Uptake of aluminium into Arabidopsis root cells measured by fluorescent lifetime imaging

Authors: Babourina, Olga; Rengel, Zed

Source: Annals of Botany, Volume 104, Number 1, 15 July 2009 , pp. 189-195(7)

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Abstract:

Background and Aims

Measuring the Al³ uptake rate across the plasma membrane of intact root cells is crucial for understanding the mechanisms and time-course of Al toxicity in plants. However, a reliable method with the sufficient spatial and temporal resolution to estimate Al³ uptake in intact root cells does not exist.

Methods

In the current study, fluorescent lifetime imaging (FLIM) analysis was used to quantify Al³ uptake in the root-cell cytoplasm in vivo. This was performed via the estimation of the fluorescence lifetime of Allumogallion 5-chloro-3[(2,4-dihydroxyphenyl)azo]-2-hydroxybenzenesulfonic acid complexes and measurements of intracellular pH while exposing arabidopsis seedlings to acidic and Al³ stresses.

Key Results

The lifetime of Allumogallion complexes fluorescence is pH-dependent. The primary sites for Al³ entry are the meristem and distal elongation zones, while Al³ uptake via the cortex and epidermis of the mature root zone is limited. The maximum rates of Al uptake into the cytoplasm (23 mol m³ min¹ for the meristematic root zone and 37 mol m³ min¹ for the mature zone) were observed after a 30-min exposure to 100 m AlCl₃ (pH 42). Intracellular Al concentration increased to 04 m Al within the first 3 h of exposure to 100 m AlCl₃.

Conclusions

FLIM analysis of the fluorescence of Allumogallion complexes can be used to reliably quantify Al uptake in the cytoplasm of intact root cells at the initial stages of Al³ stress.

Keywords: Acid stress; Al3; aluminium toxicity; Arabidopsis thaliana; low pH; fluorescent lifetime imaging (FLIM); lumogallion

Document Type: Short communication

DOI: http://dx.doi.org/10.1093/aob/mcp098

Publication date: 2009-07-15

More about this publication?

Annals of Botany is an international plant science journal with editorial offices in Australia, China, Japan, Mainland Europe, UK and USA. It is published monthly in both electronic and printed forms with at least one extra issue each year that focuses on a particular theme in plant biology. The Journal is managed by the Annals of Botany Company, a not-for-profit educational charity established to promote plant science worldwide.

Uptake of aluminium into Arabidopsis root cells measured by fluorescent lifetime imaging

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