Does phytoextraction with Sedum plumbizincicola increase cadmium leaching from polluted agricultural soil?

Sedum plumbizincicola is a cadmium (Cd) and zinc hyperaccumulator that can activate Cd by rhizosphere acidification. However, there is little understanding of the Cd leaching risk from polluted soil during phytoextraction process. Here, pot and column experiments were conducted to monitor soil Cd leaching characteristics under different rainfall simulation conditions during S. plumbizincicola phytoextraction. Soil Cd leaching increased significantly with increasing simulated rainfall intensity. Compared with normal rainfall (NR), weak rainfall (WR) resulted in a 34.3% decrease in Cd uptake by S. plumbizincicola and also led to a 68.7% decline in Cd leaching. In contrast, Cd leaching under heavy rainfall (HR) was 2.12 times that of NR in the presence of S. plumbizincicola. After two successive growing periods, phytoextraction resulted in a 53.5-66.4% decline in the amount of soil Cd leached compared with controls in which S. plumbizincicola was absent. Even compared with maize cropping as a control, S. plumbizincicola did not instigate a significant increase in Cd leaching. The contribution of Cd leaching loss to the decline in soil total Cd concentration was negligible after phytoextraction in the pot experiment. Overall, the results contribute to our understanding of soil Cd leaching risk by phytoextraction with S. plumbizincicola.

Repeated phytoextraction by hyperaccumulator Sedum plumbizincicola is an important remediation technology to remove Cd from contaminated soils. At the same time, Sedum plumbizincicola can also activate soil Cd by rhizosphere acidification. However, studies on the leaching risk of soil activated Cd during the phytoextraction process are very few. This study looked at the effects of Sedum plumbizincicola growth on soil Cd leaching with the changes in rainfall simulation and plant type. Results showed that repeated phytoextraction with Sedum plumbizincicola did not increase Cd leaching from contaminated soil.

Exploring Transcriptional Regulation of Hyperaccumulation in Sedum plumbizincicola through Integrated Transcriptome Analysis and CRISPR/Cas9 Technology.

The cadmium hyperaccumulator Sedum plumbizincicola has remarkable abilities for cadmium (Cd) transport, accumulation and detoxification, but the transcriptional regulation mechanisms responsible for its Cd hyperaccumulation remain unknown. To address this knowledge gap, we conducted a comparative transcriptome study between S. plumbizincicola and the non-hyperaccumulating ecotype (NHE) of Sedum alfredii with or without Cd treatment. Our results revealed many differentially expressed genes involved in heavy metal transport and detoxification that were abundantly expressed in S. plumbizincicola. Additionally, we identified a large number of differentially expressed transcription factor genes, highlighting the complexity of transcriptional regulatory networks. We further screened four transcription factor genes that were highly expressed in the roots of S. plumbizincicola as candidate genes for creating CRISPR/Cas9 knockout mutations. Among these, the SpARR11 and SpMYB84 mutant lines exhibited decreased Cd accumulation in their aboveground parts, suggesting that these two transcription factors may play a role in the regulation of the Cd hyperaccumulation in S. plumbizincicola. Although further research will be required to determine the precise targeted genes of these transcription factors, combined transcriptome analysis and CRISPR/Cas9 technology provides unprecedented opportunities for identifying transcription factors related to Cd hyperaccumulation and contributes to the understanding of the transcriptional regulation mechanism of hyperaccumulation in S. plumbizincicola.

Integrative analysis of transcriptome and proteome provides insights into adaptation to cadmium stress in Sedum plumbizincicola.

Sedum plumbizincicola, a cadmium (Cd) hyperaccumulating herbaceous plant, can accumulate large amounts of Cd in the above-ground tissues without being poisoned. However, the molecular mechanisms regulating the processes are not fully understood. In this study, Transcriptional and proteomic analyses were integrated to investigate the response of S. plumbizincicola plants to Cd stress and to identify key pathways that are potentially responsible for Cd tolerance and accumulation. A total of 630 DAPs (differentially abundant proteins, using fold change >1.5 and adjusted p-value <0.05) were identified from Tandem Mass Tag (TMT)- based quantitative proteomic profiling, which were enriched in processes including phenylpropanoid biosynthesis, protein processing in endoplasmic reticulum, and biosynthesis of secondary metabolites. Combined with the previous transcriptomic study, 209 genes and their corresponding proteins showed the identical expression pattern. The identified genes/proteins revealed the potential roles of several metabolism pathways, including phenylpropanoid biosynthesis, oxidative phosphorylation, phagosome, and glutathione metabolism, in mediating Cd tolerance and accumulation. Lignin staining and Cd accumulation assay of the transgenic lines over-expressing a selected Cd up-regulated gene SpFAOMT (Flavonoid 3',5'-methyltransferase) showed its functions in adapting to Cd stress, and provided insight into its role in lignin biosynthesis and Cd accumulation in S. plumbizincicola during Cd stress.

Dispose waste liquor of fresh biomass of a hyperaccumulator Sedum plumbizincicola in phytoextraction process.

Sedum plumbizincicola has been widely employed to remove cadmium (Cd) and zinc (Zn) from contaminated soils and harvested biomass is used to recover valuable metals. While chopping and compacting are efficient methods to rapidly reduce the volume and moisture of fresh biomass, the resulting waste liquor containing metals needs treatment. Two types of contaminated soils were cropped with S. plumbizincicola and top-dressed with this liquor to study metals migration in soil profile and their uptake by plants. There were three treatments: planting and adding liquor (PL), planting without liquor (P) and adding liquor without planting (L). The results showed that Cd and Zn from liquor were mainly retained at top soil 0-10 cm under L treatment. Compared with L treatment, soil Cd and Zn under PL treatment decreased significantly in soil profile due to the extraction of S. plumbizincicola. Moreover, the amount of Cd and Zn extracted by plants was greater than that applied in soils. The metal removal rate by S. plumbizincicola in acid clay loam soil was higher than that in neutral sandy soil. To sum up, metal retaining in soil and uptake by S. plumbizincicola can be used to treat waste liquor from its fresh biomass. Novelty StatementRapid and efficient treatment of harvested fresh biomass is still a challenge although phytoextraction using hyperaccumulator Sedum plumbizincicola has been widely employed. Chopping and compacting fresh biomass are efficient methods for rapid dehydration, however, a large amount of waste liquor that contains of Cd and Zn is produced and needs treatment. In the present study, a simple and low-cost method was tested to dispose the liquor, i. e. irrigating it onto the surface of contaminated soils where grown S. plumbizincicola. It was found that Cd and Zn applied in soils from liquor were mainly retained at top 0-10 cm soil depth where S. plumbizincicola root system was widespread, and the amount of Cd and Zn extracted by plants was greater than that applied in soils. Therefore, it is technically feasible to dispose the waste liquor dewatering from fresh biomass of S. plumbizincicola in its phytoextraction process. This study is helpful for the rapid, efficient and low-cost treatment of harvested fresh biomass in the large-scale application of phytoremediation.

Cadmium uptake and transfer by Sedum plumbizincicola using EDTA, tea saponin, and citric acid as activators.

Sedum plumbizincicola (S. plumbizincicola) is known as a sufficient plant for phytoremediation of cadmium (Cd) polluted soils. This study aimed to investigate the effects of ethylene diamine tetraacetic acid (EDTA), tea saponin (TS), and citric acid (CA) on Cd uptake and translocation by S. plumbizincicola. To do so, using a pot experiment, we set four concentration levels of activators (1, 3, 5, and 10 mmol L-1) and a control (CK). Results showed that none of the applied activators had significant impact on soil pH. Except for CA-10, the concentration of available Cd in Cd polluted soils increased by 65.8-72.9% compared with CK. The EDTA-1, CA-1, and TS-5 treatments caused significant increases of 52.3, 67.2, and 38.4%, respectively, in the biomass of aerial parts of S. plumbizincicola (p < 0.05) compared with CK. Except for CA-3, activators increased Cd accumulation in the aerial parts of plants by 47-124% compared with CK. Of all activators, EDTA-3 caused the highest Cd accumulation of 6.64 g pot-1 in the aerial plant tissues followed by CA-10 (6.25 g pot-1) and TS-1 (5.48 g pot-1). Finally, our results suggested that the application of S. plumbizincicola together with different activators sufficiently reduced soil total Cd by 4.64-48.4% compared with CK. These findings suggest that appropriate application of EDTA, TS, and CA can promote phytoremediation of Cd contaminated soils by hyper-accumulators. In particular, the combined use of EDTA and S. plumbizincicola is an affordable and promising strategy for remediation of Cd contaminated soil.Novelty statement: Sedum plumbizincicola (S. plumbizincicola) is a well-known hyper-accumulator plant for remediation of cadmium (Cd) and zinc (Zn) contaminated soils. In addition, low molecular rganic acids and macromolecular chelating agents can improve the solubility and leaching of soil heavy metals. In the present work, we examined the combined effects of three activators (EDTA, tea saponin, and citric acid) with S. plumbizincicola to remediate a Cd contaminated soil in Anhui Province, East China. Our results indicated the effectiveness of these activators to increase soil available Cd, as well as improving the biomass of S. plumbizincicola and its Cd uptake. We believe that this study provides an efficient approach to increase the uptake of Cd by S. plumbizincicola, restoring Cd contaminated soils. Nevertheless, excessive activators may have adverse effects on soil aggregates and soil microorganisms. Therefore, it is necessary to control the amount of chelating agents and subsequently the deterioration of soil quality.

An electro-Fenton process to treat waste liquor of a hyperaccumulator that contains potentially toxic elements and the COD.

Rapid and safe treatment of harvested fresh biomass of hyperaccumulators is essential for phytoremediation of metal-contaminated soils. Here, an electro-Fenton (EF) process was used to remove cadmium (Cd) and chemical oxidation demand (COD) from waste liquor from the dewatering of biomass of the hyperaccumulator Sedum plumbizincicola after flocculation precipitation. The results showed that the order of impact of the factors on the removal rate of COD and Cd was pH > electrical current density > H2O2 dosage. Increasing pH promoted Cd removal but hindered COD removal. As current density and H2O2 dosage increased the removal rates of both Cd and COD initially increased and then decreased. Compared to an electrocoagulation process, the addition of H2O2 in EF process greatly enhanced Cd and zinc (Zn) removal. Speciation analysis showed that most of the Cd and Zn in the initial liquor were organically and inorganically complexed. At optimal conditions, e.g., pH 5, current density 15 mA cm-2 and H2O2 dosage 9 g L-1, the removal efficiencies of Cd, Zn and COD reached 99.4, 99.9 and 55.5% after 80 min of EF treatment. Electro-Fenton process can therefore be used to quickly remove trace metals from the waste liquor of the hyperaccumulator.

Analysis of the effect of cadmium stress on root exudates of Sedum plumbizincicola based on metabolomics.

Root exudates are the most direct manifestation of the response of plants changes in the external environment. Therefore, based on non-targeted gas chromatography-time-of-flight mass spectrometry and metabolomics, the response of Sedum plumbizincicola root exudates to Cd stress was used to reveal the possible mechanism of resistance to or accumulation of Cd. The results showed that Cd significantly changed the composition and contents of S. plumbizincicola root exudates. A total of 155 metabolites were identified in S. plumbizincicola root exudates, among which 33 showed significant differences under Cd stress, including organic acids, amino acids, lipids, and polyols. Cd stress suppressed organic acid metabolism and lipid metabolism in S. plumbizincicola and significantly affected amino acid metabolism. There were 16 metabolic pathways related to Cd stress, among which arginine and proline metabolism, valine, leucine, and isoleucine biosynthesis, glycine, serine, and threonine metabolism, glutathione metabolism, and purine metabolism were the key pathways with the highest correlation, and were closely related to the stress resistance of S. plumbizincicola.

SpHMA1 is a chloroplast cadmium exporter protecting photochemical reactions in the Cd hyperaccumulator Sedum plumbizincicola.

Sedum plumbizincicola is able to hyperaccumulate cadmium (Cd), a nonessential and highly toxic metal, in the above-ground tissues, but the mechanisms for its Cd hypertolerance are not fully understood. Here, we show that the heavy metal ATPase 1 (SpHMA1) of S. plumbizincicola plays an important role in chloroplast Cd detoxification. Compared with the HMA1 ortholog in the Cd nonhyperaccumulating ecotype of Sedum alfredii, the expression of SpHMA1 in the leaves of S. plumbizincicola was >200 times higher. Heterologous expression of SpHMA1 in Saccharomyces cerevisiae increased Cd sensitivity and Cd transport activity in the yeast cells. The SpHMA1 protein was localized to the chloroplast envelope. SpHMA1 RNA interference transgenic plants and CRISPR/Cas9-induced mutant lines showed significantly increased Cd accumulation in the chloroplasts compared with wild-type plants. Chlorophyll fluorescence imaging analysis revealed that the photosystem II of SpHMA1 knockdown and knockout lines suffered from a much higher degree of Cd toxicity than wild type. Taken together, these results suggest that SpHMA1 functions as a chloroplast Cd exporter and protects photosynthesis by preventing Cd accumulation in the chloroplast in S. plumbizincicola and hyperexpression of SpHMA1 is an important component contributing to Cd hypertolerance in S. plumbizincicola.

Comparison of heavy metal phytoremediation in monoculture and intercropping systems of Phyllostachys praecox and Sedum plumbizincicola in polluted soil.

A bamboo species (Phyllostachys praecox) and a Cd/Zn hyperaccumulator (Sedum plumbizincicola) were tested under different planting systems to compare their heavy metal phytoremediation ability. P. praecox (MP), S. plumbizincicola (MS) and P. praecox × S. plumbizincicola (IPS) plantations were established in Cu, Zn, and Cd-contaminated soil. Soil properties and heavy metal contents in plants were determined and compared after four years of plantation establishment. The rankings of available and total metal contents in soil layers were MP > MS > IPS (0-20 cm) and MP > IPS > MS (20-40 cm, except for Cu), respectively. The Cu and Zn contents in mature bamboo tissues were significantly lower, but the Cd contents in bamboo tissues (except for leaves) higher, in the IPS than in the MP. The bioconcentration and the translocation factors in most of bamboo tissues showed an increasing trend from the MP to the IPS. Heavy metal distribution in plants is greatly affected by the planting patterns. The tested intercropping system of two plant species showed higher biomass productivity, implying more heavy metals can be removed from the soil through the harvesting of plants. Therefore, the IPS leads to significant improvement of soil phytoremediation.

The hyperaccumulator Sedum plumbizincicola harbors metal-resistant endophytic bacteria that improve its phytoextraction capacity in multi-metal contaminated soil.

Endophyte-assisted phytoremediation has recently been suggested as a successful approach for ecological restoration of metal contaminated soils, however little information is available on the influence of endophytic bacteria on the phytoextraction capacity of metal hyperaccumulating plants in multi-metal polluted soils. The aims of our study were to isolate and characterize metal-resistant and 1-aminocyclopropane-1-carboxylate (ACC) utilizing endophytic bacteria from tissues of the newly discovered Zn/Cd hyperaccumulator Sedum plumbizincicola and to examine if these endophytic bacterial strains could improve the efficiency of phytoextraction of multi-metal contaminated soils. Among a collection of 42 metal resistant bacterial strains isolated from the tissues of S. plumbizincicola grown on Pb/Zn mine tailings, five plant growth promoting endophytic bacterial strains (PGPE) were selected due to their ability to promote plant growth and to utilize ACC as the sole nitrogen source. The five isolates were identified as Bacillus pumilus E2S2, Bacillus sp. E1S2, Bacillus sp. E4S1, Achromobacter sp. E4L5 and Stenotrophomonas sp. E1L and subsequent testing revealed that they all exhibited traits associated with plant growth promotion, such as production of indole-3-acetic acid and siderophores and solubilization of phosphorus. These five strains showed high resistance to heavy metals (Cd, Zn and Pb) and various antibiotics. Further, inoculation of these ACC utilizing strains significantly increased the concentrations of water extractable Cd and Zn in soil. Moreover, a pot experiment was conducted to elucidate the effects of inoculating metal-resistant ACC utilizing strains on the growth of S. plumbizincicola and its uptake of Cd, Zn and Pb in multi-metal contaminated soils. Out of the five strains, B. pumilus E2S2 significantly increased root (146%) and shoot (17%) length, fresh (37%) and dry biomass (32%) of S. plumbizincicola as well as plant Cd uptake (43%), whereas Bacillus sp. E1S2 significantly enhanced the accumulation of Zn (18%) in plants compared with non-inoculated controls. The inoculated strains also showed high levels of colonization in rhizosphere and plant tissues. Results demonstrate the potential to improve phytoextraction of soils contaminated with multiple heavy metals by inoculating metal hyperaccumulating plants with their own selected functional endophytic bacterial strains.

Impacts of simulated atmospheric cadmium deposition on the physiological response and cadmium accumulation of Sedum plumbizincicola.

Atmospheric cadmium (Cd) deposition contributes to the accumulation of Cd in the soil-plant system. Sedum plumbizincicola is a Cd and Zn hyperaccumulator commonly used for the phytoremediation of Cd-contaminated soil. However, studies on the effects of atmospheric Cd deposition on the accumulation of Cd and physiological response in S. plumbizincicola are still limited. A Cd solution spraying pot experiment was conducted with S. plumbizincicola at three atmospheric Cd deposition concentrations (4, 8, and 12 mg/L). Each Cd concentration levels was divided into two groups, non-mulching (foliar-root uptake) and mulching (foliar uptake). The soil type used in the experiment was reddish clayey soil collected from a farmland. The results showed that compared with the non-mulching control, the fresh weight of S. plumbizincicola in non-mulching with high atmospheric Cd deposition (12 mg/L) increased by 11.35%. Compared with those in the control group, the malondialdehyde (MDA) content in the non-mulching and mulching S. plumbizincicola groups increased by 0.88-11.06 nmol/L and 0.96-1.32 nmol/L, respectively. Compared with those in the non-Cd-treated control group, the shoot Cd content in the mulching group significantly increased by 11.09-180.51 mg/kg. Under high Cd depositions, the Cd in S. plumbizincicola mainly originated from the air and was stored in the shoots (39.7-158.5%). These findings highlight that the physiological response and Cd accumulation of S. plumbizincicola were mainly affected by high Cd deposition and suggest that atmospheric Cd could directly be absorbed by S. plumbizincicola. The effect of atmospheric deposition on S. plumbizincicola cannot be ignored.

SpSIZ1 from hyperaccumulator Sedum plumbizincicola orchestrates SpABI5 to fine-tune cadmium tolerance.

Sedum plumbizincicola is a renowned hyperaccumulator of cadmium (Cd), possesses significant potential for eco-friendly phytoremediation of soil contaminated with Cd. Nevertheless, comprehension of the mechanisms underpinning its Cd stress response remains constrained, primarily due to the absence of a comprehensive genome sequence and an established genetic transformation system. In this study, we successfully identified a novel protein that specifically responds to Cd stress through early comparative iTRAQ proteome and transcriptome analyses under Cd stress conditions. To further investigate its structure, we employed AlphaFold, a powerful tool for protein structure prediction, and found that this newly identified protein shares a similar structure with Arabidopsis AtSIZ1. Therefore, we named it Sedum plumbizincicola SIZ1 (SpSIZ1). Our study revealed that SpSIZ1 plays a crucial role in positively regulating Cd tolerance through its coordination with SpABI5. Overexpression of SpSIZ1 significantly enhanced plant resistance to Cd stress and reduced Cd accumulation. Expression pattern analysis revealed higher levels of SpSIZ1 expression in roots compared to stems and leaves, with up-regulation under Cd stress induction. Importantly, overexpressing SpSIZ1 resulted in lower Cd translocation factors (Tfs) but maintained relatively constant Cd levels in roots under Cd stress, leading to enhanced Cd stress resistance in plants. Protein interaction analysis revealed that SpSIZ1 interacts with SpABI5, and the expression of genes responsive to abscisic acid (ABA) through SpABI5-dependent signaling was significantly up-regulated in SpSIZ1-overexpressing plants with Cd stress treatment. Collectively, our results illustrate that SpSIZ1 interacts with SpABI5, enhancing the expression of ABA downstream stress-related genes through SpABI5, thereby increasing Cd tolerance in plants.

Rotation of Celosia argentea and Sedum plumbizincicola promotes Cd phytoextraction efficiency.

Most hyperaccumulators cannot maintain vigorous growth throughout the year, which may result in a low phytoextraction efficiency for a few months. In the present study, rotation of two hyperaccumulators is proposed to address this issue. An 18-month field experiment was conducted to evaluate the phytoextraction efficiency of Cd by the monoculture and rotation of Celosia argentea and Sedum plumbizincicola. The results showed that rotation increased amount of extracted Cd increased by 2.3 and 1.6 times compared with monoculture of C. argentea and S. plumbizincicola. In rotation system, the biomass of S. plumbizincicola and Cd accumulation in C. argentea increased by 54.4% and 40.7%, respectively. Rotation reduced fallow time and increased harvesting frequency, thereby enhancing Cd phytoextraction. Planting C. argentea significantly decreased soil pathogenic microbes and increased the abundances of plant growth-promoting rhizobacteria (PGPR) and 1-aminocyclopropane-1-carboxylate (ACC) deaminase genes, which may be beneficial for the growth of S. plumbizincicola. Planting S. plumbizincicola increased the abundance of sulfur oxidization (SOX) system genes and decreased soil pH (p < 0.05), thereby increasing the Cd uptake by C. argentea. These findings indicated that rotation of C. argentea and S. plumbizincicola is a promising method for promoting Cd phytoextraction.

Cadmium levels and soil pH drive structure and function differentiation of endophytic bacterial communities in Sedum plumbizincicola: A field study.

Sedum plumbizincicola is a promising hyperaccumulator for heavy metal phytoremediation. It grows in heavy metal polluted soil and stores specific endophyte resources with heavy metal tolerance or growth promotion characteristics. In this study, the endophyte communities of S. plumbizincicola, growing naturally in the field (two former mining locations and one natural location) were investigated, and their structure and function were comparatively studied. The bioaccumulation and translocation characteristics of cadmium (Cd) and selenium (Se) in S. plumbizincicola were also evaluated. The results showed that the heavy metal pollution reduced the richness and diversity of endophyte communities. Soil pH and Cd concentration could be the key factors affecting the composition of the endophyte community. Co-occurrence network analysis identified that 22 keystone taxa belonging to Actinobacteriota, Firmicutes, Myxococcota and Proteobacteria were positively correlated with Cd bioaccumulation and translocation. The predicted endophyte metabolic pathways were enriched in physiological metabolism, immune system, and genetic Information processing. These findings may help to understand how endophytes assist host plants to enhance their adaptability to harsh environments, and provide a basis for further exploration of plant-endophyte interactions and improvement in phytoremediation efficiency.

SpbZIP60 confers cadmium tolerance by strengthening the root cell wall compartmentalization in Sedum plumbizincicola.

Cadmium (Cd) is a prominent heavy metal pollutant that inhibits plant growth and poses risks to human health. Sedum plumbizincicola, as a Cd/Zn/Pb hyperaccumulator species, exhibits robust resistance to heavy metals and effective enrichment capacities. In our previous study, overexpressing SpbZIP60 in Arabidopsis enhanced Cd tolerance; however, the underlying the molecular mechanism remains to be elucidated. Here, we identified SpbZIP60 as a representative Cd stress response factor with nuclear localization and transcriptional activation activity. SpbZIP60 underwent conservative splicing in response to endoplasmic reticulum (ER) stress, while its response to Cd stress is independent of the ER stress-mediated unfolded protein response pathway. Overexpression of SpbZIP60 in S. alfredii increased the Cd tolerance and antioxidant activity. Furthermore, SpbZIP60 increased the content of cell wall components and thickened cell wall under Cd stress. Transcriptome analysis indicated a significant enrichment of differentially expressed genes within the phenylpropanoid metabolism pathway. Besides, the binding of SpbZIP60 to the promoter region of SpBglu resulted in the activation of gene expression, thereby enhancing the process of lignin deposition. Collectively, our results elucidated a molecular regulatory model in which SpbZIP60 increased the thickness of the root cell wall to impede Cd entry into the cell, consequently improving Cd tolerance.

A novel gene SpCTP3 from the hyperaccumulator Sedum plumbizincicola redistributes cadmium and increases its accumulation in transgenic Populus × canescens.

A cadmium (Cd) tolerance protein (SpCTP3) involved in the Sedum plumbizincicola response to Cd stress was identified. However, the mechanism underlying the Cd detoxification and accumulation mediated by SpCTP3 in plants remains unclear. We compared wild-type (WT) and SpCTP3-overexpressing transgenic poplars in terms of Cd accumulation, physiological indices, and the expression profiles of transporter genes following with 100 µmol/L CdCl2. Compared with the WT, significantly more Cd accumulated in the above-ground and below-ground parts of the SpCTP3-overexpressing lines after 100 µmol/L CdCl2 treatment. The Cd flow rate was significantly higher in the transgenic roots than in the WT roots. The overexpression of SpCTP3 resulted in the subcellular redistribution of Cd, with decreased and increased Cd proportions in the cell wall and the soluble fraction, respectively, in the roots and leaves. Additionally, the accumulation of Cd increased the reactive oxygen species (ROS) content. The activities of three antioxidant enzymes (peroxidase, catalase, and superoxide dismutase) increased significantly in response to Cd stress. The observed increase in the titratable acid content in the cytoplasm might lead to the enhanced chelation of Cd. The genes encoding several transporters related to Cd2+ transport and detoxification were expressed at higher levels in the transgenic poplars than in the WT plants. Our results suggest that overexpressing SpCTP3 in transgenic poplar plants promotes Cd accumulation, modulates Cd distribution and ROS homeostasis, and decreases Cd toxicity via organic acids. In conclusion, genetically modifying plants to overexpress SpCTP3 may be a viable strategy for improving the phytoremediation of Cd-polluted soil.

Overexpression of Sedum SpHMA2, SpHMA3 and SpNramp6 in Brassica napus increases multiple heavy metals accumulation for phytoextraction.

Phytoextraction is an environmentally friendly phytoremediation technology that can reduce the total amount of heavy metals (HMs) in the soil. Hyperaccumulators or hyperaccumulating transgenic plants with biomass are important biomaterials for phytoextraction. In this study, we show that three different HM transporters from the hyperaccumulator Sedum pumbizincicola, SpHMA2, SpHMA3, and SpNramp6, possess Cd transport. These three transporters are located at the plasma membrane, tonoplast, and plasma membrane, respectively. Their transcripts could be strongly stimulated by multiple HMs treatments. To create potential biomaterials for phytoextraction, we overexpressed the three single genes and two combining genes, SpHMA2&SpHMA3 and SpHMA2&SpNramp6, in rapes having high biomass and environmental adaptability, and found that the aerial parts of the SpHMA2-OE3 and SpHMA2&SpNramp6-OE4 lines accumulated more Cd from single Cd-contaminated soil because SpNramp6 transports Cd from root cells to the xylem and SpHMA2 from the stems to the leaves. However, the accumulation of each HM in the aerial parts of all selected transgenic rapes was strengthened in multiple HMs-contaminated soils, probably due to the synergistic transport. The HMs residuals in the soil after the transgenic plant phytoremediation were also greatly reduced. These results provide effective solutions for phytoextraction in both Cd and multiple HMs-contaminated soils.

The Mitogenome of Sedum plumbizincicola (Crassulaceae): Insights into RNA Editing, Lateral Gene Transfer, and Phylogenetic Implications.

As the largest family within the order Saxifragales, Crassulaceae contains about 34 genera with 1400 species. Mitochondria play a critical role in cellular energy production. Since the first land plant mitogenome was reported in Arabidopsis, more than 400 mitogenomic sequences have been deposited in a public database. However, no entire mitogenome data have been available for species of Crassulaceae to date. To better understand the evolutionary history of the organelles of Crassulaceae, we sequenced and performed comprehensive analyses on the mitogenome of Sedum plumbizincicola. The master mitogenomic circle is 212,159 bp in length, including 31 protein-coding genes (PCGs), 14 tRNA genes, and 3 rRNA genes. We further identified totally 508 RNA editing sites in PCGs, and demonstrated that the second codon positions of mitochondrial genes are most prone to RNA editing events. Notably, by neutrality plot analyses, we observed that the mitochondrial RNA editing events have large effects on the driving forces of plant evolution. Additionally, 4 MTPTs and 686 NUMTs were detected in the mitochondrial and nuclear genomes of S. plumbizincicola, respectively. Additionally, we conducted further analyses on gene transfer, secondary structures of mitochondrial RNAs, and phylogenetic implications. Therefore, the findings presented here will be helpful for future investigations on plant mitogenomes.

Effects of a soil collembolan on the growth and metal uptake of a hyperaccumulator: Modification of root morphology and the expression of plant defense genes.

Soil collembolans live in close proximity to plant roots and may have a role in the phytoextraction of potentially toxic metals from contaminated soils but the underlying mechanisms remain poorly investigated. We hypothesize that soil collembolans may change the root morphology of hyperaccumulators by regulating plant physiological characteristics. Here, a pot experiment was conducted in which a cadmium (Cd) and zinc (Zn) hyperaccumulator (Sedum plumbizincicola) was grown with or without a collembolan (Folsomia candida), and plant transcriptome and hormones as well as the root characteristics of S. plumbizincicola were analyzed. F. candida promoted the growth and Cd/Zn uptake of S. plumbizincicola, the root and shoot biomass increasing by 53.3 and 34.4%, and the uptake of Cd and Zn in roots increased by 83.2 and 65.4%, respectively. Plant root morphology, total root length, root tip number and lateral root number increased significantly by 40.7, 37.2 and 33.8%, respectively, with the addition of F. candida. Transcriptome analysis reveals that the expression levels of defense-related genes in S. plumbizincicola were significantly up-regulated. In addition, the defensive plant hormones, i.e. salicylic acid in the roots, increased significantly by 338%. These results suggest that the plant in defense of the action of F. candida regulated the expression of the corresponding genes and increased the defensive plant hormones, thus modifying root morphology and plant performance. Overall, this study highlights the importance of the regulation by collembolans of plant growth and metal uptake by interaction with hyperaccumulator roots.

Effects of four endophytic bacteria on cadmium speciation and remediation efficiency of Sedum plumbizincicola in farmland soil.

Cadmium (Cd) pollution in farmland soils severely affects agricultural production safety, thereby threatening human health. Sedum plumbizincicola is a Cd and Zn hyperaccumulator commonly used for the phytoremediation of Cd-contaminated soil. This study was aimed to improve the remediation effect of S. plumbizincicola on Cd-contaminated farmland soil and provide a theoretical basis for the enhancement of endophytic bacteria in the repair of Cd-contaminated soil with S. plumbizincicola. Four kinds of endophytic bacteria, namely Buttiauxella, Pedobacter, Aeromonas eucrenophila, and Ralstonia pickettii, were used, and soil culture experiments and pot experiments were conducted to explore the effects of endophytic bacteria on soil Cd speciation and phytoremediation efficiency of Cd-contaminated farmland soils. Under the experimental conditions, after inoculation with endophytic bacteria, the soil pH was effectively reduced, content of weak acid-extracted Cd and oxidizable Cd increased, and content of reducible Cd and residual Cd decreased. Soil Cd activity was increased, and the availability coefficient of soil Cd increased by 1.15 to 6.41 units compared with that of the control (CK2). Compared with CK2, the biomass of S. plumbizincicola significantly increased by 23.23-55.12%; Cd content in shoots and roots of S. plumbizincicola increased by 29.63-46.01% and 11.42-84.47%, respectively; and bioconcentration factor was 2.13 to 2.72 times that of CK2. The Cd removal rate of S. plumbizincicola monocropping was 48.25%. When S. plumbizincicola was planted with inoculating endophytic bacteria, the Cd removal rate in the soil reached 61.18-71.49%, which was significantly higher than that of CK2 (p < 0.05). The treatment with endophytic bacteria activated soil Cd, promoted the growth of S. plumbizincicola, increased its Cd content, and enhanced the phytoremediation of Cd-contaminated farmland soil. Therefore, endophytic bacteria can be used to improve the remediation efficiency of S. plumbizincicola in Cd-contaminated farmland soils.