草莓生长素响应因子FvARF10和FvARF18基因的克隆及表达分析

Cloning and Characterization of Auxin Response Factor Gene FvARF10 and FvARF18 in Fragaria vesca

该研究以森林草莓(Fragaria vesca)为试验材料,利用RT-PCR技术克隆草莓FvARF10、FvARF18基因全长cDNA序列。生物信息学分析表明,FvARF10、FvARF18基因开放阅读框(ORF)分别为2 056bp和2 100bp,编码685和699个氨基酸,预测其分子量和等电点分别为75.73、76.78kD和8.12、6.55。系统进化树分析表明,FvARF10与苎麻BnARF10聚为一个分支,FvARF18与桑树MnARF18聚集在一个分支上,进化关系较密切。亚细胞定位预测分析表明,FvARF10、FvARF18蛋白均定位于细胞核或细胞质中。启动子分析显示,FvARF10、FvARF18基因均具有多样化的激素应答元件。荧光定量PCR结果显示,FvARF10和FvARF18基因在草莓不同组织中均有表达,FvARF10在果实中表达较高,且在半熟果中表达量最高,而FvARF18在茎中相对表达量最高;外源激素IAA处理1h后,FvARF10和FvARF18基因受到强烈诱导均呈上调表达,而后渐恢复正常水平。研究结果表明,FvARF10和FvARF18基因可能参与草莓的营养生殖及果实的生长发育调控,且与生长素信号转导过程相关。

Two auxin response genes named FvARF10 and FvARF18 were cloned from Fragaria vesca using RT-PCR.Bioinformatics analysis showed that FvARF10 and FvARF18 contained 2 056 bp and 2 100 bp ORFs,encoding 685 and 699 amino acid residues,respectively.It is predicted that the molecular mass of FvARF10 and FvARF18 proteins were 75.73 kD and 76.78 kD,which isoelectric point were 8.12 and 6.55.Phylogenetic tree analysis indicated that FvARF18 was most closely related to MnARF18 from Morus notabilis,and FvARF10 was most closely related to BnARF10 gene from Boehmeria nivea.FvARF10 and FvARF18 are predicated to be auxin response genes that are located in the nucleus or cytoplasm.Promoter analysis revealed that the sequence of FvARF10 and FvARF18 contained multiple hormone response elements.Results of the real-time PCR analysis showed that FvARF10 and FvARF18 were constitutively expressed in all examined tissues.FvARF18 was strongly expressed in stem,while FvARF10 was strongly expressed in half red fruit.The relative transcript level of FvARF10 and FvARF18 were induced strongly under IAA treatment,and reached the maximum after treated by IAA 1 h.These results indicated that FvARF10 and FvARF18 genes may be involved in the regulation of vegetative reproduction and development of strawberry,which related to the signal transduction of auxin.