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Purpose: To investigate the effects of gamma pre-exposure on cadmium accumulation in Plantago ovata seedlings. Metallothionein (MT) localization was also studied following Cadmium (Cd) treatment in P. ovata. Materials and methods: DNA damage was determined by alkaline comet assay. MT gene and protein expression were studied by real-time polymerase chain reaction and flow cytometry, respectively, in root and shoot tissues. Metal accumulation (Cd, zinc [Zn], iron [Fe]) was evaluated by Atomic Absorption Spectroscopy. Results: Cd treatment decreased seed germination rate, biomass and free radical scavenging activity and increased DNA damage in a dose-dependent manner. When P. ovata seeds were pre- exposed to 5 Gy gamma dose (prior to Cd treatment) seed germination rate, biomass and free radical scavenging activity increased significantly. MT genes (PoMT1, PoMT2 and PoMT3) and MT protein expression enhanced when 5 Gy gamma-irradiated seeds were grown in Cd containing medium and Cd accumulation also increased in a dose-dependent manner. Conclusions: Higher Cd accumulation in P. ovata seedlings may be attributed to the upregulation of PoMT genes in gamma pretreated seedlings. Localization of metallothionein in cytosol and nucleus indicated its positive role against Cd-mediated cytotoxic and genotoxic effects. [ABSTRACT FROM AUTHOR]

Purpose: To study the oxidative stress-induced changes by sub-lethal doses of cadmium chloride (CdCl2) and gamma irradiation, two redox-inducing agents, on metallothionein (MT) gene and protein expression in Plantago ovata Forsk (P. ovata). Materials and methods: Chlorophyll content was estimated to study the stress response in P. ovata seedlings following exposure to gamma irradiation and CdCl2. Lipid peroxidation and proline content, two oxidative stress markers, were also studied. The level of metallothionein gene and protein expression was further investigated using polymerase chain reaction (PCR), reverse transcription-PCR (RT-PCR), immunocytochemistry and flow cytometry. Results: Three MT genes of P. ovata namely PoMT 1, PoMT2 and PoMT 3 were isolated, sequenced and characterized and their expressions were found to be altered in the case of both oxidative stresses in a dose-dependent and tissue-specific manner. The results were in agreement with the observations from immunocytochemistry and FACS analysis. Conclusions: The results suggest that both gamma irradiation and CdCl2 alter redox balance in P. ovata. The metallothionein gene may play an important role in metal tolerance and stress balance. It is conjectured that the stress-mediated imbalance is maintained by altered MT gene and protein expression. [ABSTRACT FROM AUTHOR]

Purpose: The effect of gamma rays on metallothionein (MT) expression was studied using the medicinal plant Plantago ovata as the test system. Materials and methods: Western blotting and Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) were used for this purpose. Results: Western blot analysis showed significant induction of metallothionein protein following gamma exposure and that induction was highest at 20 Gy gamma dose. At higher gamma doses (100 Gy) MT expression level declined due to degeneration of cells. MALDI-TOF MS analysis indicated the presence of low molecular weight (7-8 kD) MT molecules following the lower radiation doses. Conclusion: It was concluded from the MALDI-TOF MS result that low gamma exposure leads to expression of MT-like protein. At high doses of gamma ray, MT homologues or MT-like protein were not identified, possibly because they might have precipitated due to uncontrolled cross-linking and non-specific aggregation. [ABSTRACT FROM AUTHOR]