• Molecules and Cells
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• v.19 no.1
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• pp.81-87
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• 2005

Phytochelatins play an important role in heavy metal detoxification in plants as well as in other organisms. The Arabidopsis thaliana mutant cad1-3 does not produce detectable levels of phytochelatins in response to cadmium stress. The hypersensitivity of cad1-3 to cadmium stress is attributed to a mutation in the phytochelatin synthase 1 (AtPCS1) gene. However, A. thaliana also contains a functional phytochelatin synthase 2 (AtPCS2). In this study, we investigated why the cad1-3 mutant is hypersensitive to cadmium stress despite the presence of AtPCS2. Northern and Western blot analyses showed that expression of AtPCS2 is weak compared to AtPCS1 in both roots and shoots of transgenic Arabidopsis. The lower level of AtPCS2 expression was confirmed by RT-PCR analysis of wild type Arabidopsis. Moreover, no tissue-specific expression of AtPCS2 was observed. Even when AtPCS2 was under the control of the AtPCS1 promoter or of the cauliflower mosaic virus 35S promoter (CaMV 35S) it was not capable of fully complementing the cad1-3 mutant for cadmium resistance.

• Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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• 2004.09a
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• pp.290-292
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• 2004

Heavy metal contamination has been increased in aqueous environments near many industrial facilities, such as metal plating facilities, mining operations, and tanneries. The soils in the vicinity of many military bases are also reported to be contaminated and pose a risk of groundwater and surface water contamination with heavy metals. The biological removal of metals through bioaccumulation has distinct advantages over conventional methods; the process rarely produces undesirable or deleterious chemical byproducts, it is highly efficient, easy to operate and cost-effective in the treatment of large volumes of wastewater containing toxic heavy metals. In addition, a recent development of molecular biology shed light on the enhancing the microorganism's natural remediation capability as well as improving the current biological treatment. In this study, characteristics of the cell growth and heavy metal accumulation by Saccharomyces cerevisiae strains expressing phytochelatin syntahse (PCS) gene were studied in batch cultures. The AtCRFI gene was demonstrated to confer substantial increases in metal tolerance in yeast. PCS-expressing cells tolerated more Cd$^{2＋}$ than controls.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.14 no.6
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• pp.3114-3119
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• 2013

The mangrove ecosystems have the capacity to act as a sink of heavy metals entering aquatic ecosystems. Despite their potential exposure to metal contaminated sediments, mangroves appear to be highly tolerant to heavy metals. In this study, we cloned metal tolerance gene from mangrove plant. Using CTAB method, RNA were isolated from leaves and root tissue of Rhizophora stylosa habitated at Weno island in Micronesia Chuuk lagoon using CTAB method and phytochelatin synthase 1 (PCS1) gene was cloned using gene specific primers. Expression of PCS1 gene was increased 1.91 fold and 2.72 fold in mangrove propagules exposed to 100 ppb Cd and 10 ppb Cu, respectively. These results indicate that expression of PCS1 gene are promising tools for health assessment of mangrove ecosystem.

• Korean Journal of Environmental Agriculture
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• v.28 no.4
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• pp.409-411
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• 2009

Phytochelatins (PCs) are small-sized peptides synthesized by PC synthase (PCS) using glutathione (GSH) as a substrate, and they play an important role in the detoxification of toxic heavy metals in plants, fission yeast, and other living organisms. Recently, it has been suggested that PCS is also involved in degradation of some xenobiotics including monobromobimane. PCS cleaves the Gly residue from GSH-xenobiotics conjugates resulting in ${\gamma}$-Glu-Cys-xenobiotics, and this is to degraded further. Therefore, our research is focus on whether PCS is also involved in degradation of tolclofos-methyl, an important pesticide which has been used in ginseng cultivated areas. Heterologous expression of Arabidopsis PCS confers tolerance to tolclofos-methyl in yeast. Furthermore, PCS-deficient Cad1-3 Arabidopsis mutant showed high sensitivity to tolclofos-methyl compared with wild-type plants. These results imply that PCS is involved in degradation of tolclofos-methyl as other xenobiotics.

• Proceedings of the Zoological Society Korea Conference
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• 1995.10b
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• pp.263.1-263
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• 1995

No Abstract, See Full Text

• Applied Biological Chemistry
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• v.40 no.3
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• pp.243-248
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• 1997

In order to evaluate the usefulness of O. javanica for the phytoremediation, it was grown for 1, 3, 7, 14, 21 days and was exposed to $50\;{\mu}M\;of\;CdCl_2$ in hydroponic medium after 3 weeks. Its biomass and contents of chlorophylls were analyzed. The growth of O. javanica showed little difference between cadmium treated and non-treated groups, while its contents of chlorophylls of Cd-treated group decreased up to 50% compared to the case of non-treated group. Its accumulated cadmium concentrations were 2.1, 7.3 and $113\;{\mu}moles\;Cd/g$ dry weight in the leaf, stem and root, respectively. The total contents of thiol increased 0.5, 1 and 7 times in the leaf, stem and root, respectively, while the contents of glutathione tended to decrease by 43%, 70% and 47% in the leaf, stem and root, respectively. Using HPLC analysis, the reasonable peaks of thiol compounds in shoot and root of Cd-treated sample were compared to those of non-treated sample in O. javanica, and found to be phytochelatins. In case of Nicotiana tabacum cv. Xanthi tested as control plant, the cadmium treatment for 3 weeks resulted in the decrease of both biomass and chlorophyll up to 70% and 75%, respectively. The roots of tobacco became rotten and eventually died. These results suggested that Oenanthe javanica is cadmium-tolerant hyperaccumulator.(Received December 20, 1996; accepted March 17, 1997)

• The Korean Journal of Ecology
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• v.26 no.5
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• pp.263-266
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• 2003

This research investigated the process of removing cadmium and tested the detoxification mechanism of the cadmium-binding peptide (Cd-BP) from Rumex crispus. Phytochelatin-like cadmium-binding peptide (PC-Cd-BP) of Rumex crispus was purified and identified. Rumex crispus was exposed to 4.3 mg Cd/L for seven days. Heat-treated supernatant fraction taken by root tissues showed traces of PC-Cd-BP An analysis of the material through Gel-filteration chromatography on the Sephadex G-75 column showed two symmetrical Cd-BP peaks. The major peak with the smaller molecular weight was further purified by $C_{18}$ reverse-phase HPLC to produce apparent homogeneity. The amino acid composition of Cd-BP from Rumex crispus included cysteine (22.6%), glutamate and glutamate acid (20%), and glycine (12%). It was similar the amino acid composition of most PC. The molecular weight of the purified peptide was determined at 568-706 Da by MALDI-TOF MS. Therefore, the Cd-BP of Rumex crispus was PC-Cd-BP consisting of isopeptides.

• Korean Journal of Environmental Biology
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• v.18 no.2
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• pp.269-277
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• 2000

When Persicaria thunbergii and Rumex crispus were treated with Cd($NO_3$)$_2$ and Pb($NO_3$)$_2$ of 5 or 10 mM for 5 days, the amount of bioaccumulation of $Pb^{2+}$ in the leaf of P. thunbergii was 2.87-8.08$\mu\textrm{g}$/g and that of $Cd^{2+}$ was 0.82-2.79$\mu\textrm{g}$/g. In the case of P. thunbergii, the concentration of $Pb^{2+}$ in the leaf was higher than that of $Cd^{2+}$. On the other hand, in R. crispus, the concentration of $Cd^{2+}$ and $Pb^{2+}$ were similar as follows ; 1.49$\mu\textrm{g}$/g in $Cd^{2+}$ 5mM, 2.90$\mu\textrm{g}$/g in Cd2+ 10mM, 1.83$\mu\textrm{g}$/g in $Pb^{2+}$ 5mM and 2.73$\mu\textrm{g}$/g in $Pb^{2+}$ 10mM. The remaining rate of heavy metals and the variation of pH in the cultured soil decreased as compared with control (100 % and pH 6.48) after 5 days as follows; to 77.l% and pH 6.39 in $Cd^{2+}$ 5mM, 90.2% and pH 5.79 in $Cd^{2+}$ 10 mM, 81.1% and pH 6.00 in $Pb^{2+}$ 5mM, and 85.7% and pH 5.80 in $Pb^{2+}$ 10 mM. The result of size exclusion chromatography, several phytochelatins were seperated from the extract of the leaf of both plants treated with heavy metals. The molecular mass of these phytochelatins were estimated as follows; in the case of P. thunbergii, about 4,300-8,600 da by $Cd^{2+}$ and about 3,200-9,700 da by $Pb^{2+}$, and in R. crispus, about 4,300 da by $Cd^{2+}$ and about 3,200-7,500 da by $Pb^{2+}$. In addition, $A_{254}$ of these phytochelatins were higher than $A_{280}$. [Phytochelatin, Persicaria thunbergii, Rumex crispus]

• Journal of Korea Soil Environment Society
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• v.5 no.1
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• pp.33-43
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• 2000

For the consideration of phytoremediation, TEX>$Cd^{2+}$ and Pb$^{2+}$ were analysed in the soil of the habitats and the leaf stem and root of Persicaria thunbergii in the different localities of Bong-Dong river In the soil and plant samples of research areas, TEX>$Cd^{2+}$ was not detected but, $Pb^{2+}$ detected as follows; about 7.5~15.5$mu\textrm{g}$/g in the soil of habitats, about 11.7~18.4 $mu\textrm{g}$/g in the leaf, about 7.~15.5$mu\textrm{g}$/g in the stem and about 89.1~193.6$\mu\textrm{g}$/g in the root of P. thunebrgii and the correlation coefficient value between the $Pb^{2+}$ contents in soil and P. thunbergii was 0.814(>t12, 0.01). After P thunbergii was treated with Cd(NO$_3$)$_2$and Pb(NO$_3$)$_2$of 5 and 10mM, the bioaccumulation of TEX>$Cd^{2+}$ and $Pb^{2+}$ in the leaf of plant, the remaining mass of heavy metals and the variation of pH in the soil, and the increasing rate(%) of phytochelatin in plant were examined. The concentrations of TEX>$Cd^{2+}$and $Pb^{2+}$ in the leaf as follows, in the case of TEX>$Cd^{2+}$ about 0.82~2.79$\mu\textrm{g}$/g and in $Pb^{2+}$, about 2.87~8.08$\mu\textrm{g}$/g. The remaining mass of heavy metals and the variation of pH in the cultured soil decreased as follows; about 77.1% and pH6.39 in TEX>$Cd^{2+}$ 5mM, about 90.2% and pH5.79 in TEX>$Cd^{2+}$ 10mM, about 81.1% and pH6.00 in $Pb^{2+}$ 5mM and about 85.7% and pH5.80 in Pb$^{2+}$ 10mM. The phytochelatin were increased in plant samples treated with 10mM Cd(NO$_3$)$_2$and Pb(NO$_3$)$_2$as follows; about 259% by TEX>$Cd^{2+}$ and about 305% by Pb$^{2+}$ be compared with control. and the molecular weight(da) of these phytochelatins were estimated about 4,300~8,600da in the case of the treatment of TEX>$Cd^{2+}$ and about 3,200~9,700 in $Pb^{2+}$./TEX>.

• Proceedings of the Botanical Society of Korea Conference
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• 2002.04a
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• pp.62-72
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• 2002

This study has investigated the biosynthesis and function of the heavy metal binding peptides, the phytochelatins, in plants. PCs are synthesised enzymatically from glutathione by the enzyme PC synthase in the presence of heavy metal ions. Using Arabidopsis thaliana as a model organism cadmium-sensitive, phytochelatin-deficient mutants have been isolated and characterised in previous studies. The cadl mutants have wildtype levels of glutathione, are PC deficient and lack PC synthase activity. Thus, the CADl gene has been proposed to encode PC synthase. The CADl gene was isolated by a positional cloning strategy The gene was mapped and a candidate identified. Each of four cadl mutants had a single base pair change in the candidate gene and the cadmium-sensitive, cadl phenotype was complemented by the candidate gene. This demonstrated the CADl gene had been cloned. A homologous gene in the fission yeast, Schizosaccharomyces pombe was identified through database searches. A targeted-deletion mutation of this gene was constructed and the mutant, like cadl mutants of Arabidopsis, was cadmium-sensitive and PC-deficient. A comparison of the redicted amino acid sequences reveals a highly conserved N-terminal region Presumed to be the catalytic domain and a variable C-terminal region containing multiple Cys residues proposed to be involved in activation of the enzyme by metal ions. Similar genes were also identified in animal species. The Arabidopsis CADl/AtPCSl and S. pombe SpbPCS genes were expressed in E. coli and were shown to be sufficient for glutathione-dependent, heavy metal activate PC synthesis in vitro, thus demonstrating these genes encode PC synthase enzymes. Using RT-PCR, AtPCSl expression appeared to be independent of Cd exposure. However, at higher levels of Cd exposure a AtPCSl-CUS reporter gene construct appeared to be more highly expressed. Using the reporter gene construct, AtPCSl was expressed most tissues. Expression appeared to be greater in younger tissues and same higher levels of expression was observed in some regions, including carpels and the base of siliques. AtPCS2 was a functional gene encoding an active PC synthase. However, its Pattern of expression and the phenotype of a mutant (or antisense line) have not been determined. Assuming the gene is functional then it has clearly been maintained through evolution and must provide some selective advantage. This implies that, at least in some cells or tissue, it is likely to be the dominant PC synthase expressed. This remains to be determined