Imaging plant cell death: GFP-Nit1 aggregation marks an early step of wound and herbicide induced cell deathReportar como inadecuado




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BMC Plant Biology

, 5:4

First Online: 29 March 2005Received: 09 December 2004Accepted: 29 March 2005

Abstract

BackgroundA great deal is known about the morphological endpoints of plant cell death, but relatively little is known about its sequence of events and - or its execution at the biochemical level. Live cell imaging using GFP-tagged markers is a powerful way to provide dynamic portraits of a cellular process that can in turn provide a descriptive foundation valuable for future biochemical and genetic investigations.

ResultsWhile characterizing a collection of random GFP-protein fusion markers we discovered that mechanical wounding induces rapid aggregation of a GFP-Nitrilase 1 fusion protein in Arabidopsis cells directly abutting wound sites. Time-lapse imaging of this response shows that the aggregation occurs in cells that subsequently die 30 – 60 minutes post-wounding, indicating that GFP-Nit1 aggregation is an early marker of cell death at wound sites. Time-lapse confocal imaging was used to characterize wound-induced cell death using GFP-Nit1 and markers of the nucleus and endoplasmic reticulum. These analyses provide dynamic portraits of well-known death-associated responses such as nuclear contraction and cellular collapse and reveal novel features such as nuclear envelope separation, ER vesiculation and loss of nuclear-lumen contents. As a parallel system for imaging cell death, we developed a chemical method for rapidly triggering cell death using the herbicides bromoxynil or chloroxynil which cause rapid GFP-Nit1 aggregation, loss of nuclear contents and cellular collapse, but not nuclear contraction, separating this response from others during plant cell death.

ConclusionOur observations place aggregation of Nitrilase 1 as one of the earliest events associated with wound and herbicide-induced cell death and highlight several novel cellular events that occur as plant cells die. Our data create a detailed descriptive framework for future investigations of plant cell death and provide new tools for both its cellular and biochemical analysis.

Electronic supplementary materialThe online version of this article doi:10.1186-1471-2229-5-4 contains supplementary material, which is available to authorized users.

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Autor: Sean R Cutler - Chris R Somerville

Fuente: https://link.springer.com/







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