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Response of antioxidative defense system in two ecotypes of Arabidopsis thaliana (Col-0 & Ler-0) during mercury-induced oxidative stressLiu, Chien-Shin 28 July 2011 (has links)
Generation of reactive oxygen species (ROS) is an important view point to evaluate heavy metal toxicity and resistance in plants. Arabidopsis thaliana is a fully sequenced model plant, and the characteristic between ecotypes due to adaptation towards varied environment can be used as a material for comparing physiological differences. In this experiments, two ecotypes of A. thaliana: Columbia (Col-0) and Landsberg erecta (Ler-0) is observed for the roots growth inhibition, plasma membrane integrity and lipid peroxidation after treated with different concentration of HgCl2 (0, 2, 4, 8 £gM), in attempt to compare the anti-oxidation defensive mechanism of two ecotypes and understand mercury-induced oxidative stress. ROS and Ca2+ generation is determined with CM-H2DCF-DA and Oregon Green 488 BAPTA-1 is under confocal microscopy. Some anti-oxidant enzymes such as superoxide dismutase (SOD EC 1.15.1.1), peroxidase (POD EC1.11.1.7) and ascorbate peroxidase (APX EC 1.11.1.11) are examined for the activity under protein gel electrophoresis. Experiment results showed that mercury-induced inhibition of root growth is more significant in Ler-0. ROS in roots of both ecotypes shows different trends under 8 £gM HgCl2 , however increment of ROS level below 4 £gM HgCl2 ; Ca2+ shows the similar result as ROS. Activity of SOD isoforms reached a peak at 2-4 £gM HgCl2. Expression of POD is correlated to the mercury concentration in both ecotypes. There are two types of APXs expression, one decreased as mercury concentration increased, another increased under 2 £gM HgCl2 and decreased as the concentration getting higher. According to the observation on expression of ROS generations and anti-oxidation system, we speculated that tolerance of Ler-0 towards mercury is weaker than Col-0. The results can be used as a basis for further discussion on influence of mercury towards different anti-oxidation enzymes and the signaling pathways.
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