Phytochelatin Synthase Gene Research Articles

Homologous cloning, characterization and expression of a new halophyte phytochelatin synthase gene in Suaeda salsa

The halophyte Suaeda salsa can grow in heavy metal-polluted areas along intertidal zones having high salinity. Since phytochelatins can eff ectively chelate heavy metals, it was hypothesized that S. salsa possessed a phytochelatin synthase (PCS) gene. In the present study, the cDNA of PCS was obtained from S. salsa (designated as SsPCS) using homologous cloning and the rapid amplification of cDNA ends (RACE). A sequence analysis revealed that SsPCS consisted of 1 916 bp nucleotides, encoding a polypeptide of 492 amino acids with one phytochelatin domain and one phytochelatin C domain. A similarity analysis suggested that SsPCS shared up to a 58.6% identity with other PCS proteins and clustered with PCS proteins from eudicots. There was a new kind of metal ion sensor motif in its C-terminal domain. The SsPCS transcript was more highly expressed in elongated and fibered roots and stems (P<0.05) than in leaves. Lead and mercury exposure significantly enhanced the mRNA expression of SsPCS (P<0.05). To the best of our knowledge, SsPCS is the second PCS gene cloned from a halophyte, and it might contain a diff erent metal sensing capability than the first PCS from Thellungiella halophila. This study provided a new view of halophyte PCS genes in heavy metal tolerance.

Influence of Sulfur on Transcription of Genes Involved in Arsenic Accumulation in Rice Grains

Arsenic (As) contamination of rice grains affects millions of people worldwide. In this study, we found that sulfur application (20As+120S) decreased As concentration in rice grains by 44 % compared to grains without sulfur application (20As+0S). Importantly, sulfur application decreased arsenate [As(V)] and arsenite [As(III)] concentration in rice grains significantly, while there was no significant effect on dimethylarsenate (DMA) concentration. To elucidate the molecular basis of As accumulation in rice grains, we performed Illumina sequencing to acquire the differentially expressed genes induced by arsenate and sulfur treatments. By contrast with the control, the expression of 1,000 genes was found to be changed significantly, with 46 genes up-regulated and 954 genes down-regulated in grains grown in arsenate-contaminated soil (20As+0S). Between samples of control and arsenate together with sulfur treatment (20As+120S), 1,169 genes expressed significantly differently, with 16 genes up-regulated and 1,153 genes down-regulated. Sulfur application significantly changed the expression of genes involved in As metabolism in rice grains, significantly down-regulated phosphate transporter gene OsPT23 and aquaporin gene OsTIP4;2, while ABC transporter genes (OsABCG5, OsABCI7_2 and OsABC6) and phytochelatin synthase genes (OsPCS1, OsPCS3 and OsPCS13) were up-regulated. These results provide an insight into the molecular basis of how sulfur assimilation regulates As accumulation in rice grains.

Dermal exposure of Eisenia andrei earthworms: Effects of heavy metals on metallothionein and phytochelatin synthase gene expressions in coelomocytes.

Parameters such as total number of coelomocytes, riboflavin content in coelomocytes, expression of genes implied in metal homeostasis, and detoxification mechanisms can be used as biomarkers to assess the impact of metals on annelids. Defense biomarkers (detoxification gene expressions and coelomocyte parameters) were investigated in the ecotoxicologically important species Eisenia andrei following in vivo exposure to 5 different metals (zinc, copper, nickel, lead, and cadmium) at known concentrations. Coelomocyte numbers and riboflavin content were not affected by metallic exposure, but metal-specific gene expression variations were evidenced.

Overexpression of PtPCS enhances cadmium tolerance and cadmium accumulation in tobacco

Phytochelatins chelate heavy metal ions to decrease their toxicity. The chelates are then transferred to, and stored in, the vacuole. Phytochelatin synthase (PCS), which is involved in phytochelatin synthesis, is thought to be a key enzyme for phytoremediation. In this study, a PCS gene encoding phytochelatin synthase was cloned from poplar (Populus tomentosa Carr.), a widely grown model woody plant that accumulates high levels of heavy metals, especially cadmium. Poplar is considered to have potential applications in phytoremediation. The full-length PtPCS cDNA (1512-bp) encoded a polypeptide of 503 amino acid residues. The PtPCS cDNA was transferred into tobacco by Agrobacterium-mediated leaf disk transformation. The transgenic and wild-type (WT) lines of tobacco were subjected to a one time Cd treatment (90 μmol Cd2+) for 30 days, and then evaluated to determine their Cd tolerance. We evaluated morphological and physiological indices including leaf relative electrolyte leakage, malondialdehyde content, total superoxide dismutase activity, chlorophyll content and root activity. Compared with WT plants, the transgenic plants expressing PtPCS grew better in the Cd treatment and showed significantly higher Cd tolerance. Compared with WT plants, the transgenic lines accumulated higher concentrations of Cd (1.7 to 3.0-fold higher Cd concentration in roots; 1.24 to 2.28-fold higher Cd concentration in leaves). However, the transfer coefficient was lower in the transgenic lines than in wild type. We concluded that PtPCS encodes a functional PCS that may be involved in Cd tolerance and accumulation, but not in Cd transport.

The construction of an engineered bacterium to remove cadmium from wastewater.

The removal of cadmium (Cd) from wastewater before it is released from factories is important for protecting human health. Although some researchers have developed engineered bacteria, the resistance of these engineered bacteria to Cd have not been improved. In this study, two key genes involved in glutathione synthesis (gshA and gshB), a serine acetyltransferase gene (cysE), a Thlaspi caerulescens phytochelatin synthase gene (TcPCS1), and a heavy metal ATPase gene (TcHMA3) were transformed into Escherichia coli BL21. The resistance of the engineered bacterium to Cd was significantly greater than that of the initial bacterium and the Cd accumulation in the engineered bacterium was much higher than in the initial bacterium. In addition, the Cd resistance of the bacteria harboring gshB, gshA, cysE, and TcPCS1 was higher than that of the bacteria harboring gshA, cysE, and TcPCS1. This finding demonstrated that gshB played an important role in glutathione synthesis and that the reaction catalyzed by glutathione synthase was the limiting step for producing phytochelatins. Furthermore, TcPCS1 had a greater specificity and a higher capacity for removing Cd than SpPCS1, and TcHMA3 not only played a role in T. caerulescens but also functioned in E. coli.

Comprendiendo los mecanismos de hiperacumulación de Cd en S. kali, una planta del desierto

Salsola kali, una planta desértica, ha sido propuesta como un potencial hiperacumulador de Cd. Estudios de espectroscopía de Rayos-X han mostrado que compuestos conteniendo oxígeno y grupos tiol están involucrados en la absorción de Cd en esta planta. Por ello se ha planteado que algunos ácidos orgánicos, pared celular, fitoquelatinas y otros compuestos conteniendo glutatión, pueden estar relacionados con los mecanismos de hiperacumulación de Cd en S. kali. En esta investigación se expusieron plantas silvestres a diferentes concentraciones de Cd para determinar el contenido de este metal en el xilema y floema, así como en fracciones proteínicas. Adicionalmente, extractos de las plantas se analizaron mediante cromatografía líquida a alta presión para identificar y cuantificar ácidos orgánicos. Las plantas se trataron con 0, 20, 200 y 400 mg Cd L-1 por 48 h en hidroponia. La incorporación de Cd se midió por separado en raíces, floema y xilema. Se encontró que en plantas tratadas con concentraciones de Cd por encima de los 200 mg Cd L-1, el contenido de Cd fue mayor en el floema que en el xilema. El perfil proteínico (SDS-PAGE) mostró que en plantas tratadas con Cd se incrementa la presencia de dos péptidos y se expresa uno nuevo. Después de una filtración en gel G25 y codeterminación de Cd, se encontró que es muy probable que dos proteínas (de 29 kDa y 14 kDa) estén asociadas al Cd. El uso de primers degenerados de la familia Brassica permitió la identificación de un posible gen de la fitoquelatin sintasa. Los ácidos cítrico y oxálico fueron identificados en los extractos de las plantas. No se encontraron diferencias significativas entre las concentraciones de ácido cítrico en plantas control y en aquellas tratadas con Cd. Por otro lado, la cantidad de ácido oxálico en plantas expuestas a Cd fue significativamente menor que en las plantas control. Estos datos pueden indicar que el Cd pudo haber precipitado como cristales de oxalato. Los resultados reportados aquí serán útiles para entender a mayor profundidad los mecanismos de hiperacumulación de Cd en S. kali.

Transcriptional and physiological changes in relation to Fe uptake under conditions of Fe-deficiency and Cd-toxicity in roots of Vigna radiata L.

We investigated transcriptional and physiological changes in relation to Fe transport and uptake under various conditions of iron (Fe)-deficiency and cadmium (Cd) toxicity. Responses to four such Fe/Cd conditions were evaluated, revealing that oxidative stress was generated in the presence of Cd, followed by a decrease in Fe and an increase in Cd concentrations in green gram (Vigna radiata) material, whereas supplementation with Fe had a protective effect against Cd toxicity. The involvement of enzymes in Fe-uptake for the formation of root-nodules was largely reduced in the presence of Cd toxicity, a condition recovered by Fe-supplementation. Insufficient ferric chelate reducing activity in Fe-deprived roots in the presence of Cd was also largely improved by Fe supplementation. The expression of Fe(2+) transporters (IRT1, IRT2, and IRT3), Fe(III) chelate reductase (FRO1-FRO8) and phytochelatin synthase (PCS1, PCS2 and PCS3) genes was up regulated for the first 5 days and decreased after 10 days in roots in the presence of Cd toxicity, but was sustained with Fe-supplementation. Additionally, root biomass was fully recovered in plants in the presence of Fe during Cd toxicity. Our results suggest that Fe-status plays a significant role in ameliorating the damage in Fe transport for chelation and its uptake caused by Cd toxicity. This supports the hypothesis that leguminous plants, particularly those that are sensitive to Fe such as green gram, can cope to some extent with Cd toxicity by improving the uptake and transport of Fe.

Arbuscular mycorrhizal fungi play a role in protecting roots of Sophora viciifolia Hance. from Pb damage associated with increased phytochelatin synthase gene expression

Understanding the influence of arbuscular mycorrhizal (AM) fungi on the expressions of the dominant plant-related genes under heavy metal (HM) stress is important for developing strategies to reclaim polluted sites. In this study, we cloned full-length cDNAs of phytochelatin synthase gene (PCS1) and Actin of Sophora viciifolia Hance., a predominant plant in Qiandongshan lead and zinc mine, by rapid amplification of cDNA ends. Consequently, we studied the response of SvPCS1 to Funneliformis mosseae inoculation under lead stress (0, 50, and 200 μM Pb(NO3)2) at different durations (1, 3, and 7 days) using quantitative reverse-transcription polymerase chain-reaction (qRT-PCR) technique. The Pb concentrations and chlorophyll fluorescence parameters were also measured to assay Pb toxicity to Sophora viciifolia. We found that Pb concentrations in roots increased with increasing Pb application and the durations; the F v /F m , F v /F o , qP, and Y(II) decreased; NPQ rose with increasing Pb concentrations; mycorrhizal symbiosis alleviated the Pb toxicity to plants; and SvPCS1 was constitutively expressed in the roots. It was also found that F. mosseae inoculation could promote the expression of SvPCS1 with the concentration ≤ 200 μM at the exposure time shorter than 7 days.

Artificial induction of cadmium tolerance and its further enhancement via heterologous co-expression of SpHMT1 and AtPCS1 in the yeast cells

Microorganisms can be used in the bioremediation of heavy metals. It is desirable if the microorganisms show a strong tolerance as well as the ability to accumulate (or biosorb) heavy metals. Yeast is well known for having a high capacity of biosorption of heavy metals. Therefore, the present study focused on developing a yeast mutant that has a strong tolerance to cadmium (Cd), a representative toxic metal. The Cd-resistant yeast mutant (CdR) was induced and isolated by growing yeast cells in media containing Cd and gradually increasing the concentration until reaching a possible maximum of 20 mM CdCl2, to which the cells adapted and survived. CdR cells showed stronger tolerance to Cd-induced stress than the control cells. To obtain even higher Cd tolerance in the yeast cells, both Schizosaccharomyces pombe heavy metal tolerance factor 1 (SpHMT1) and Arabidopsis thaliana phytochelatin synthase (AtPCS1) genes were introduced into the CdR cells to be expressed simultaneously. The transformed CdR cells showed higher Cd tolerance than the untransformed CdR cells.

Characterization and transcription studies of a phytochelatin synthase gene from the solitary tunicate Ciona intestinalis exposed to cadmium

The major thiol-containing molecules involved in controlling the level of intracellular ROS in eukaryotes, acting as a nonenzymatic detoxification system, are metallothioneins (MTs), glutathione (GSH) and phytochelatins (PCs). Both MTs and GSH are well-known in the animal kingdom. PC was considered a prerogative of the plant kingdom but, in 2001, a phytochelatin synthase (PCS) gene was described in the nematode Caenorhabditis elegans; additional genes encoding this enzyme were later described in the earthworm Eisenia fetida and in the parasitic nematode Schistosoma mansoni but scanty data are available, up to now, for Deuterostomes. Here, we describe the molecular characteristics and transcription pattern, in the presence of Cd, of a PCS gene from the invertebrate chordate Ciona intestinalis, a ubiquitous solitary tunicate and demonstrate the presence of PCs in tissue extracts. We also studied mRNA localization by in situ hybridization. In addition, we analyzed the behavior of hemocytes and tunic cells consequent to Cd exposure as well as the transcription pattern of the Ciona orthologous for proliferating cell nuclear antigen (PCNA), usually considered a proliferation marker, and observed that cell proliferation occurs after 96h of Cd treatment. This matches the hypothesis of Cd-induced cell proliferation, as already suggested by previous data on the expression of a metallothionein gene in the same animal.

Expression of sulfur uptake assimilation-related genes in response to cadmium, bensulfuron-methyl and their co-contamination in rice roots

The responses of sulfur (S) uptake assimilation-related genes' expression in roots of two rice cultivars to cadmium (Cd), bensulfuron-methyl (BSM) and their co-contamination (Cd+BSM) were investigated by gene-chip microarray analysis and quantitative real-time PCR (QRT-PCR) technology. Treatments of Cd and Cd+BSM induced expression of sulfate transporter and permease genes, and promoted sulfate uptake in rice roots. Cd+BSM could alleviate Cd toxicity to cv. Fengmeizhan seedlings, probably due to Cd+BSM promoting greater S absorption by seedlings. Cd and Cd+BSM induced expression of sulfate assimilation-related genes, and thus activated the sulfur assimilation pathway. Cd and Cd+BSM induced expression of phytochelatin synthase and metallothionein genes, and induced expression of glutathione S-transferases (GSTs), glutathione synthase (GS) and S-containing antioxidation enzyme genes, which detoxified Cd2+. It is suggested that (to cope with the toxicity of Cd, BSM and their co-contamination) the S uptake and assimilation pathway was activated in rice roots by increased expression of related genes, thus enhancing the supply of organic S for synthesis of Cd or BSM resistance-related substances.

Metabolites and metals in Metazoa--what role do phytochelatins play in animals?

Phytochelatins are sulfur-rich metal-binding peptides, and phytochelatin synthesis is one of the key mechanisms by which plants protect themselves against toxic soft metal ions such as cadmium. It has been known for a while now that some invertebrates also possess functional phytochelatin synthase (PCS) enzymes, and that at least one species, the nematode Caenorhabditis elegans, produces phytochelatins to help detoxify cadmium, and probably also other metal and metalloid ions including arsenic, zinc, selenium, silver, and copper. Here, we review recent studies on the occurrence, utilization, and regulation of phytochelatin synthesis in invertebrates. The phytochelatin synthase gene has a wide phylogenetic distribution, and can be found in species that cover almost all of the animal tree of life. The evidence to date, though, suggests that the occurrence is patchy, and even though some members of particular taxonomic groups may contain PCS genes, there are also many species without these genes. For animal species that do possess PCS genes, some of them (e.g. earthworms) do synthesize phytochelatins in response to potentially toxic elements, whereas others (e.g. Schistosoma mansoni, a parasitic helminth) do not appear to do so. Just how (and if) phytochelatins in invertebrates complement the function of metallothioneins remains to be elucidated, and the temporal, spatial, and metal specificity of the two systems is still unknown.

Regulation of the Phytochelatin Synthase Gene &amp;lt;italic&amp;gt;AtPCS2 &amp;lt;/italic&amp;gt;in &amp;lt;italic&amp;gt;Arabidopsis thaliana&amp;lt;/italic&amp;gt;

Phytochelatin synthase (PCS) plays an essential role in heavy metal detoxification in plants and fungi, via the catalyzation of phytochelatins (PCs) biosynthesis from the substrate of glutathione (GSH). Two PCS genes <italic>AtPCS1</italic> and <italic>AtPCS2</italic> are present in <italic>Arabidopsis</italic> genome. However, loss of function of only the <italic>AtPCS1</italic> gene resulted in undetectable PCs and the consequent metal hypersensitivity in the corresponding mutant <italic>cad1</italic>-<italic>3</italic>,while <italic>in vitro</italic> assay indicated that AtPCS2 is capable to efficiently catalyze PCs biosynthesis. These results suggest that repression mechanisms exist in <italic>Arabidopsis</italic> to tightly control <italic>AtPCS2</italic> function <italic>in vivo</italic>. To test and decipher this hypothesis, we generated the construct 35S/AtPCS2::cMyc and transformed it into the PCs-deficient mutant <italic>cad1</italic>-<italic>3</italic>. Our results showed that both the <italic>AtPCS2</italic> mRNA and protein levels were steadily high in the transgenic plants. Consistently, PCs biosynthesis and metal tolerance were also observed when <italic>AtPCS2</italic> was introduced into <italic>cad1</italic>-<italic>3</italic>. Furthermore, in addition to the cytosol, AtPCS2 was also localized to the nuclei. Taken together, all these results indicate that <italic>AtPCS2</italic> is under tight control mainly at the transcription level in <italic>Arabidopsis</italic>, and it may function beyond catalyzing PCs biosynthesis.

RETRACTED ARTICLE: Coordinated response of sulfate transport, cysteine biosynthesis, and glutathione-mediated antioxidant defense in lentil (Lens culinaris Medik.) genotypes exposed to arsenic

Response of sulfate transporters, thiol metabolism, and antioxidant defense system was studied in roots of two lentil (Lens culinaris Medik.) genotypes grown in arsenic (10, 25, and 40 μM As(V))-supplemented nutrient solution, and significant changes compared to control (0 μM As(V)) were observed mainly at 25 and 40 μM. In L 414, high glutathione (GSH) redox (0.8-0.9) was maintained with elevated thiol synthesis, powered by transcriptional up-regulation of LcSultr1;1 and LcSultr1;2 sulfate transporters and significant induction of LcSAT1;1 and LcSAT1;2 (serine acetyltransferase), OAS-TL (O-acetylserine(thiol)-lyase), γ-ECS (γ-glutamylcysteine synthetase), and PCS (phytochelatin synthase) genes predominantly within 12-24 h of As exposure at 25 μM and within 6-12 h at 40 μM. This thiolic potency in L 414 roots was effectively complemented by up-regulation of gene expressions and consequent enhanced activities of superoxide dismutase, ascorbate peroxidase (APX), dehydroascorbate reductase, glutathione reductase (GR), and glutathione-S-transferase (GST) isoforms at 25 and 40 μMAs, efficiently scavenging excess reactive oxygen species to prevent onset of As-induced oxidative stress and consequent inhibition of root growth in L 414. In contrast, down-regulation of vital sulfate-uptake transporters as well as entire thiol-metabolizing system and considerably low APX, GST, and GR expressions in DPL 59 not only resulted in reduced GSH redox but also led to over-accumulation of H2O2. This triggered membrane lipid peroxidations as the marks of As-induced oxidative damage. Results indicated coordinated response of thiol-metabolism and antioxidant defense in conferring As-tolerance in lentil, and GSH is the key point in this cascade.

Earthworms Produce phytochelatins in Response to Arsenic

Phytochelatins are small cysteine-rich non-ribosomal peptides that chelate soft metal and metalloid ions, such as cadmium and arsenic. They are widely produced by plants and microbes; phytochelatin synthase genes are also present in animal species from several different phyla, but there is still little known about whether these genes are functional in animals, and if so, whether they are metal-responsive. We analysed phytochelatin production by direct chemical analysis in Lumbricus rubellus earthworms exposed to arsenic for a 28 day period, and found that arsenic clearly induced phytochelatin production in a dose-dependent manner. It was necessary to measure the phytochelatin metabolite concentrations directly, as there was no upregulation of phytochelatin synthase gene expression after 28 days: phytochelatin synthesis appears not to be transcriptionally regulated in animals. A further untargetted metabolomic analysis also found changes in metabolites associated with the transsulfuration pathway, which channels sulfur flux from methionine for phytochelatin synthesis. There was no evidence of biological transformation of arsenic (e.g. into methylated species) as a result of laboratory arsenic exposure. Finally, we compared wild populations of earthworms sampled from the field, and found that both arsenic-contaminated and cadmium-contaminated mine site worms had elevated phytochelatin concentrations.

Enhanced cadmium resistance and accumulation inPseudomonas putidaKT2440 expressing the phytochelatin synthase gene ofSchizosaccharomyces pombe

Phytochelatins (PCs) are cysteine-rich peptides with high binding affinity for toxic metals. Expressing the PC synthase gene (PCS) in plant growth-promoting bacteria may enhance its metal resistance and accumulation, consequently increasing phytoremediation efficiency in heavy metal pollution. In this study, PCS from Schizosaccharomyces pombe was cloned and expressed in Pseudomonas putida KT2440, which was confirmed by real-time RT-PCR through an increase in SpPCS mRNA expression level when induced by 20μmol of CdCl2 in the transformed Ps.putida cells. The recombined strain KT2440-SpPCS exhibited enhanced Cd, Ag and Hg resistance. Compared with the original strain, KT2440-SpPCS also displayed a threefold to fivefold increase in Cd accumulation (14·32μmol g(-1) to 17·38μmol g(-1) ; dry weight) when grown in 30 and 50μmol CdCl2 , along with an increase in nonprotein thiols. Further experiments showed significantly enhanced germination rates and growth of wheat seeds in 0·1mmol to 1·0mmol Cd when inoculated with KT2440-SpPCS. This study shows potential use of Ps.putida KT2440-SpPCS in plants to construct a symbiotic system for an enhanced phytoremediation of heavy metal-contaminated environments. The symbiotic system of using plant growth-promoting bacteria Pseudomonas putida to express phytochelatin synthase gene of Schizosaccharomyces pombe together in plants resulted in high heavy metal resistance and high accumulation capacity, suggesting potential enhancement in phytoremediation of heavy metal-contaminated environments.

Effect of cadmium on promoter activity of rice phytochelatin synthase gene in transgenic tobacco plants

The effect of cadmium acetate (200 and 400 μM Cd(CH3COO)2) on the transcriptional promoter activity of rice (Oryza sativa L.) phytochelatin synthase gene 900 bp in size was studied. The reporter gene of glucuronidase was placed under the control of this promoter in transgenic tobacco (Nicotiana tabacum L.) plants. For determining boundaries of this gene promoter responsive to cadmium, deletion constructs were created and their transcriptional activity was measured under the influence of different cadmium concentrations. Deletion of 189 bp from the 5′-terminus of this gene promoter resulted in some transcription activation, whereas deletion of successive 95 bp from the 5′-terminus resulted in cadmium-independent activation of the promoter.

Exposure of Brassica juncea (L) to arsenic species in hydroponic medium: comparative analysis in accumulation and biochemical and transcriptional alterations

Arsenic (As) contamination in the environment has attracted considerable attention worldwide. The objective of the present study was to see the comparative effect of As species As(III) and As(V) on accumulation, biochemical responses, and gene expression analysis in Brassica juncea var. Pusa Jaganath (PJn). Hydroponically grown 14-day-old seedlings of B. juncea were treated with different concentrations of As(III) and As(V). Accumulation of total As increased with increasing concentration of both As species and exposure time, mainly in roots. Reduction in seed germination, root-shoot length, chlorophyll, and protein content were observed with increasing concentration and exposure time of both As species, being more in As(III)-treated leaves. PJn variety showed that antioxidant enzymes (superoxide dismutase (SOD), catalase (CAT), and ascorbate peroxidase (APX)) and stress-related parameters (cysteine, proline, and malondialdehyde (MDA)) were stimulated and allows plant to tolerate both As species. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis in leaves showed significant changes in protein profile with more stringent effect with As(III) stress. Semiquantitative RT-PCR analysis showed regulation in expression of phytochelatin synthase (PCS), metallothionine-2 (MT-2), glutathione reductase (GR), and glutathione synthetase (GS) genes under both As(III) and As(V) stresses. Results suggested that accumulation and inhibition on physiological parameters differ according to the As species, while molecular and biochemical parameters showed a combinatorial type of tolerance mechanism against As(III) and As(V) stresses.

Expression of Ceratophyllum demersum phytochelatin synthase, CdPCS1, in Escherichia coli and Arabidopsis enhances heavy metal(loid)s accumulation

Phytochelatin synthase (PCS) gene encoding key enzyme for heavy metal detoxification and accumulation has been characterised from different sources and used to develop a technology for bioremediation. Past efforts provided limited success and contradictory results. Therefore, functional characterisation of PCS gene from new sources into different target systems is considered as an important task in the area of bioremediation. Earlier, we isolated and functionally characterised PCS gene from an aquatic macrophyte Ceratophyllum demersum L., a metal accumulator aquatic plant. Expression of this gene, CdPCS1, in tobacco enhanced PC synthesis and metal accumulation of transgenic tobacco plants. In the present study, we have expressed CdPCS1 in more diverse systems, Escherichia coli and Arabidopsis, and studied growth and metal accumulation of transgenic organisms. The expression of CdPCS1 in E. coli offered tolerance against cadmium as well as higher accumulation accompanied with PCS1 activity. The expression of CdPCS1 in Arabidopsis showed a significant enhanced accumulation of heavy metal(loid)s in aerial parts without significant difference in growth parameters in comparison to wild-type Arabidopsis plants. Our study suggests that CdPCS1 can be utilised for enhancing bioremediation potential of different organisms using biotechnological approaches.

Heteroexpression of the wheat phytochelatin synthase gene (&amp;lt;italic&amp;gt;TaPCS1&amp;lt;/italic&amp;gt;) in rice enhances cadmium sensitivity

Phytochelatin synthase (PCS )(EC 2.3.2.15 ) 催化 phytochelatins (PC ) 的最后的步生合成。PC 是在植物在重金属的隐遁和 detoxification 起一个重要作用的充满半胱氨酸的 thiol 反应、重的金属绑定肽的一个家庭。以前的研究显示了那种显示的那 overexpressed PCS 对比显型，从提高的镉(Cd )忍耐到在 Arabidopsis thaliana 的 Cd 超敏性。在这研究，小麦 phytochelatin synthase 基因， TaPCS1，是在野类型的米饭的 heteroexpressed (Oryza sativa L. ， cv。Zhonghua 11 ) 在米饭评估在 PC 和 Cd 忍耐的合成之间的关系。数据证明在米饭的 TaPCS1 的 heteroexpression 提高了 Cd 敏感并且显著地在射击，然而并非在根增加了 Cd 累积。另外， PCS 线在射击展出了 PC 和非蛋白质 thiols (NPT ) 的一个高得多的内容。在导致的 NPT 作文的突出的变化在射击在房间细胞器减少了谷胱甘肽水池弄空和更高的 Cd 内容，由更高氧化的应力列在后面，它可能导致 Cd 敏感。因此，在米饭的 TaPCS1 的 heteroexpression 能够在米饭射击增加 Cd 累积并且提高 Cd 敏感。