Search

Characterization of the typical petroleum pollutants, polycyclic aromatic hydrocarbons (PAHs) and n-alkanes, and indigenous microbial community structure and function in historically contaminated soil at petrol stations is critical. Five soil samples were collected from a petrol station in Beijing, China. The concentrations of 16 PAHs and 31 n-alkanes were measured by gas chromatography-mass spectrometry. The total concentrations of PAHs and n-alkanes ranged from 973 ± 55 to 2667 ± 183 μg/kg and 6.40 ± 0.38 to 8.65 ± 0.59 mg/kg (dry weight), respectively, which increased with depth. According to the observed molecular indices, PAHs and n-alkanes originated mostly from petroleum-related sources. The levels of ΣPAHs and the total toxic benzo[a]pyrene equivalent (ranging from 6.41 to 72.54 μg/kg) might exert adverse biological effects. Shotgun metagenomic sequencing was employed to investigate the indigenous microbial community structure and function. The results revealed that Proteobacteria and Actinobacteria were the most abundant phyla, and Nocardioides and Microbacterium were the important genera. Based on COG and KEGG annotations, the highly abundant functional classes were identified, and these functions were involved in allowing microorganisms to adapt to the pressure from contaminants. Five petroleum hydrocarbon degradation-related genes were annotated, revealing the distribution of degrading microorganisms. This work facilitates the understanding of the composition, source, and potential ecological impacts of residual PAHs and n-alkanes in historically contaminated soil.

Bioremediation, which has several advantages over traditional methods, represents an alternative means of dealing with heavy metal pollution. We screened for microorganisms showing heavy metal tolerance in polluted mangrove soils. A novel yeast, Geotrichum sp. CS-67, was discovered and tested for tolerance of Cu

Heavy metal tolerance of microorganisms is the basis of heavy metal removal by growing cells. In this study, a cross-protection effect generated by salt stress significantly enhanced the cadmium tolerance of multi-stress-tolerant Pichia kudriavzevii. Comparative transcriptome analysis using RNA-Seq linked with physiological and biochemical observation was used to elucidate the underlying mechanisms of the improved cadmium tolerance. The expression of cadmium transport related genes (GSTY2, GLR1, GLO2, YCF1 and YOR1), GSH content and GST activity were elevated by salt stress, suggesting enhanced cadmium conjugation and detoxification in yeast cells. The inhibited cadmium uptake by ZRT1 and enhanced cadmium efflux by YOR1 contributed to the decrease in the intracellular cadmium concentration. The improved expression of antioxidant enzyme genes (SOD1, SOD2, SOD6, CAT1 and PRXIID), along with the enhanced activities of antioxidant enzymes (SOD, CAT and POD) resulted in a decrease in cadmium-induced ROS production, protein carbonylation, lipid peroxidation and cell death. The abundant expression of heat shock protein genes (HSP12, HSP10 and SSC1) and genes related to trehalose synthesis (TPS1 and TSL1) induced by salt stress protected yeast cells against complex stress conditions, contributing to the improved cadmium tolerance. These findings will be useful to develop cadmium-tolerant yeasts for cadmium removal by growing cells.

To study the effect of saline-alkali stress on dry mass, rhizosphere soil enzyme activities and soil microbial quantities, pot experiments were designed two sugar beet varieties, 'KWS0143' (strong tolerance to saline-alkali) and 'Beta464' (weak tolerance to saline-alkali) planted in different Na以两个甜菜品种‘KWS0143’(耐盐碱性强)和‘Beta464’(耐盐碱性较弱)为对象,设置4个Na

The remobilization of cadmium (Cd) from shoots to grain is the key process to determine the Cd accumulation in grain. The apoplastic pH of plants is an important factor and signal in influencing on plant responding to environmental variation and inorganic elements uptake. It is proposed that pH of rice plants responds and influences on Cd remobilization from shoots to grain when rice is exposed to Cd stress. The results of hydroponic experiment showed that: pH of the rice leaf vascular bundles among 3 cultivars was almost increased, pH value of 1 cultivar was slightly increasing when rice plants were treated with Cd. The decrease degree of H+ concentration in leaf vascular bundles was different among cultivars. The cultivar with higher decreasing in H+ concentration, showed higher Cd transfer efficiency from shoots to grain. The H+ concentration of leaf vascular bundles under normal condition was negatively correlated to cadmium accumulation in leaf. Moreover, pH change was related to Cd accumulation in shots and remobilization from shoots to grain. Uncovering the role of pH response is a key component for the understanding Cd uptake and remobilization mechanism for rice production.

Soil contaminated by polychlorinated biphenyls (PCBs) is a ubiquitous problem in the world, which can cause significant risks to human health and the environment. Mechanochemical destruction (MCD) has been recognized as a promising technology for the destruction of persistent organic pollutants (POPs) and other organic molecules in both solid waste and contaminated soil. However, few studies have been published about the application of MCD technology for the remediation of PCB contaminated soil. In the present study, the feasibility of destroying PCBs in contaminated soil by co-grinding with and without additives in a planetary ball mill was investigated. After 4 h milling time, more than 96% of PCBs in contaminated soil samples were destroyed. The residual concentrations of PCBs decreased from 1000 mg/kg to below the provisional Basel Convention limit of less than 50 mg/kg. PCDD/F present in the original soil at levels of 4200 ng TEQ/kg was also destroyed with even a slightly higher destruction efficiency. Only minor dechlorinations of the PCBs were observed and the destruction of the hydrocarbon skeleton is proposed as the main degradation pathway of PCBs.

Composting along with rhizodegradation was used to remediate petroleum-contaminated soils in the Yellow River Delta, China. The average concentration of total petroleum hydrocarbons (TPH) in these soils was reduced from 7900-17,900 mg kg(-1) to 1400-3700 mg kg(-1) after field composting. The best volume ratio of amendment to contaminated soil was 2/1 and the best C/N ratio was 15/1. After composting, four local dominant plant species, Seepweed, Sealavander, Central Asia Saltbush and Reed, were selected and planted in composted soils for rhizodegradation in the field. After 90 days of cultivation, the highest net TPH degradation rate was over 40% for Seepweed, probably because of strong root system and active microbial community. In addition, Seepweed roots significantly reduced the surface and volume of soil micropores (which are able to sequestrate organic compounds inside), thus increasing the bioavailability of TPH. In sum, composting followed with planting Seepweed was most effective in remediating the contaminated soil in the Yellow River Delta.

The role of rhizosphere bacteria in facilitating the solubility of copper (Cu) in contaminated soil and Cu accumulation in plant were studied. The bacteria strains were isolated from the rhizosphere of Elsholtzia splendens, a Cu accumulator growing on Tonglu Mountain copper mines. After the sandy soils containing 237 mg kg−1 were incubated with the bacteria strains, it was indicated that rhizosphere microbes played an important role in influencing the availability of water-soluble Cu in soils. Soils had greater concentrations of water-extractable Cu compared with axenic soils inoculated with different bacterial strains. Further evidence for bacterial facilitation of increased solubility of Cu in the soil was obtained using the antibiotic ampicillin (0.1 mg g−1). There were 36% decreases in Cu concentration in the presence of bacterial strain MS12 and ampicillin together compared with bacterial inoculation alone. Different bacterial strains had different abilities on soil water-soluble Cu. To achieve the highest rates of plant Cu accumulation, it was necessary for bacteria to be present in the rhizosphere of E. splendens. Inoculated plants supplied with 20 μmol L−1 CuSO4 had significantly greater concentrations of Cu in shoots and roots than uninoculated plants and bacterial strain MS2 was the most effective strain in promoting plant Cu uptake. There were 2.2-fold and 2.5-fold increases in Cu accumulation in the shoots and roots of plants inoculated with strain MS2 compared to axenic controls. Furthermore, when ampicillin and the bacterial strains were added together to the nutrient solution, the Cu concentrations in roots and shoots of ampicillin-treated plants were lower than those in inoculated plants. When ampicillin was added to the nutrient solution, Cu accumulation was inhibited by about 24–44% in shoots and 20–44% in roots. The above results provided a new insight into the phytoremediation of Cu-contaminated soil. Keywords: Phytoremediation, Soil bacteria, Copper, Mobilization

Soil microbial indicator is one of the important biological indicators in evaluating the extent of soil contamination. In recent years, with the development of molecular biology, many studies have focused on the ecological functions of soil microorganisms by using metagenomics, metatranscriptome and metaproteomics. Relative to metagenomics and metatranscriptome, soil metaproteomics aims to investigate the spatial and temporal changes of the proteins extracted from soil as well as the functional components of soil microbial genomic expression products, which is more conclusive to explore the ecological functions of soil microbes and their roles in soil pollutants transportation and transformation. Therefore, soil metaproteomics has great potential in soil pollution assessment. Currently, soil metaproteomics is still at its infancy stage, while soil protein extraction method is one of the key factors restraining the potential application of soil metaproteomics. In this paper, the advantages and disadvantage of soil metaproteomics in soil pollution assessment were reviewed, with the focus on the comparison of different soil protein extraction methods. In combining with case studies, the feasibility and limits of soil proteins as an indicator for soil pollution assessment were analyzed. In addition, the future research perspectives on the development of soil metaproteomics were discussed.

Arsenic pollution seriously threatens human health and environment safety, being a very prominent environmental issue to be urgently solved in the world. In natural environment and soil systems, arsenic exists in complicated forms, but the plant arsenic poisoning is mainly from As(V) and As (III) exposure. As(V) can be absorbed by plant roots through Pi channel, and reduced rapidly to As(III) by arsenate reductase. As( III) can be transported into plants through NIP channel, and subsequently either transformed into methylated arsenic by arsenic methyltransferase or chelated with the thiol of GSH and PCs. These arsenic compounds can be sequestrated in root cell vacuoles or transported to plant aerial parts. Meanwhile, a part of absorbed arsenic can be discharged to external media. All of these can help to detoxify the arsenic in plants. This paper reviewed the latest research progress on the arsenic- resistance of crops, especially rice, with the focus on the mechanisms of As(V) and As(Ill) absorption and excretion, As(V) reduction, and As (II) methylation and chelation. The major topics of future research on arsenic toxicity were proposed.

The soil of 0-100 cm depth was collected from the wastewater-irrigated farmland in Zhangshi, Shenyang and was amended with low concentration of arsenic. Microbial-mediated speciation transformation and environmental behavior of arsenic were investigated with and without addition of sulfate. The results showed that without addition of sulfate, arsenate was significantly reduced and released into soil solution after eight days of anaerobic incubation. Above 70% of arsenic presented as arsenite. More arsenic was released from the soil of 20-40 cm depth with arsenite and total arsenicconcentration of 892.8 microg x L(-1) and 1 240.6 microg x L(-1) respectively. Compared with abiotic control, the amount of arsenic dissolved in hydrochloric acid decreased greatly in each layer of soil, moreover, almost all of arsenic was reduced to arsenite. Ferric iron was also significantly reduced to ferrous and released into soil solution simultaneously. The concentration of ferrous iron in soil solution was above 40 mg x L(-1) in solution and was 9.0-13.4 g x kg(-1) in soil solid. Above 50% of the iron dissolved in hydrochloric acid was presented as ferrous. This indicates that microbial reduction leads to reductive dissolution of iron oxides and transformation of mineral configuration in soil solid. The release of arsenic and iron was notably suppressed after addition of 10 mmol x L(-1) sulfate, with the concentration reduced by 50%. The amount of HCl-dissolvable arsenic in soil solid decreased by 50%, compared to the treatment without sulfate addition, which probably due to precipitation of arsenic sulfide. It is obvious that microbial reduction leads to reduction and release of arsenic when sulfate is absent. In the presence of sulfate, microbes may transform mobile arsenic into more stable species. Formation of arsenic sulfide mineral was probably the mechanism for arsenic fixation in soil by microbes.

The present study reports the effect of bioaugmentation by free and immobilized bacterial culture on the rhizodegradation of petroleum-polluted soil using Sesbania cannabina plant. Total petroleum hydrocarbon (TPH), hydrocarbon-degrading bacterial counts, microbial activity and root morphology were assessed during 120 days of plant growth. TPH concentration analyzed by GC–MS showed that bioaugmentation did not improve the TPH degradation. TPH concentration decreased from 2541 mg kg−1 to 673 mg kg−1 and 867 mg kg−1 in the rhizosphere of free (FR) and immobilized bacterial inoculated (IR) soil, respectively at the 120th day while in the rhizosphere of uninoculated soil (CR) concentration decreased to 679 mg kg−1 only at the 90th day, showing higher and rapid rhizodegradation with indigenous bacteria than bioaugmented bacterial cultures. Various predominant bacterial groups responsible for higher TPH degradation in the rhizosphere of S. cannabina were identified by PCR–DGGE analysis. It is concluded that natural plant-microbe interaction in the rhizosphere of S. cannabina was efficient enough to degrade TPH and plant rhizosphere keeps bacterial community in its surrounding therefore immobilized culture had no obvious effect on petroleum degradation.

Solid state phosphorous is an important phosphorus speciation controlling the phosphorous bioavailability and mobility in the environment. Synchrotron-based X-ray absorption near-edge structure (XANES) spectroscopy, with its unique advantage of being a non-destructive method in identifying the local chemical information of target element at molecular level in situ, has become one of the frontier technologies for characterizing the speciation of chemical substances and clarifying the microscopic mechanisms of chemical reactions, attracting extensive attention in the field of environmental chemistry. This paper briefly introduced the basic theory of phosphorus XANES spectroscopy, summarized the applications of XANES spectroscopy in the researches of solid state phosphorus speciation in minerals, soils, and organic fertilizers, and analyzed the challenges and prospects of the applications of XANES spectroscopy in identifying the solid state phosphorus speciation in the environmental samples. It was pointed out that XANES spectroscopy should combine with other microscopic spectroscopic techniques and macroscopic analytical methods, complementing each other, to provide a comprehensive and effective technical support for the research of the speciation characterization and transformation mechanisms of phosphorus in the environment.

A hydroponics experiment combined with subcellular fractionation and sequential extraction was conducted to study the Pb concentration in different organs of two tea plant varieties (Longjing 43 and Yingshuang) and the Pb subcellular distribution and chemical forms in the roots of the varieties. Under Pb stress, the root system of the two varieties had different features in morphology. With the increasing concentration of Pb in culture solution, the Pb concentration in Longjing 43 young leaves increased, but that in Yingshuang' s had no significant variation. A marked difference was observed in the Pb subcellular distribution and its chemical forms in roots between the two varieties under Pb stress. In Longjing 43 roots, all subcellular fractions except soluble ones had a lower Pb concentration at low Pb stress, and all the subcellular fractions except cell wall ones had a higher Pb concentration at higher Pb stress, compared with those in Yingshuang's. In Longjing 43 roots, the HAc-extractable Pb occupied the greatest proportion, followed by NaCl-extractable Pb, HCl- and H2O- extractable Pb, and ethanol-extractable Pb; while in Yingshuang's, NaCl-extractable Pb had the greatest proportion, followed by HAc-extractable Pb, HCl- and H2O-extractable Pb, and ethanol-extractable Pb. Based on these findings, tea plant variety Yingshuang was likely to possess a higher tolerance to Pb than Longjing 43 did.

The role of sulfur on the availability of Cu and the bacterial community in rice rhizospheres was investigated by pot experiments. With sulfur addition, pH in rhizosphere soil decreased and Mg(NO3)2 extractable Cu increased significantly. The bacterial community composition also changed with sulfur addition. Some specific clones having high similarity to Thiobacillus, which indicated that sulfur oxidation in the rice rhizosphere could increase the availability of Cu. These results suggested that sulfur source which could provide substrate to sulfur oxidizing bacteria and enhance the availability of Cu was not a suitable sulfur fertilizer for Cu polluted soil.

A pot experiment was conducted to study the microbial activity and community structure in a lead-polluted paddy soil under effects of amendment with different concentration sodium thiosulfate. The amendment of the sulfur-containing substrate increased the soil oxidation-reduction potential and respiration rate, promoted the growth of soil sulfur-oxidizing bacteria, and induced some changes in soil microbial community structure. The clone sequencing indicated that the specific bands of soil microbes in sodium thiosulfate treatments had very high similarity to Bateroidetes, Thiobacillus, beta-proteobacteria, and Acidobacteria. Significant changes were observed in the contents of soil carbonate- and Fe/Mn oxide- bound Pb after the amendment of sodium thiosulfate.

Laboratory simulation tests were conducted to examine the effects of different washing reagents (distilled water, HCl, H3PO4, oxalic acid, and CaCl2) in extracting the heavy metals from contaminated soils left by a chemical plant. The effects of reagent concentration, reaction time, and washing time on the washing efficiency were investigated, and the form variation of test heavy metals was determined before and after HCl washing. Distilled water, H3PO4, and CaCl2 could remove less than 1% of most heavy metals, and the highest removal rate was only 3.58%; while 2 mol HCl x L(-1) could obtain the highest washing efficiency under the optimal conditions, i. e., soil:liquid ratio was 1:3, reaction time was 1 hour, and the soils were washed twice by HCl solution. The removal rates of Cr, Pb, Zn, Cu, and Cd from test soils were 80.75%, 88.69%, 98.00%, 79.33%, and 95.52%, respectively. Among the washing reagents, HCl could effectively remove all forms of heavy metals.

The effects of bio-charcoal acted as sludge-composting additive on soil characteristics and plant growth were studied. Compared with the treatment of composted sludge without bio-charcoal, soil cation exchange capacity in treatment of composted sludge with bio-charcoal increased over 5% and 10% respectively and soil nitrogen content increased 13% and 18% respectively for two kind soils. The composted sludge with bio-charcoal also resulted in 23% higher enhancement on ryegrass biomass and 8%-10% higher enhancement on ryegrass chlorophyll content. In addition, with the amendment of bio-charcoal, the bioavailability of polycyclic aromatic hydrocarbons (PAHs) in composted sludge was decreased, which resulted in the lower absorption and accumulation of ryegrass to PAHs. Compared with the control, the PAHs concentration in ryegrass amended composted sludge with bio-charcoal decreased 27%-34%. The results indicated that composted sludge with bio-charcoal resulted in much more improvement on the plant growth as well as less negative effect on environment. Therefore, biocharcoal was in favor of the safe land application of sewage sludge.

PCB levels in environmental, food, and human samples were determined around a highly PCB-contaminated town (F town), a less contaminated town (P town), and a control town (DXG). There were significant differences in PCB concentrations in the intravenous blood of the mothers and their children living in F, P, and DXG sites. In F town, PCB concentrations in the blood of the mothers averaged 190 mug/kg lipid, as compared to 97 mug/kg lipid in the control site. PCB concentrations in the blood of boys and girls averaged 222 and 153 mug/kg lipid, respectively, in F town. PCB concentrations in the umbilical cord blood averaged 566.8 and 168.2 mug/kg lipid, respectively, in the more seriously (F town) and less seriously (P town) polluted sites. Concentrations in the fetal excreta of the newborns averaged 100.06 and 1.66 mug/kg lipid, respectively, in these sites. PCBs were detected in surface water, underground water, soil, and vegetables, and have spread in a circumference of 30 km from the PCB source and have accumulated in the food chains from surface water to fish and duck eggs and from soil to chickens and pigs. The inhabitants have taken in PCBs through these foods since the 1970s. The results implicate the e-waste recycling operations as having caused elevated PCB levels in the environment and in humans. The elevated exposure levels may have health implications for the next generation.

Using a multi-layer soil column device, the effect of new nitrification inhibitor DMPP (3,4-dimethyl pyrazole phosphate) on nitrogen leaching was studied for understanding the nitrogen vertical transformation and lowering the nitrogen leaching losses. The results indicate that, within 60 days of experiment, the regular urea added with 1% of DMPP can effectively inhibit the ammonium oxidation in the soil, and improve the concentration of NH4(+) -N in soil solution over the 20 cm tilth profile, while decline the concentrations of NO3(-) -N and N2(-) -N. No obvious difference is found on NH4(+) -N concentrations collected from deep layer soil solution under 20 cm between regular urea and the urea added with 1% of DMPP. There is also no significant difference for the NH4(+) -N and NO3(-) -N in the soil solution of deep layer under 40 cm among the treatments of urea by adding with 1% of DMPP within 60 days. So DMPP could be used as an effective nitrification inhibitor to control ammonium oxidation, decline the nitrate leaching losses, minimize the underground water pollution risk and be beneficial for the ecological environment.

Phytolacca acinosa Roxb. (P. acinosa) is a recently discovered manganese hyperaccumulator plant from southern China. It is a good candidate for phytoremediation of manganese(Mn) polluted soil for its high biomass and fast growth. Knowledge of the tissue localization and identification of heavy metals can provide essential information on metal toxicity and bioaccumulation mechanisms. Synchrotron radiation X-ray fluorescence spectroscopy (SRXRF) microprobe was used in this study to investigate the cellular distributions of Mn and other elements in root, stem, leaf, petiole and midrib of P. acinosa. The highest Mn content was found in the vascular tissues of root, stem, petiole and midrib. Cortex in root played a key role in Mn absorption and Mn was limited in the vascular bundle during the process of transportation in stem. Moreover, Mn content in leaf epidermis was higher than that in mesophyll, which suggested that the sequestration of Mn in leaf epidermis might be one of the detoxification mechanisms of P. acinosa. The significance of other elemental (such as P, S, K, Ca, Fe, Zn and Cu) distribution patterns and the correlation with Mn were also discussed.

Phytoremediation has long been recognized as a cost-effective method for the removal of polycyclic aromatic hydrocarbons (PAHs) from soil. A study was conducted to investigate the uptake and accumulation of PAHs in root and shoot of Lolium perenne L. Pot experiments were conducted with series of concentrations of 3.31-378.37 mg/kg for phenanthrene and those of 4.22-365.38 mg/kg for pyrene in a greenhouse. The results showed that both ryegrass roots and shoots did take up PAHs from spiked soils, and generally increased with increasing concentrations of PAH in soil. Bioconcentration factors (BCFs) of phenanthrene by shoots and roots were 0.24--4.25 and 0.17-2.12 for the same treatment. BCFs of pyrene by shoots were 0.20--1.5, except for 4.06 in 4.32 mg/kg treatment, much lower than BCFs of pyrene by roots (0.58--2.28). BCFs of phenanthrene and pyrene tended to decrease with increasing concentrations of phenanthrene and pyrene in soil. Direct uptake and accumulation of these compounds by Lolium perenne L. was very low compared with the other loss pathways, which meant that plant-promoted microbial biodegradation might be the main contribution to plant-enhanced removal of phenanthrene and pyrene in soil. However, the presence of Lolium perenne L. significantly enhanced the removal of phenanthrene and pyrene in spiked soil. At the end of 60 d experiment, the extractable concentrations of phenanthrene and pyrene were lower in planted soil than in non-planted soil, about 83.24%--91.98% of phenanthrene and 68.53%-84.10% of pyrene were removed from soils, respectively. The results indicated that the removal of PAHs in contaminated soils was a feasible approach by using Lolium perenne L.

Humic substrates are a major fraction of sediment organic matters, and the sorption of hydrophobic organic chemicals by humic substrates influences their behavior and fate in sediment. In this paper, organic matters were divided into non-humic substrates and humic substrates. Non-humic substrates include acid leaching fraction, acid extracted fraction, and lipid; humic substrates were fractionated into Ca-binding-FA (fulvic acid), Ca-binding-HA (humic acid), oxide-FA, oxide-HA, and humin. To study the effect of organic fractions on sorption properties, sorption kinetic and equilibrium sorption experiments of phenanthrene and pentachlorophenol (PCP) in five sediments were carried out. The results showed that the contents of acid leaching fraction and oxide-binding-HA were the main fractions to affect the sorption rate constant, and for the sorption capacity of phenanthrene, humin was the major fraction, followed by oxide-binding-HA, oxide-binding-FA, and so on. While for PCP, the factors of influence on sorption capacity were mainly CEC, Ca-binding-FA, and Ca-binding-HA.

A greenhouse experiment was carried out to compare differences in potential activities of ammonification, nitrification and denitrification in rhizosphere and bulk soil in a heavy-metal-stressed system. Exchangeable fractions of Cd, Cu and Cr were all higher in the rhizosphere of maize than in bulk soil. Results showed that the mineralization of N in soil was stimulated by low concentration of Cd. Addition of Cd at low levels stimulated the ammonifying and nitrifying activity in soil, while inhibitory influences were shown at high levels. Nitrifying bacteria was proved to be the most sensitive one, whilst the effect on denitrifying bacteria was very limited. Comparing Cd, Cu and Cr(VI) at 20 mg/kg soil, Cd was the most effective inhibitor of ammonification and denitrification, while Cr(VI) had the strongest inhibitory influence on nitrifying activity. Root exudates played important roles on the different exchangeable metal fractions and bacterial activities between rhizosphere and non-rhizosphere. Nitrate was the main form of mineral N in soil, as well as the main form of N absorbed by plants, but the formation and relative absorption of ammonium were promoted in response to high Cd exposure.