草莓FvARF5基因的克隆、生物信息学及表达分析  

Cloning,Bioinformatics and Expression Analysis of FvARF5 Gene in Strawberry

作　　者：苏丽艳[1] SU Liyan(Research Center for Qinling Wild Ornamental Plant,School of Biological and Environmental Engineering,Xi′anUniversity,Xi′an 710065,China)

机构地区：[1]西安文理学院生物与环境工程学院秦岭野生观赏植物研究中心

出　　处：《华北农学报》2019年第3期16-22,共7页Acta Agriculturae Boreali-Sinica

基　　金：国家自然科学基金项目(31701935);陕西省自然科学基础研究计划项目(2017JQ3009);陕西省教育厅自然科学专项(17JK1126);西安市科技计划项目(2016CXWL03)

摘　　要：生长素响应因子ARF是一类新的转录因子,通过激活或抑制生长素响应基因的表达调控植物的生长发育过程。为解析草莓生长素响应因子FvARF5在草莓形成过程中的功能,以草莓为试验材料,克隆了FvARF5基因并进行了生物信息学分析,利用qPCR分析了FvARF5在草莓不同组织及植物激素处理下的表达模式。结果表明,FvARF5基因开放阅读框为2745bp,编码914个氨基酸,蛋白质分子质量为100.65ku,等电点为5.25。多序列比对和系统进化树分析表明,FvARF5基因具有保守的B3DNA结合结构域和生长素响应因子保守结构域,与蔷薇科月季ARF5基因的进化关系最近,相似度达95%。亚细胞定位预测分析表明,FvARF5蛋白定位于细胞核中。启动子分析显示,FvARF5基因具有多样化的激素应答元件。qPCR结果表明,FvARF5存在组织表达差异,在茎和绿果中表达量较高。另外,FvARF5基因表达受到生长素和赤霉素的诱导,初步推测该基因可能参与了生长素和赤霉素调控草莓果实的形成过程。为进一步研究FvARF5基因的功能奠定基础。Auxin response factor ARF is a new class of transcription factors. They play an important role during plant development as an activation or inhibition in auxin-related gene expression. In order to identify the function of FvARF5 gene during strawberry development, FvARF5 gene was cloned by RT-PCR from Fragaria vesca and analyzed by bioinformatics. Furthermore, Real-time quantitative PCR(qPCR)assay was performed to investigate the tissue-specific expression of FvARF5 gene,and its expression patterns under various phytohormone treatments. The results showed that the open reading frame of FvARF5 gene was 2 745 bp,encoding a protein of 914 amino acids. The molecular weight of FvARF5 protein was100.65 ku,and the theoretical pI was 5.25. Multiple sequence alignment and phylogenetic tree analysis showed that FvARF5 was closely related to ARF5 from Rosa chinensis with 95% identity. FvARF5 was predicated to be the auxin response gene that was located in the nucleus. Promoter analysis revealed that the sequence of FvARF5 contained multiple hormone response elements. Real-time PCR analysis showed that FvARF5 had specific expression difference in different organs,and it had a higher expression in stem and green fruit. Also, FvARF5 had significant response to exogenous IAA and GA treatment. The results showed that the FvARF5 might play an important role in auxin and GA signal transduction during strawberry development. This research provided a theoretical basis for further study of the function and molecular regulation mechanism of FvARF5 .

关 键 词：草莓 FvARF5 生长素 基因克隆 

分 类 号：Q78[生物学—分子生物学] S661.1[农业科学—果树学]