palys an important role in all known small RNA-directed regulatory pathways.In this study,the target cDNA clone of AGO1 was isolated from library prepared from Golden Delicious using RT-PCR technique.The cDNA was 3 234 bp,and would encode a protein of 1 078 amino acid with a molecular mass of 119 ku and theoretical isoelectric point of 9.41.The protein contained 2 conserved domains,PAZ and Piwi.Quantitative real-time PCR analysis revealed that Md-AGO1 was higher in flower and fruit than leaf.The aboundance of detected decelopmet related miRNA were positively correlated to Md-AGO1.Md-AGO1 and miRNA may regulate the plant growth and development together in apple.