Phylogenomics and plastomics offer new evolutionary perspectives on Kalanchoideae (Crassulaceae).

BACKGROUND AND AIMS: Kalanchoideae is one of three subfamilies within Crassulaceae and contains four genera. Despite previous efforts, the phylogeny of Kalanchoideae remains inadequately resolved with persistent issues including low support, unstructured topologies and polytomies. This study aimed to address two central objectives: (1) resolving the pending phylogenetic questions within Kalanchoideae by using organelle-scale 'barcodes' (plastomes) and nuclear data; and (2) investigating interspecific diversity patterns among Kalanchoideae plastomes. METHODS: To explore the plastome evolution in Kalanchoideae, we newly sequenced 38 plastomes representing all four constituent genera (Adromischus, Cotyledon, Kalanchoe and Tylecodon). We performed comparative analyses of plastomic features, including GC and gene contents, gene distributions at the IR (inverted repeat) boundaries, nucleotide divergence, plastomic tRNA (pttRNA) structures and codon aversions. Additionally, phylogenetic inferences were inferred using both the plastomic dataset (79 genes) and nuclear dataset (1054 genes). KEY RESULTS: Significant heterogeneities were observed in plastome lengths among Kalanchoideae, strongly correlated with LSC (large single copy) lengths. Informative diversities existed in the gene content at SSC/IRa (small single copy/inverted repeat a), with unique patterns individually identified in Adromischus leucophyllus and one major Kalanchoe clade. The ycf1 gene was assessed as a shared hypervariable region among all four genera, containing nine lineage-specific indels. Three pttRNAs exhibited unique structures specific to Kalanchoideae and the genera Adromischus and Kalanchoe. Moreover, 24 coding sequences revealed a total of 41 lineage-specific unused codons across all four constituent genera. The phyloplastomic inferences clearly depicted internal branching patterns in Kalanchoideae. Most notably, by both plastid- and nuclear-based phylogenies, our research offers the first evidence that Kalanchoe section Eukalanchoe is not monophyletic. CONCLUSIONS: This study conducted comprehensive analyses on 38 newly reported Kalanchoideae plastomes. Importantly, our results not only reconstructed well-resolved phylogenies within Kalanchoideae, but also identified highly informative unique markers at the subfamily, genus and species levels. These findings significantly enhance our understanding of the evolutionary history of Kalanchoideae.

Root-associated bacterial microbiome shaped by root selective effects benefits phytostabilization by Athyrium wardii (Hook.).

The root-associated microbiome assembly substantially promotes (hyper)accumulator plant growth and metal accumulation and is influenced by multiple factors, especially host species and environmental stress. Athyrium wardii (Hook.) is a phytostabilizer that grows in lead (Pb)-zinc (Zn) mine tailings and shows high root Pb accumulation. However, there remains little information on the assembly of the root-associated microbiome of A. wardii and its role in phytostabilization. A field study investigated the structural and functional variation in the root-associated bacterial microbiome of Athyrium wardii (Hook.) exposed to different levels of contamination in Pb-Zn mine tailings. The root compartment dominated the variation in the root-associated bacterial microbiome but the levels of contaminants showed less impact. Bacterial co-occurrence was enhanced in the rhizosphere soil and rhizoplane but tended to be much simpler in the endosphere in terms of network complexity and connectivity. This indicates that the microbial community assembly of A. wardii was non-random and shaped by root selective effects. Proteobacteria, Chloroflexi, Actinobacteria, Cyanobacteria, and Acidobacteriota were generally the dominant bacterial phyla. The genera Crossiella and Bradyrhizobium were enriched in the rhizosphere and cyanobacterial genera were enriched in the endosphere, demonstrating substantial advantages to plant survival and adaptation in the harsh mine environment. Functional categories involved in amino acid and carbohydrate metabolism were abundant in the rhizosphere soil, thus contributing to metal solubility and bioavailability in the rhizosphere. Membrane transporters, especially ATP-binding cassette transporters, were enriched in the endosphere, indicating a potential role in metal tolerance and transportation in A. wardii. The study shows substantial variation in the structure and function of microbiomes colonizing different compartments, with the rhizosphere and endophytic microbiota potentially involved in plant metal tolerance and accumulation during phytostabilization.

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.

Optimizing Soil Sampling for Accurately Prediction of the Potential Remediation-Effective Area in a Contaminated Agricultural Land.

To achieve food security in a contaminated agricultural land, the remediation areas usually need more samples to obtain accurate contamination information and implement appropriate measures. In this study, we propose an optimal encryption sampling design to instead of the detailed survey, which is determined by the variation of heavy metals and the technology capability of remediation, to guide soil sampling for accurately remediation in the potential remediation-effective areas (PRA). The coefficient of screening variation threshold (CSVT), considering spatial variation, technology capacity and acceptable error of sampling, together with the spatial cyclic statistics method of neighbourhood analysis, is introduced to identify and delineate the PRA. Both of the hypothetical analysis and application case studies are conducted to illustrate the advantages and disadvantages of the optimization. The results show that, compared with the detailed survey, the optimal design shows a lower overall accuracy due to its sparsely sampling at the clean area, but it exhibits a similar effect of accurately prediction in boundary delineation and further classification in the PRA in both simulation and application studies. This work provides an effective method for subsequent accurate remediation at the investigation stage and valuable insights into application combination of technology capacity and contaminated agricultural land investigation.

Mobilization of Colloid- and Nanoparticle-Bound Arsenic in Contaminated Paddy Soils during Reduction and Reoxidation.

The association of arsenic (As) with colloidal particles could facilitate its transport to adjacent water systems or alter its availability in soil-rice systems. However, little is known about the size distribution and composition of particle-bound As in paddy soils, particularly under changing redox conditions. Here, we incubated four As-contaminated paddy soils with distinctive geochemical properties to study the mobilization of particle-bound As during soil reduction and subsequent reoxidation. Using transmission electron microscopy-energy dispersive spectroscopy and asymmetric flow field-flow fractionation, we identified organic matter (OM)-stabilized colloidal Fe, most likely in the form of (oxy)hydroxide-clay composite, as the main arsenic carriers. Specifically, colloidal As was mainly associated with two size fractions of 0.3-40 and >130 kDa. Soil reduction facilitated the release of As from both fractions, whereas reoxidation caused their rapid sedimentation, coinciding with solution Fe variations. Further quantitative analysis demonstrated that As concentrations positively correlated with both Fe and OM concentrations at nanometric scales (0.3-40 kDa) in all studied soils during reduction and reoxidation, yet the correlations are pH-dependent. This study provides a quantitative and size-resolved understanding of particle-bound As in paddy soils, highlighting the importance of nanometric Fe-OM-As interactions in paddy As geochemical cycling.

Zinc Supply Affects Cadmium Uptake and Translocation in the Hyperaccumulator Sedum Plumbizincicola as Evidenced by Isotope Fractionation.

This study employs stable isotope analysis to investigate the mechanisms of cadmium (Cd) and zinc (Zn) interaction in the metal hyperaccumulating plant species Sedum plumbizincicola. To this end, the Cd and Zn isotope compositions of root, stem, leaf, and xylem sap samples were determined during metal uptake and translocation at different Cd and Zn concentrations. The enrichment of light isotopes of both elements in plants during uptake was less pronounced at low metal supply levels, likely reflecting the switch from a low-affinity to a high-affinity transport system at lower levels of external metal supply. The lower δ114/110Cd values of xylem sap when treated with a metabolic inhibitor decreasing the active Cd uptake further supports the preference of heavier Cd isotopes during high-affinity transport. The Δ66Znplant-initial solution or Δ66Znplant-final solution values were similar at different Cd concentrations, indicating negligible interaction of Cd in the Zn uptake process. However, decreasing Zn supply levels significantly increased the enrichment of light Cd isotopes in plants (Δ114/110Cd = -0.08‰) in low-Cd treatments but reduced the enrichment of light Cd isotopes in plants (Δ114/110Cd = 0.08‰) under high Cd conditions. A systematic enrichment of heavy Cd and light Zn isotopes was found in root-to-shoot translocation of the metals. The Cd concentrations of the growth solutions thereby had no significant impact on Zn isotope fractionation during root-to-shoot translocation. However, the Δ114/110Cdtranslocation values hint at possible competition between Cd and Zn for transporters during root-to-shoot transfer and this may impact the transport pathway of Cd. The stable isotope data demonstrate that the interactions between the two metals influenced the uptake and transport mechanisms of Cd in S. plumbizincicola but had little effect on those of Zn.

Plastome evolution of Aeonium and Monanthes (Crassulaceae): insights into the variation of plastomic tRNAs, and the patterns of codon usage and aversion.

MAIN CONCLUSION: This study reported 13 new plastomes from Aeonium and Monanthes, and observed new markers for phylogeny and DNA barcoding, such as novel tRNA structures and codon usage bias and aversion. The Macaronesian clade of Crassulaceae consists of three genera: Aichryson, with about 15 species; Monanthes, with about 10 species; Aeonium, with about 40 species. Within this clade, Aeonium, known as "the botanical equivalent of Darwin's finches", is regarded as an excellent model plant for researching adaptive evolution. Differing from the well-resolved relationships among three genera of the Macaronesian clade, the internal branching patterns within the genus Aeonium are largely unclear. In this study, we first reported 13 new plastomes from genus Aeonium and the closely related genus Monanthes. We further performed comprehensive analyses of the plastomes, with focuses on the secondary structures of pttRNAs and the patterns of codon usage and aversion. With a typical circular and quadripartite structure, the 13 plastomes ranged from 149,900 to 151,030 bp in size, and the unique pattern in IR junctions might become a family-specific marker for Crassulaceae species. Surprisingly, the π values of plastomes from Monanthes were almost twice those from Aeonium. Most importantly, we strongly recommend that highly polymorphic regions, novel putative pttRNA structures, patterns of codon usage bias and aversion derived from plastomes might have phylogenetic implications, and could act as new markers for DNA barcoding of plants. The results of phylogenetic analyses strongly supported a clear internal branching pattern in Macaronesian clade (represented by Aeonium and Monanthes), with higher nodal support values. The findings reported here will provide new insights into the variation of pttRNAs, and the patterns of codon usage and aversion of the family Crassulaceae.

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.

Potential mobilization of cadmium and zinc in soils spiked with smithsonite and sphalerite under different water management regimes.

Particulate cadmium (Cd) and zinc (Zn) are ubiquitous in agricultural soils of Pb-Zn mining regions. Water management serves as an important agronomic measure altering the bioavailability of Zn and Cd in soils, but how this affects particulate Cd and Zn and the underlying mechanisms remain largely unknown. Microcosm soil incubation combined with spectroscopic and microscopic characterization was conducted. During a two-year-long incubation period we observed that the concentrations of soil CaCl2-extractable Zn and Cd increased 3-10 times in sphalerite-spiked soils and 1-2 times in smithsonite-spiked soils under periodic flooding conditions due to the long-term dissolution of sphalerite (SP) and smithsonite (SM). However, the increase in the concentration of CaCl2-extractable metals (Zn: from 0.607 mg kg-1 to 1.051 mg kg-1 and Cd: from 0.047 mg kg-1 to 0.119 mg kg-1) was found only in SP-treatment under continuous flooding conditions, indicating the mobilization of metals. Ultrafiltration analysis shows that the nanoparticulate fraction of Zn and Cd in soil pore water increased 5 and 7 times in SP-treatments under continuous flooding conditions, suggesting the increment of metal pools in soil pore water. HRTEM-EDX-SAED further reveals that these nanoparticles were mainly crystalline ZnS and Zn-bearing sulfate nanoparticles in the SP-treatment and amorphous ZnCO3 and ZnS nanoparticles in the SM-treatment. Therefore, the formation of the stable crystalline Zn-bearing nanoparticles in the SP-treatment may explain the elevation of the concentration of soil CaCl2-extractable Zn and Cd under continuous flooding. The potential mobility of particulate metals should therefore be expected in scenarios of continuous flooding such as paddy soils and wetland systems.

Culture salinity modulates Nrf2 antioxidant signaling pathway and immune response of juvenile Genetically Improved Farmed Tilapia (GIFT) (Oreochromis niloticus) under different dietary protein levels.

This study aimed to evaluate that dietary protein levels and culture salinity levels affect the health status of juvenile genetically improved farmed tilapia (GIFT, Oreochromis niloticus). Graded protein levels of six diets were prepared, ranging from 18.20% to 49.49% (dry basis), and were used in cultured GIFT at two salinity levels (0‰ and 8‰) for 8 weeks. The results suggested that appropriate protein levels reduced pro-inflammatory gene expressions in the intestine including interleukin 1ß (IL-1ß), interleukin 8 (IL-8) and tumour necrosis factor-α (TNF-α) mRNA levels at two salinity levels (P < 0.05). 8‰ salinity significantly decreased the expression levels of IL-1ß, TNF-α and nuclear factor-kappa B (NF-κB) (P < 0.05). The anti-inflammatory factor interleukin 10 (IL-10) was significantly increased by 36.42% protein level (P < 0.05). Regarding antioxidant capacity, appropriate protein levels and 8‰ salinity significantly improved the antioxidant capacity of fish by regulating the activities of intestinal total superoxide dismutase (T-SOD), glutathione peroxidase (GPx), and the levels of glutathione (GSH) and malondialdehyde (MDA). Furthermore, appropriate protein levels and 8‰ salinity also significantly enhanced the antioxidant gene expressions associated with the Nrf2/keap1 signaling pathway by regulating the expression levels of heme oxygenase-1 (HO-1), GPx, catalase (CAT) and superoxide dismutase (SOD). According to GPx activities and the mRNA levels of IL-10, the optimum dietary protein levels for GIFT juveniles were 31.12%-32.18% (0‰) and 34.25-35.38% (8‰) based on second-degree polynomial regression analysis. The present study found that appropriate protein levels and 8‰ culture salinity are critical in maintaining the health of GIFT juveniles by improving antioxidant and immune capacity.

Integrated Life Cycle Assessment for Sustainable Remediation of Contaminated Agricultural Soil in China.

Agricultural land degradation is posing a serious threat to global food security. Restoration of the degraded land has traditionally been viewed as an inherently sustainable practice; however, restoration processes render consequential environmental impacts which could potentially exceed the benefit of restoration itself. In the present study, an integrated life cycle assessment analysis was conducted to evaluate life cycle primary, secondary, and tertiary impacts associated with the restoration of the contaminated agricultural land. The results demonstrated the importance of including spatially differentiated impacts associated with managing the land and growing crops. Comparing four risk management scenarios at a contaminated field in Southern China, it was found that the primary and secondary impacts followed the order of no action > chemical stabilization > phytoextraction > alternative planting. However, when tertiary impacts were taken into account, alternative planting rendered much higher footprint in comparison with phytoextraction and chemical stabilization, which provides evidence against an emerging notion held by some policy makers. Furthermore, assuming that the loss of the rice paddy field in Southern China is compensated by the deforested land in the Amazon rainforest, the total global environmental impact would far exceed that of no action, resulting in 687 ton CO2-e ha-1 of climate change impact. Overall, the present study provides new research findings to support more holistic policy making and also sheds lights on the future development of various restoration technologies.

Ecotoxicity of arsenic contamination toward the soil enchytraeid Enchytraeus crypticus at different biological levels: Laboratory studies.

The ecotoxicity of arsenic (As) contamination toward small soil fauna living in soil pore water such as soil enchytraeids has rarely been studied but is important in the assessment of soil pollution. Here, the endpoints of As ecotoxicity to Enchytraeus crypticus were studied at three biological levels, i.e., individual (morphology and body tissue As concentrations), population (survival, reproduction and growth) and cell biochemistry (antioxidant enzymes CAT, POD and SOD and peroxidation malondialdehyde MDA). Contact filter paper tests without soil and single species tests with OECD artificial and field soils were conducted. Arsenic contamination resulted in severe morphological pathologies in E. crypticus and the symptoms and degree of damage increased gradually with increasing As concentration and exposure time up to 48 h. The abnormal morphological effects occurred before the impairment of fecundity. The population endpoints responded to the As concentration and the EC50 values increased in the following sequence: reproduction, juvenile body weight, adult body weight, juvenile length and adult length. Changes in biochemistry parameters were induced rapidly and changed with increasing As concentration and exposure time. The activity peak values of enzymes were 3-5 times higher and the activity maximum values of MDA were 1-3 time higher than their controls. The sensitivity of enzyme activities was generally much higher than that of MDA and CAT generally showed the highest enzyme activity. The results indicate that As contamination can be very harmful to soil enchytraeids and the endpoints of the ecotoxicity tests of soil enchytraeids can be used to complement existing soil As assessment systems or may be used alone for the assessment of soil As pollution.

Remediation of a metal-contaminated soil by chemical washing and repeated phytoextraction: a field experiment.

Agricultural soil contaminated with potentially toxic metals poses great health risk to humans and it requires long-term remediation. Here, we investigate the remediation of metal-polluted agricultural soil by combining chemical washing with repeated phytoextraction. The polluted field was initially washed with 40 mmol L-1 FeCl3 (F) or 20 mmol L-1 FeCl3 + 40 mmol L-1 citric acid (F + C). After the application of organic fertilizer (O), lime (L), and sepiolite (S), Sedum plumbizincicola was cultivated for three successive crops from 2017 to 2019. Results showed that the soil washed with FeCl3 had high removal efficiencies of Cd (35.2%), Pb (24.3%), and Zn (26.6%). Although the shoot biomass and metal concentrations of S. plumbizincicola decreased significantly in the first crop, there were no significant differences in the subsequent two crops. Throughout the remediation process, the higher total removal efficiencies of Cd, Pb, and Zn were conducted in F + OLS treatment which observed in 71.0, 34.0, and 47.7%, respectively. The results, therefore, conclusively indicated that combining chemical washing with repeated phytoextraction showed considerable potential for the remediation of agricultural soils polluted with multiple metals. However, further studies are required to focus on the amelioration of the degraded soil quality and safe agricultural production.

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.

Auxin-Induced SaARF4 Downregulates SaACO4 to Inhibit Lateral Root Formation in Sedum alfredii Hance.

Lateral root (LR) formation promotes plant resistance, whereas high-level ethylene induced by abiotic stress will inhibit LR emergence. Considering that local auxin accumulation is a precondition for LR generation, auxin-induced genes inhibiting ethylene synthesis may thus be important for LR development. Here, we found that auxin response factor 4 (SaARF4) in Sedum alfredii Hance could be induced by auxin. The overexpression of SaARF4 decreased the LR number and reduced the vessel diameters. Meanwhile, the auxin distribution mode was altered in the root tips and PIN expression was also decreased in the overexpressed lines compared with the wild-type (WT) plants. The overexpression of SaARF4 could reduce ethylene synthesis, and thus, the repression of ethylene production decreased the LR number of WT and reduced PIN expression in the roots. Furthermore, the quantitative real-time PCR, chromatin immunoprecipitation sequencing, yeast one-hybrid, and dual-luciferase assay results showed that SaARF4 could bind the promoter of 1-aminocyclopropane-1-carboxylate oxidase 4 (SaACO4), associated with ethylene biosynthesis, and could downregulate its expression. Therefore, we concluded that SaARF4 induced by auxin can inhibit ethylene biosynthesis by repressing SaACO4 expression, and this process may affect auxin transport to delay LR development.

Cadmium and Lead Pollution Characteristics of Soils, Vegetables and Human Hair Around an Open-cast Lead-zinc Mine.

Atmospheric deposition of cadmium (Cd) and lead (Pb) was investigated together with the accumulation, distribution and health risks from potentially toxic metals in soils, vegetables and human hair at a mining area in southwest China. Annual atmospheric deposition of Cd and Pb were 41.1 and 192 g ha- 1, respectively, and consisted mainly of dry deposition. Agricultural soils experienced high levels of metal pollution around the mine, with 66.4% and 57.3 % of vegetable samples grown on these polluted fields exceeding maximum permissible Cd and Pb concentrations, particularly the leafy vegetables. Residents living near the mining area had high Cd (0.75 mg kg- 1) and Pb (6.87 mg kg- 1) concentrations in their hair, and the maximum values occurred in occupationally exposed individuals. Long-term mining activities have resulted in high health risks to the local population due to Cd and Pb deposition and accumulation from the atmosphere, soils and vegetables.

Soil Metal Immobilization in Agricultural Land Contaminated with Cadmium and Lead: A Case Study of Effectiveness Evaluation in Lanping, Southwest China.

The growth of edible crops on land that is highly polluted with potentially toxic elements is prohibited in many developed countries, but the growth of fiber or energy crops may be permitted. Here, we have evaluated metal immobilization in a maize field polluted with cadmium (Cd) and lead (Pb) to determine the thresholds of soil CaCl2-extractable Cd and Pb and to assess management options designed to maximize food safety. Based on geographical and statistical methods we found that when the soil pH was increased from 5.24 to 6.24, the soil CaCl2-extractable Cd and Pb values decreased by 47.8 and 74.7%, respectively. Soil CaCl2-extractable Pb concentrations need to be < 2.14 mg kg-1 in order to comply with the Chinese maximum permissible grain Pb concentration (< 0.2 mg kg-1). Immobilization increased the percentage of samples that were below permissible levels from 77.4% to 96.2% (grain Cd) and 90.6% to 96.2% (grain Pb) during the period 2017 to 2019. To avoid excessive or inadequacy immobilization, the spatial distribution of correlation coefficients of soil pH, CaCl2-extractable or grain Cd/Pb may be helpful in the precise management of immobilization for long-term remediation.

Estimation of Heavy Metals in Tailings and Soils Using Hyperspectral Technology: A Case Study in a Tin-Polymetallic Mining Area.

Rapid assessment of heavy metal (HM) pollution in mining areas is urgently required for further remediation. Here, hyperspectral technology was used to predict HM contents of multi-media environments (tailings, surrounding soils and agricultural soils) in a mining area. The correlation between hyperspectral data and HMs was explored, then the prediction models were established by partial least squares regression (PLSR) and back propagation neural networks (BPNN). The determination coefficients (R2), root mean squared error and ratios of performance to interquartile range (RPIQ) were used to evaluate the performance of the models. Results show that: (1) both PLSR and BPNN had good prediction ability, and (2) BPNN had better generalization ability (Cu (R2 = 0.89, RPIQ = 3.05), Sn (R2 = 0.86, RPIQ = 4.91), Zn (R2 = 0.74, RPIQ = 1.44) and Pb (R2 = 0.70, RPIQ = 2.10)). In summary, this study indicates that hyperspectral technology has potential application in HM estimation and soil pollution investigation in polymetallic mining areas.

Pollution Characteristics and Risk Assessment of Potential Toxic Elements in a Tin-polymetallic Mine Area Southwest China: Environmental Implications by Multi-Medium Analysis.

A multi-medium system, involving tailing area (tailings, surrounding soils and water) and downstream agricultural area (river water, sediments and farmland soils), was conceived to evaluate the pollution status of potential toxic elements (PTEs, including Fe, Mn, Ni, Cu, Zn, As, Sn, Pb, Cr and Cd) and environmental risks in a tin-polymetallic mine area southwest China. The results indicated that tailings exhibited representative enrichment and combination characteristics of Sn, Cu, Ni, Fe, As, Pb and Cr compared to surrounding soils. Acid mine drainage (AMD) from tailings and other mining-related sources greatly affected river water and farmland soils, resulting in soil acidification and accumulation of Sn, As, Cu and Pb in paddy soils. Overall, potential ecological risks posed by tailings and river sediments, and pollution risks from Cu, As and Pb in farmland should be concerned. Therefore, effective measures should be urgently taken to prevent PTEs and AMD into surrounding environmental media.