Live Cell Imaging with R-GECO1 Sheds Light on flg22- and Chitin-Induced Transient [Ca(2+)]cyt Patterns in Arabidopsis.
Live Cell Imaging with R-GECO1 Sheds Light on flg22- and Chitin-Induced Transient [Ca(2+)]cyt Patterns in Arabidopsis.
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DOI：
10.1016/j.molp.2015.05.006
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发表时间：
2015-08
期刊：
影响因子：
27.5
通讯作者：
Krebs, Melanie
中科院分区：
文献类型：
--
作者：
Keinath, Nana F.;Waadt, Rainer;Brugman, Rik;Schroeder, Julian I.;Grossmann, Guido;Schumacher, Karin;Krebs, Melanie
细胞内钙瞬变是病原相关分子模式（PAMP）触发的植物免疫过程中信号级联的一个组成部分。然而，我们对PAMP诱导的Ca 2+信号的空间分布的了解是有限的。研究细胞和组织特异性的Ca 2+依赖性信号传导过程需要多功能的Ca 2+报告者，这些报告者能够从细胞和亚细胞结构以及从整个组织中提取空间信息，时间从几秒到几小时。基于荧光的报告分子覆盖了广泛的空间和时间范围，这使得它们非常适合研究活细胞中的Ca 2+信号。在这项研究中，我们比较了两种基于荧光的Ca 2+传感器：基于Förster共振能量转移（FRET）的报告子黄色cameleon NES-YC 3.6和基于强度的传感器R-GECO 1。我们证明，R-GECO 1表现出显着增加的信号变化相比，比率NES-YC 3.6在几个刺激。由于其上级灵敏度，R-GECO 1能够在细胞尺度上报告flg 22和几丁质诱导的Ca 2+信号，这允许识别响应真菌诱导子几丁质的表皮和保卫细胞中的确定的[Ca 2 +]细胞质振荡。此外，我们发现flg 22-和几丁质诱导的根中的Ca 2+信号起始于伸长区。
Intracellular Ca2+ transients are an integral part of the signaling cascade during pathogen-associated molecular pattern (PAMP)-triggered immunity in plants. Yet, our knowledge about the spatial distribution of PAMP-induced Ca2+ signals is limited. Investigation of cell- and tissue-specific properties of Ca2+-dependent signaling processes requires versatile Ca2+ reporters that are able to extract spatial information from cellular and subcellular structures, as well as from whole tissues over time periods from seconds to hours. Fluorescence-based reporters cover both a broad spatial and temporal range, which makes them ideally suited to study Ca2+ signaling in living cells. In this study, we compared two fluorescence-based Ca2+ sensors: the Förster resonance energy transfer (FRET)-based reporter yellow cameleon NES-YC3.6 and the intensity-based sensor R-GECO1. We demonstrate that R-GECO1 exhibits a significantly increased signal change compared with ratiometric NES-YC3.6 in response to several stimuli. Due to its superior sensitivity, R-GECO1 is able to report flg22- and chitin-induced Ca2+ signals on a cellular scale, which allowed identification of defined [Ca2+]cyt oscillations in epidermal and guard cells in response to the fungal elicitor chitin. Moreover, we discovered that flg22- and chitin-induced Ca2+ signals in the root initiate from the elongation zone.
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影响因子：
56.9
作者：
IKURA, M;CLORE, GM;BAX, A
通讯作者：
BAX, A
DOI：
10.1083/jcb.144.3.483
发表时间：
1999-02-08
期刊：
The Journal of cell biology
影响因子：
--
作者：
Feijó JA;Sainhas J;Hackett GR;Kunkel JG;Hepler PK
通讯作者：
Hepler PK
影响因子：
4.8
作者：
Heim, N;Griesbeck, O
通讯作者：
Griesbeck, O
影响因子：
7.2
作者：
Grefen, Christopher;Donald, Naomi;Blatt, Michael R.
通讯作者：
Blatt, Michael R.
影响因子：
1.2
作者：
Grossmann, Guido;Meier, Matthias;Frommer, Wolf B.
通讯作者：
Frommer, Wolf B.