草莓FvARF2的克隆、组织表达分析及对外源激素的响应

Cloning and Tissue Expression Analysis and Response to the Exogenous Phytohormones of FvARF2 from Strawberry

ARFs家族基因广泛参与植物生长发育的调控过程。为了研究草莓生长素响应因子FvARF2在草莓生长发育过程中的功能,以‘丰香’草莓为实验材料,利用RT-PCR技术克隆草莓FvARF2基因,对其序列特征进行生物信息学分析,并利用Real-time PCR技术分析了Fv ARF2在草莓不同组织(包括根,茎,叶,花,绿果, 1/2红果及红熟果)及外源激素生长素(IAA)、乙烯利(C2H4)、脱落酸(ABA)处理下的表达模式。结果表明,Fv ARF2基因开放阅读框为2 469 bp,编码822个氨基酸;预测其分子量和等电点分别为91.1 kD和6.11,具有保守的B3 DNA结合结构域和生长素响应因子保守结构域;该基因启动子区含有生长素、ABA、茉莉酸甲酯及胁迫相关的顺式作用元件。进化分析表明FvARF2与甜樱桃ARF2B基因亲缘关系最近;Real-time PCR结果表明,FvARF2基因表达量存在组织表达差异,且对外源IAA、ABA和乙烯有显著性应答反应,说明该基因可能参与了生长素、ABA和乙烯调控草莓发育的信号转导过程。本研究为后继研究草莓FvARF2的功能和作用机制作为参考依据。

The ARFs genes widely take part in the regulating process of plant growth and development. In order to identify the function of FvARF2 gene in strawberry development. FvARF2 gene was cloned by RT-PCR from’Fengxiang’ strawberry, This study took ’Fengxiang’ strawberry as the test materials, cloned FvARF2 from strawberry by RT-PCR, and analyzed the sequence feature of Fv ARF2 gene by bioinformatics methods. And the relative expression levels of Fv ARF2 gene was analyzed by real-time quantitative PCR(qPCR) method in different tissues(root, leaf, stem, flower, green fruit, 1/2 red fruit, red fruit), and the expression of FvARF2 responding to IAA, ethephon and abscisic acid was further investigated by qPCR. The results showed that FvARF2 contained a2 469 bp open reading frame and encoded 822 amino acid. The Fv ARF2 protein molecular mass was 91.1 kD and isoelectric point was 6.11. Fv ARF2 has B3 DNA-binding and auxin response factor conserved domains. Gene promoter region contained auxin, ABA, Me JA and stress related cis-acting element. The Phylogenetic tree analysis indicated that FvARF2 has a closer relationship with Prunus avium ARF2 B. Real-time PCR analysis showed FvARF2 had specific expression differences in different organs, and it had a significant response to exogenous IAA, ABA and ethylene treatment. Those showed that the FvARF2 might play an important role in auxin, ABA,and ethylene signal transduction during strawberry development. This research might provide a theoretical basis for further study of the function and molecular regulation mechanism of FvARF2.