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基于荧光共振能量转移的报告基因用于直接可视化拟南芥中脱落酸浓度变化及分布情况

FRET-based reporters for the direct visualization of abscisic acid concentration changes and distribution in Arabidopsis.

作者信息

Waadt Rainer, Hitomi Kenichi, Nishimura Noriyuki, Hitomi Chiharu, Adams Stephen R, Getzoff Elizabeth D, Schroeder Julian I

机构信息

Division of Biological Sciences, Cell and Developmental Biology Section, University of California, San Diego, La Jolla, United States.

出版信息

Elife. 2014 Apr 15;3:e01739. doi: 10.7554/eLife.01739.

DOI:10.7554/eLife.01739
PMID:24737861
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3985518/
Abstract

Abscisic acid (ABA) is a plant hormone that regulates plant growth and development and mediates abiotic stress responses. Direct cellular monitoring of dynamic ABA concentration changes in response to environmental cues is essential for understanding ABA action. We have developed ABAleons: ABA-specific optogenetic reporters that instantaneously convert the phytohormone-triggered interaction of ABA receptors with PP2C-type phosphatases to send a fluorescence resonance energy transfer (FRET) signal in response to ABA. We report the design, engineering and use of ABAleons with ABA affinities in the range of 100-600 nM to map ABA concentration changes in plant tissues with spatial and temporal resolution. High ABAleon expression can partially repress Arabidopsis ABA responses. ABAleons report ABA concentration differences in distinct cell types, ABA concentration increases in response to low humidity and NaCl in guard cells and to NaCl and osmotic stress in roots and ABA transport from the hypocotyl to the shoot and root. DOI: http://dx.doi.org/10.7554/eLife.01739.001.

摘要

脱落酸（ABA）是一种植物激素，可调节植物生长发育并介导非生物胁迫响应。直接对响应环境信号的动态ABA浓度变化进行细胞监测对于理解ABA的作用至关重要。我们开发了ABAleons：ABA特异性光遗传学报告基因，其可瞬间将植物激素触发的ABA受体与PP2C型磷酸酶的相互作用转化为响应ABA的荧光共振能量转移（FRET）信号。我们报告了亲和力在100 - 600 nM范围内的ABAleons的设计、构建及应用，以时空分辨率绘制植物组织中ABA浓度变化图。高ABAleon表达可部分抑制拟南芥的ABA响应。ABAleons可报告不同细胞类型中的ABA浓度差异、保卫细胞中响应低湿度和NaCl以及根中响应NaCl和渗透胁迫时ABA浓度的增加，以及ABA从下胚轴向地上部和根部的转运。DOI：http://dx.doi.org/10.7554/eLife.01739.001 。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4c28/3985518/25459b2ef92c/elife01739f009.jpg
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