Molecular identification of a root apical cell-specific and stress-responsive enhancer from an Arabidopsis enhancer trap line

文献类型： 外文期刊

第一作者： Zhang, Lei

作者： Zhang, Lei;Qin, Li-Na;Zeng, Zi-Rui;Wu, Chang-Zheng;Gong, Yuan-Yong;Liu, Lai-Hua;Qin, Li-Na;Gong, Yuan-Yong;Cao, Feng-Qiu

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关键词： Enhancer trap line; Enhancer; Root apex specific expression; GFP and GUS; Abiotic stimuli

期刊名称：PLANT METHODS （ 影响因子：4.993； 五年影响因子：5.312 ）

ISSN： 1746-4811

年卷期： 2019 年 15 卷

页码：

收录情况： SCI

摘要： BackgroundPlant root apex is the major part to direct the root growth and development by responding to various signals/cues from internal and soil environments. To study and understand root system biology particularly at a molecular and cellular level, an Arabidopsis T-DNA insertional enhancer trap line J3411 expressing reporters (GFP) only in the root tip was adopted in this study to isolate a DNA fragment.ResultsUsing nested PCR, DNA sequencing and sequence homology search, the T-DNA insertion site(s) and its flanking genes were characterised in J3411 line. Subsequently, a 2000bp plant DNA-fragment (E-rtip1) upstream of the insert position of the coding T-DNA was in silico analysed, revealing certain putative promoter/enhancer cis-regulatory elements. Cloning and transformation of this DNA fragment and its truncated segments tagged with or without 35S minimal promoter (35Smini), all of which were fused with a GFP or GUS reporter, allowed to detect GFP and GUS expression mediated only by E-rtip1+35mini (PErtip1+35Smini) specifically in the Arabidopsis root tip region. The PErtip1+35Smini activity was further tested to be strong and stable under many different growth conditions but suppressed by cold, salt, alkaline pH and higher ammonium and phosphorus.ConclusionThis work describes a promising strategy to isolate a tissue-/cell-specific enhancer sequence from the enhancer trap lines, which are publically available. The reported synthetic promoter i.e. PErtip1+35Smini may provide a valuable and potent molecular-tool for comprehensive investigation of a gene function related to root growth and development as well as molecular engineering of root-architectural formation aiming to improve plant growth.

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