Effect of nitric oxide on concentration of intracellular free Ca2+ in transgenic Arabidopsis thalianaplants during oxidative stress
© The Author(s) 2005
Published: 31 May 2005
In recent years exogenous NO was shown to (i) influence plant growth and development, (ii) take part in the response of plants to pathogens, light and ABA-induced stomatal closure, and (iii) reduce consequences of oxidative stress generated by treatment with herbicides [1–4]. It is also known that a number of chemical and physical stimuli, including oxidative stress, mediates their effects via transient increases in the concentration of intracellular free Ca2+([Ca2+]cyt). Therefore, the aim of our work was to study the influence of exogenous NO on the increase of [Ca2+]cyt induced by oxidative stress in plant cells with changes in [Ca2+]cyt during oxidative stress being measured in Arabidopsis thaliana seedlings transformed to express apoaequorin.
Materials and methods
8-day old seedlings of A. thaliana were incubated in coelenterazine (5 μM) diluted in methanol for 6 h to reconstitute the aequorin. Chemiluminescence measurements were performed with a digital chemiluminometer. The peak-value of the stimuli-induced [Ca2+]cyt transient was calculated as a described by Cobbold and Rink  with some modifications . Oxidative stress induced by hydrogen peroxide (10 mM) . To evaluate a possible effect of NO on the increase [Ca2+]cyt during the oxidative stress seedlings were pre-treated with NO donor NOR-1 (25 × 10-6 M).
These data suggest that NO plays an important role in the activation of plant defense responses after oxidative stress. It may partly explain that NO is able to inactivate directly the reactive oxygen species (ROS) . The presence of an unpaired electrons within the NO molecule gives it its reactive species properties and is also the origin of its duality. At physiological concentrations NO may play a protective role acting as a chain inhibitor to limit the damage.
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