Non-hyperaccumulating Ecotype Research Articles

Cadmium Exposure-Sedum alfredii Planting Interactions Shape the Bacterial Community in the Hyperaccumulator Plant Rhizosphere.

Rhizospheric bacteria play important roles in plant tolerance and activation of heavy metals. Understanding the bacterial rhizobiome of hyperaccumulators may contribute to the development of optimized phytoextraction for metal-polluted soils. We used 16S rRNA gene amplicon sequencing to investigate the rhizospheric bacterial communities of the cadmium (Cd) hyperaccumulating ecotype (HE) Sedum alfredii in comparison to its nonhyperaccumulating ecotype (NHE). Both planting of two ecotypes of S. alfredii and elevated Cd levels significantly decreased bacterial alpha-diversity and altered bacterial community structure in soils. The HE rhizosphere harbored a unique bacterial community differing from those in its bulk soil and NHE counterparts. Several key taxa from Actinobacteria, Bacteroidetes, and TM7 were especially abundant in HE rhizospheres under high Cd stress. The actinobacterial genus Streptomyces was responsible for the majority of the divergence of bacterial community composition between the HE rhizosphere and other soil samples. In the HE rhizosphere, the abundance of Streptomyces was 3.31- to 16.45-fold higher than that in other samples under high Cd stress. These results suggested that both the presence of the hyperaccumulator S. alfredii and Cd exposure select for a specialized rhizosphere bacterial community during phytoextraction of Cd-contaminated soils and that key taxa, such as the species affiliated with the genus Streptomyces, may play an important role in metal hyperaccumulation.IMPORTANCESedum alfredii is a well-known Cd hyperaccumulator native to China. Its potential for extracting Cd relies not only on its powerful uptake, translocation, and tolerance for Cd but also on processes underground (especially rhizosphere microbes) that facilitate root uptake and tolerance of the metal. In this study, a high-throughput sequencing approach was applied to gain insight into the soil-plant-microbe interactions that may influence Cd accumulation in the hyperaccumulator S. alfredii Here, we report the investigation of rhizosphere bacterial communities of S. alfredii in phytoremediation of different levels of Cd contamination in soils. Moreover, some key taxa in its rhizosphere identified in the study, such as the species affiliated with genus Streptomyces, may shed new light on the involvement of bacteria in phytoextraction of contaminated soils and provide new materials for phytoremediation optimization.

A Genetic Transformation Method for Cadmium Hyperaccumulator Sedum plumbizincicola and Non-hyperaccumulating Ecotype of Sedum alfredii

The present study demonstrates the development of an Agrobacterium-mediated genetic transformation method for species of the Sedum genus, which includes the Cd/Zn hyperaccumulator Sedum plumbizincicola and the non-hyperaccumulating ecotype of S. alfredii. Multiple shoots were induced from stem nodes of two Sedum plants using Murashige and Skoog (MS) medium containing 0.1 mg/L cytokinin 6-benzyladenine (6-BA) and 1.0 mg/L auxin 1-naphthaleneacetic acid (NAA). The shoot primordia were used as direct targets for Agrobacterium infection. Selection on hygromycin was highly effective in generating Agrobacterium-transformed explants. This callus-free procedure allowed us to obtain transgenic plantlets after rooting hygromycin-resistant shoots on phytohormone-free MS medium containing the antibiotic. The presence and expression of the reporter genes gusA and GFP in transgenic plants were confirmed by a real-time polymerase chain reaction, histochemical GUS assays, and confocal microscopy. This reliable method for genetic transformation of Sedum plants will help us to understand gene functions and the molecular mechanisms underlying Cd hypertolerance and hyperaccumulation in these species.

Heavy metal ATPase 3 (HMA3) confers cadmium hypertolerance on the cadmium/zinc hyperaccumulator Sedum plumbizincicola.

Cadmium (Cd) is highly toxic to most organisms, but some rare plant species can hyperaccumulate Cd in aboveground tissues without suffering from toxicity. The mechanism underlying Cd detoxification by hyperaccumulators is interesting but unclear. Here, the heavy metal ATPase 3 (SpHMA3) gene responsible for Cd detoxification was isolated from the Cd/zinc (Zn) hyperaccumulator Sedum plumbizincicola. RNA interference (RNAi)-mediated silencing and overexpression of SpHMA3 were induced to investigate its physiological functions in S.plumbizincicola and a nonhyperaccumulating ecotype of Sedum alfredii. Heterologous expression of SpHMA3 in Saccharomyces cerevisiae showed Cd-specific transport activity. SpHMA3 was highly expressed in the shoots and the protein was localized to the tonoplast. The SpHMA3-RNAi lines were hypersensitive to Cd but not to Zn, with the growth of shoots and young leaves being severely inhibited by Cd. Overexpressing SpHMA3 in the nonhyperaccumulating ecotype of S.alfredii greatly increased its tolerance to and accumulation of Cd, but not Zn. These results indicate that elevated expression of the tonoplast-localized SpHMA3 in the shoots plays an essential role in Cd detoxification, which contributes to the maintenance of the normal growth of young leaves of S.plumbizincicola in Cd-contaminated soils.

Unique Rhizosphere Micro-characteristics Facilitate Phytoextraction of Multiple Metals in Soil by the Hyperaccumulating Plant Sedum alfredii.

Understanding the strategies that the roots of hyperaccumulating plants use to extract heavy metals from soils is important for optimizing phytoremediation. The rhizosphere characteristics of Sedum alfredii, a hyperaccumulator, were investigated 6 months after it had been planted in weathered field soils contaminated with 5.8 μg of Cd g-1, 1985.1 μg of Zn g-1, 667.5 μg of Pb g-1, and 698.8 μg of Cu g-1. In contrast with the non-hyperaccumulating ecotype (NHE), the hyperaccumulating ecotype (HE) of S. alfredii was more tolerant to the metals, and higher levels of Cd and Zn accumulated. The HE was characterized by a unique rhizosphere, including extensive root systems, a reduced soil pH, a higher metal bioavailability, and increased rhizomicrobial activity. The bioavailability of metals was significantly correlated with the HE's unique bacterial communities (P < 0.005). The HE harbored abundant Streptomyces (9.43%, family Streptomycetaceae), Kribbella (1.08%, family Nocardioidaceae), and an unclassified genus (1.09%, family Nocardioidaceae) in its rhizosphere, a composition that differed from that of the NHE. PICRUSt analysis predicted high relative abundances of imputed functional profiles in the HE rhizosphere related to membrane transport and amino acid metabolism. This study reveals the rhizosphere characteristics, particularly the unique bacterial rhizobiome of a hyperaccumulator, that might provide a new approach to facilitating heavy metal phytoextraction.

Transcriptome Comparison Reveals the Adaptive Evolution of Two Contrasting Ecotypes of Zn/Cd Hyperaccumulator Sedum alfredii Hance.

Hyperaccumulating ecotype (HE) and non-hyperaccumulating ecotype (NHE) of Sedum alfredii Hance belong to the same species but exhibit contrasting characteristics regarding hyperaccumulation and hypertolerance to cadmium and zinc. The Illumina Hiseq 2500 platform was employed to sequence HE and NHE to study the genetic evolution of this contrasting trait. Greater than 90 million clean reads were obtained and 118,479/228,051 unigenes of HE/NHE were annotated based on seven existing databases. We identified 149,668/319,830 single nucleotide polymorphisms (SNPs) and 12,691/14,428 simple sequence repeats (SSRs) of HE/NHE. We used a branch-site model to identify 18 divergent orthologous genes and 57 conserved orthologous genes of S. alfredii Hance. The divergent orthologous genes were mainly involved in the transcription and translation processes, protein metabolism process, calcium (Ca2+) pathway, stress response process and signal transduction process. To the best of our knowledge, this is the first study to use RNA-seq to compare the genetic evolution of hyperaccumulating and non-hyperaccumulating plants from the same species. In addition, this study made the sole concrete for further studies on molecular markers and divergent orthologous genes to depict the evolution process and formation of the hyperaccumulation and hypertolerance traits in S. alfredii Hance.

The apoplasmic pathway via the root apex and lateral roots contributes to Cd hyperaccumulation in the hyperaccumulator Sedum alfredii.

Although the significance of apoplasmic barriers in roots with regards to the uptake of toxic elements is generally known, the contribution of apoplasmic bypasses (ABs) to cadmium (Cd) hyperaccumulation is little understood. Here, we employed a combination of stable isotopic tracer techniques, an ABs tracer, hydraulic measurements, suberin lamellae staining, metabolic inhibitors, and antitranspirants to investigate and quantify the impact of the ABs on translocation of Cd to the xylem in roots of a hyperaccumulating (H) ecotype and a non-hyperaccumulating (NH) ecotype of Sedum alfredii. In the H ecotype, the Cd content in the xylem sap was proportional to hydrostatic pressure, which was attributed to pressure-driven flow via the ABs. The contribution of the ABs to Cd transportation to the xylem was dependent on the Cd concentration applied to the H ecotype (up to 37% at the highest concentration used). Cd-treated H ecotype roots showed significantly higher hydraulic conductance compared with the NH ecotype (76 vs 52 × 10–8 m s–1MPa–1), which is in accordance with less extensive suberization due to reduced expression of suberin-related genes. The main entry sites of apoplasmically transported Cd were localized in the root apexes and lateral roots of the H ecotype, where suberin lamellae were not well developed. These findings highlight the significance of the apoplasmic bypass in Cd hyperaccumulation in hyperaccumulating ecotypes of S. alfredii.

Enhanced expression of SaHMA3 plays critical roles in Cd hyperaccumulation and hypertolerance in Cd hyperaccumulator Sedum alfredii Hance.

The enhanced expression of a P 1B -type ATPase gene ( SaHMA3 ) is essential for Cd hyperaccumulation and hypertolerance in Sedum alfredii Hance. A functional understanding of the mechanism through which hyperaccumulator plants accumulate and tolerate extremely toxic metals is a prerequisite for the development of novel strategies for improving phytoremediation using engineered plants or natural hyperaccumulators as well as biofortification and food crop safety. Most hyperaccumulator species, however, are small and slow-growing, and their potential for large-scale decontamination of polluted soils is limited. Sedum alfredii Hance, the only one metal hyperaccumulator from the Crassulaceae family, is an ideal candidate for gaining a functional understanding of the intra-family hyperaccumulation mechanisms as well as their potential applications. In the present study, we isolated and functionally characterized a P1B-type ATPase gene (SaHMA3) from S. alfredii Hance. SaHMA3 alleles from a hyperaccumulating ecotype (HE) and non-hyperaccumulating ecotype (NHE) were constitutively expressed in both shoot and root and encoded tonoplast-localized proteins, but showed differences in transport substrate specificity and expression level. SaHMA3 h from the HE plant was a Cd transporter. In contrast, SaHMA3n from NHE plants was able to transport both Zn and Cd. SaHMA3 showed a significantly higher constitutive expression level in HE plants than in NHE plants. Furthermore, the expression level of SaHMA3 in the shoots of HE plants was considerably higher than in the roots. Overexpression of SaHMA3h in tobacco plants significantly enhanced Cd tolerance and accumulation and greatly increased the root sequestration of Cd. In summary, our data suggested that SaHMA3 plays critical roles in Cd hyperaccumulation and hypertolerance in Cd hyperaccumulator S. alfredii Hance.

Functional analysis of CAX2-like transporters isolated from two ecotypes of Sedum alfredii

Two genes encoding CAX2-like proteins were isolated from a Zn/Cd hyperaccumulating ecotype (HE) and nonhyperaccumulating ecotype (NHE) of Sedum alfredii Hance, and they were named SaCAX2h and SaCAX2n, respectively. Both SaCAX2h:eGFP and SaCAX2n:eGFP proteins were localized to the vacuolar membrane of tobacco epidermal cells and yeast mutants. Heterologous expression of SaCAX2h or SaCAX2n in the Δzrc1 yeast mutant increased Cd content in yeast cells. Yeast complementary assay also revealed that both the transporters could suppress Ca and Mn hypersensitivity and enhance Ca and Mn accumulation in a K667 yeast mutant. The expression patterns of the two genes were different under the Cd treatment. Transcription of SaCAX2h was down-regulated in roots and up-regulated in shoots whereas transcription of SaCAX2n was down-regulated in shoots after the exposition to Cd. Furthermore, over-expression of SaCAX2h enhanced metal accumulation in the tobacco plants. The Cd content increased by 17–19 % in shoots and 31–36 % in roots; the Ca content of the transgenic plants increased by 31–32 % in shoots, and the Mn content increased by 60–79 % in shoots and 22–29 % in roots. These results indicate that SaCAX2h was responsible for Ca and Mn sequestration into vacuoles, and over-expression of SaCAX2h enhanced Cd accumulation in the transgenic tobacco.

Oxalate secretion from the root apex of Sedum alfredii contributes to hyperaccumulation of Cd

Organic acids play an important role in metal detoxification in plant, but accumulation of heavy metals with the help of organic acids secretion in hyperaccumulators has not been well documented. The aim of this study was to investigate the contribution of oxalate secretion to cadmium (Cd) hyperaccumulation in S. alfredii. Hydroponic experiments were conducted to investigate the characteristics of Cd-induced secretion of oxalate in terms of pattern, location and oxalate-induced Cd translocation pathway, using ion channel inhibitors, scanning ion-selective electrode technique (SIET) and fluorescence imaging. Hyperaccumulating ecotype (HE) had nearly 2-fold higher oxalate secretion than non-hyperaccumulating ecotype (NHE). Phenylglyoxal effectively blocked Cd-induced oxalate secretion and decreased Cd concentration in HE while exogenous oxalate supply promoted Cd accumulation efficiently. SIET analysis indicated that Cd2+ influx into roots of HE mainly occurred at zone 0–10 mm from root apex where oxalate secretion was localized. Cd was distributed preferentially to the root stele of the HE but not the NHE, and was significantly increased with the secreted oxalates in HE. Cd hyperaccumulation by HE S. alfredii is partially associated with oxalate secretion from the root apex, which mediates the loading of Cd from the xylem parenchyma cells.

Root cell wall polysaccharides are involved in cadmium hyperaccumulation in Sedum alfredii

The role of polysaccharide modification in metal accumulation in hyperaccumulators is still unknown. Our aim was to compare the differences in the role of root cell-wall polysaccharides in cadmium (Cd) accumulation between hyperaccumulating (HE) and non-hyperaccumulating ecotype (NHE) of Sedm alfredii. Hydroponic experiments were performed to characterize root-to-shoot Cd translocation, cadmium species and polysaccharide modification in root cell-wall of S. alfredii using stable isotope tracing, X-ray absorption near edge structure and immunofluorescence localization techniques. Cd absorbed was more readily available for transport to the shoots by the HE roots than by the NHE roots, which is confirmed by a 6-fold higher 113Cd concentration in xylem sap. Root Cd efflux originated mainly from the cell walls. The concentration of cell-wall polysaccharides and activity of pectin methylesterase were higher in the NHE than in the HE in the absence of Cd, and even higher in the presence of Cd. More pectins were methylated in the HE than in the NHE, indicating more free pectic acid residues in the NHE. The cell-wall-bound Cd was retained more tightly in the NHE than in the HE. Cadmium hyperaccumulation by HE of S. alfredii is associated with its enhanced Cd flux into the xylem, which is partly regulated by cell-wall polysaccharide modification in roots.

PsbS expression analysis in two ecotypes of Sedum alfredii and the role of SaPsbS in Cd tolerance

The effects of Cd on the chlorophyll fluorescence parameters of hyperaccumulating ecotype (HE) and non-hyperaccumulating ecotype (NHE) of Sedum alfredii Hance were studied. The photosystem II (PSII) photochemical efficiency of HE plants was not affected, and the non-photochemical quenching (NPQ) value was significantly increased under Cd treatment. In the NHE plants, Cd treatment caused significant loss in PSII photochemical efficiency and NPQ value. Gene expression analysis showed that both ecotypes of S. alfredii accumulated more than twofold higher PsbS (PSII subunit S) transcript levels than control plants after exposure to Cd. Overexpression of PsbS gene isolated from HE plants (SaPsbS) enhanced tobacco growth. The transgenic tobacco possessed greater NPQ capacity in the presence of 100 μM Cd and accumulated the significantly higher Cd content in shoots than the wild-type plants. These data indicate that the SaPsbS may function in NPQ capacity and protect the PSII reaction center in HE plants.

Elevated CO2 concentration increase the mobility of Cd and Zn in the rhizosphere of hyperaccumulator Sedum alfredii

The effects of elevated CO2 on metal species and mobility in the rhizosphere of hyperaccumulator are not well understood. We report an experiment designed to compare the effects of elevated CO2 on Cd/Zn speciation and mobility in the rhizosphere of hyperaccumulating ecotype (HE) and a non-hyperaccumulating ecotype (NHE) of Sedum alfredii grown under ambient (350μll(-1)) or elevated (800μll(-1)) CO2 conditions. No difference in solution pH of NHE was observed between ambient and elevated CO2 treatments. For HE, however, elevated CO2 reduced soil solution pH by 0.22 unit, as compared to ambient CO2 conditions. Elevated CO2 increased dissolved organic carbon (DOC) and organic acid levels in soil solution of both ecotypes, but the increase in HE solution was much greater than in NHE solution. After the growth of HE, the concentrations of Cd and Zn in soil solution decreased significantly regardless of CO2 level. The visual MINTEQ speciation model predicted that Cd/Zn-DOM complexes were the dominant species in soil solutions, followed by free Cd(2+) and Zn(2+) species for both ecotypes. However, Cd/Zn-DOM complexes fraction in soil solution of HE was increased by the elevated CO2 treatment (by 8.01% for Cd and 8.47% for Zn, respectively). Resin equilibration experiment results indicated that DOM derived from the rhizosphere of HE under elevated CO2 (HE-DOM-E) (90% for Cd and 73% for Zn, respectively) showed greater ability to form complexes with Cd and Zn than those under ambient CO2 (HE-DOM-A) (82% for Cd and 61% for Zn, respectively) in the undiluted sample. HE-DOM-E showed greater ability to extract Cd and Zn from soil than HE-DOM-A. It was concluded that elevated CO2 could increase the mobility of Cd and Zn due to the enhanced formation of DOM-metal complexes in the rhizosphere of HE S. alfredii.

Role of sulfur assimilation pathway in cadmium hyperaccumulation by Sedum alfredii Hance

Sedum alfredii Hance is a promising cadmium (Cd) hyperaccumulating plant recently identified in China. However, the physiological and molecular mechanisms underlying Cd accumulation, which differentiate hyperaccumulating ecotype (HE) from non-hyperaccumulating ecotype (NHE) has not been elucidated yet. A hydroponic experiment was conducted to investigate the role of sulfur assimilation pathway in Cd hyperaccumulation by the S. alfredii Hance, by analyzing gene expression pattern in sulfur assimilation pathway and the concentration of some sulfur containing compounds. The results show that, sulfur assimilation pathway was affected by Cd differently in HE and NHE S. alfredii Hance. The gene expression pattern of sulfur assimilation pathway was regulated differently in HE and NHE plants, especially the nicotianamine synthase (NAS). NAS transcript levels in root of HE was 141-fold higher than NHE, while in shoots of HE only 0.31-fold higher than NHE. In HE roots, NAS expression level was maximum 3171-fold higher than shoots, while in NHE plants roots NAS expression level was maximum 45.3-fold higher than shoots. In HE plant roots, sulfur, cysteine and methionine concentrations increased 30%, 46% and 835% respectively, by Cd treatment, but in NHE plants roots, sulfur concentration increased less than 1%, cysteine and methionine concentrations decreased 78.5% and 13.3% respectively, by Cd. Cd exposure increased glutathione levels by 142% in HE but less than 10% in NHE plant roots.

Complexation with dissolved organic matter and mobility control of heavy metals in the rhizosphere of hyperaccumulator Sedum alfredii

The complexation of Zn, Cd and Pb with dissolved organic matter (DOM) in the rhizosphere of hyperaccumulating ecotype (HE) and a non-hyperaccumulating ecotype (NHE) of Sedum alfredii was measured using resin equilibration method. After the growth of HE S. alfredii, the rhizosphere soil pH was reduced by 0.27–0.33 units, due to enhanced DOM derived from root exudation. For both ecotypes of S. alfredii, the fraction of free metal as a percentage of soluble metal varied from 22.1 to 42.5% for Zn2+, from 8.1 to 15.5% for Cd2+, and from 4.5 to 10.4% for Pb2+. Resin equilibration experiment results indicated that HE–DOM had greater ability to form complexes with Zn, Cd and Pb than NHE–DOM, Visual MINTEQ model gave excellent predictions of the complexation of Zn and Cd by DOM (R2 > 0.97). DOM in the rhizosphere of HE S. alfredii could significantly increase metal mobility through the formation of soluble DOM-metal complexes.

Transcriptomic Analysis of Cadmium Stress Response in the Heavy Metal Hyperaccumulator Sedum alfredii Hance

The Sedum alfredii Hance hyperaccumulating ecotype (HE) has the ability to hyperaccumulate cadmium (Cd), as well as zinc (Zn) and lead (Pb) in above-ground tissues. Although many physiological studies have been conducted with these plants, the molecular mechanisms underlying their hyper-tolerance to heavy metals are largely unknown. Here we report on the generation of 9.4 gigabases of adaptor-trimmed raw sequences and the assembly of 57,162 transcript contigs in S. alfredii Hance (HE) shoots by the combination of Roche 454 and Illumina/Solexa deep sequencing technologies. We also have functionally annotated the transcriptome and analyzed the transcriptome changes upon Cd hyperaccumulation in S. alfredii Hance (HE) shoots. There are 110 contigs and 123 contigs that were up-regulated (Fold Change ≧2.0) and down-regulated (Fold Change ≦0.5) by chronic Cd treatment in S. alfredii Hance (HE) at q-value cutoff of 0.005, respectively. Quantitative RT-PCR was employed to compare gene expression patterns between S. alfredii Hance (HE) and non-hyperaccumulating ecotype (NHE). Our results demonstrated that several genes involved in cell wall modification, metal translocation and remobilization were more induced or constitutively expressed at higher levels in HE shoots than that in NHE shoots in response to Cd exposure. Together, our study provides large-scale expressed sequence information and genome-wide transcriptome profiling of Cd responses in S. alfredii Hance (HE) shoots.

Effects of elevated CO2 on rhizosphere characteristics of Cd/Zn hyperaccumulator Sedum alfredii

The effects of elevated CO2 on the metal bioavailability and the rhizosphere characteristics of hyperaccumulator are not well understood. In this study, soil pot experiment was carried out to contrast the effects of elevated CO2 on rhizosphere characteristics between a hyperaccumulating ecotype (HE) and a non-hyperaccumulating ecotype (NHE) of Sedum alfredii grown under ambient (350μLL−1) or elevated (800μLL−1) CO2. Elevated CO2 facilitated the growth of both ecotypes of S. alfredii, but the promotion in the HE was much greater than in the NHE. No significant (P<0.05) changes in soil pH, dissolved organic matter (DOM) and microbial biomass (Cmic) were observed in the rhizosphere of NHE under both CO2 level. For HE, however, elevated CO2 reduced soil pH by 0.3units, increased DOM (especially for hydrophilic acid (HiA) fractions) by 19.2% and Cmic by 19%, as compared to ambient CO2. Mobile Cd and Zn (extractable with 1M NH4NO3) in the rhizosphere of HE decreased considerably, but the decreases were greater under ambient CO2 than under elevated CO2. Phytoextraction efficiency of Cd and Zn by HE was increased significantly by elevated CO2 (P<0.05). The results suggest that elevated CO2 can change soil microenvironment, increase bioavailability of Cd and Zn and thus facilitate metal uptake by the HE. This work highlights that elevated CO2 may be a useful way to improve phytoremediation efficiency of Cd/Zn-contaminated soil by hyperaccumulating ecotype S. alfredii.

Mobilization of cadmium by dissolved organic matter in the rhizosphere of hyperaccumulator Sedum alfredii

Pot experiments were conducted to investigate the role of dissolved organic matter (DOM) in the Cd speciation in the rhizosphere of hyperaccumulating ecotype (HE) and non-hyperaccumulating ecotype (NHE) of Sedum alfredii and its effects on Cd mobility. After growing HE S. alfredii, the rhizosphere soil solution pH of heavily polluted soil (HPS) and slightly polluted soil (SPS) was reduced by 0.49 and 0.40 units, respectively, due to enhanced DOC derived from root exudation. The total Cd concentration in soil solution decreased significantly but the decrease accounted for less than 1% of the total Cd uptake in the shoots of HE S. alfredii. Visual MINTEQ speciation predicted that Cd–DOM complexes were the dominant Cd species in soil solutions after the growth of S. alfredii for both soils, followed by the free metal Cd2+ species. However, Cd–DOM complexes fraction in the rhizosphere soil solution of HE S. alfredii (89.1% and 74.6% for HPS and SPS, respectively) were much greater than NHE S. alfredii (82.8% and 64.7% for HPS and SPS, respectively). Resin equilibration experiment results indicated that DOM from the rhizosphere (R-DOM) of both ecotypes of S. alfredii had the ability to form complexes with Cd, whereas the degree of complexation was significantly higher for HE-R-DOM (79–89%) than NHE-R-DOM (63–74%) in the undiluted sample. The addition of HE-R-DOM significantly (P<0.05) increased the solubility of four Cd minerals while NHE-R-DOM was not as effective at the same concentration. It was concluded that DOM in the rhizosphere of hyperaccumulating ecotype of S. alfredii could significantly increase Cd mobility through the formation of soluble DOM–metal complexes.

Characterization of dissolved organic matter in the rhizosphere of hyperaccumulator Sedum alfredii and its effect on the mobility of zinc

Pot experiments were performed to investigate the characteristics of dissolved organic matter (DOM) in the rhizosphere soil of hyperaccumulating ecotype (HE) and a non-hyperaccumulating ecotype (NHE) of Sedum alfredii and its effects on the mobility of zinc (Zn). DOM was fractionated using XAD resins into six fractions. The acid fraction was the predominant component of DOM in the rhizosphere of S. alfredii, with hydrophilic acid (HiA), hydrophilic base (HiB), and hydrophilic neutral (HiN) in HE-DOM being 1.6, 1.9, and 1.2 times higher respectively, as compared to NHE-DOM. ATR-FTIR results showed that DOM in the rhizosphere of S. alfredii consisted of a mixture of hydroxylated and carboxylic acids, and HE-DOM exhibited more CO, OH, CC and CO functional groups than NHE-DOM. Resin equilibration experiment results indicated that DOM from the rhizosphere of both ecotypes of S. alfredii had the ability to form complexes with Zn, whereas the degree of complexation was significantly higher for HE-DOM (60%) than NHE-DOM (42%). The addition of HE-DOM significantly (P<0.05) increased the solubility of four Zn minerals while NHE-DOM was not as effective at the same concentration. It was concluded that DOM derived from the rhizosphere of hyperaccumulating ecotype of S. alfredii could significantly increase Zn mobility through the formation of soluble DOM–metal complexes, this might be one of the important mechanism by which S. alfredii is involved in activating metal in rhizosphere.

Root adaptations to cadmium-induced oxidative stress contribute to Cd tolerance in the hyperaccumulator Sedum alfredii

Short-term responses of Sedum alfredii roots to Cd exposure was compared in Cd hyperaccumulator (HE) and nonhyperaccumulating ecotype (NHE). Cadmium exposure significantly inhibited root elongation and induced loss of plasma membrane integrity and lipid peroxidation of roots tips in the NHE, whereas these effects were much less pronounced in the HE plants. A strong accumulation of reactive oxygen species with increasing Cd concentration was noted in the NHE root tips, but not in HE. After Cd exposure, a dose-dependent decrease in oxidized glutathione and marked increase in reduced glutathione and non-protein thiols were observed in root tips of HE, but were not seen in the NHE plants. These results suggest that the HE tolerates high Cd in the environment through the differential adaptations against Cd-induced oxidative stress.

Root adaptations to cadmium-induced oxidative stress contribute to Cd tolerance in the hyperaccumulator Sedum alfredii

Short-term responses of Sedum alfredii roots to Cd exposure was compared in Cd hyperaccumulator (HE) and nonhyperaccumulating ecotype (NHE). Cadmium exposure significantly inhibited root elongation and induced loss of plasma membrane integrity and lipid peroxidation of roots tips in the NHE, whereas these effects were much less pronounced in the HE plants. A strong accumulation of reactive oxygen species with increasing Cd concentration was noted in the NHE root tips, but not in HE. After Cd exposure, a dose-dependent decrease in oxidized glutathione and marked increase in reduced glutathione and non-protein thiols were observed in root tips of HE, but were not seen in the NHE plants. These results suggest that the HE tolerates high Cd in the environment through the differential adaptations against Cd-induced oxidative stress.