[Two-stage Inhibition Effects of Burkholderia sp. Y4 Application on Cadmium Uptake and Transport in Wheat].

In order to evaluate the feasibility of using Burkholderia sp. Y4 as a cadmium （Cd）-reducing bacterial agent in contaminated wheat fields， the changes in the rhizosphere soil microbial community and Cd available state， as well as the content and transport characteristics of Cd in the wheat root， basal node， internode， and grain under the treatment of strain Y4 were tested using microbial high-throughput sequencing， step-by-step extraction， subcellular distribution， and occurrence analyses. The results showed that root application of strain Y4 significantly reduced the root and grain Cd content of wheat by 7.7% and 30.3%， respectively， compared with that in the control treatment. The Cd content and Cd transfer factor results in wheat vegetative organs showed that strain Y4 reduced the Cd transfer factor from basal node to internode by 79.3%， and Cd content in the wheat internode stem also decreased by 50.9%. The study of Cd occurrence morphology showed that strain Y4 treatment increased the proportion of residual Cd in roots and basal ganglia， decreased the contents of inorganic and water-soluble Cd in roots， and increased the content of residual Cd in basal ganglia. Further examination of the subcellular distribution of Cd showed that the Cd content in root cell walls and basal ganglia cell fluid increased by 21.3% and 98.2%， respectively， indicating that the Cd fixation ability of root cell walls and basal ganglia cell fluid was improved by the strain Y4 treatment. In the rhizosphere soil， it was found that the microbial community structure was changed by strain Y4 application. Under the Y4 treatment， the relative abundance of Burkholderia increased from 9.6% to 11.5%， whereas that of Acidobacteriota decreased. Additionally， the relative abundance of Gemmatimonadales， Pseudomonadales， and Chitinophagales were also increased by strain Y4 treatment. At the same time， the application of strain Y4 increased the pH value of rhizosphere soil by 8.3%. The contents of exchangeable Cd， carbonate-bound Cd， and iron-manganese oxide-bound Cd in the soil decreased by 44.4%， 21.7%， and 15.9%， respectively， whereas the proportion of residual Cd reached 53.6%. Root application of strain Y4 increased the contents of nitrate nitrogen and ammonium nitrogen in the soil by 22.0% and 21.4%， respectively， and the contents of alkaline nitrogen also increased to a certain extent. In conclusion， the root application of strain Y4 not only improved soil nitrogen availability but also inhibited Cd transport and accumulation from contaminated soil to wheat grains in a "two-stage" manner by reducing Cd availability in rhizosphere soil and improving Cd interception and fixation capacity of wheat roots and basal nodes. Therefore， Burkholderia Y4 has application potential as a Cd-reducing and growth-promoting agent in wheat.

[Effects of Burkholderia sp. Y4 on Cadmium Damage and Uptake in Rice Seedlings].

In order to investigate the effects of Burkholderia sp. Y4 on rice seedlings under cadmium (Cd) stress, seed germination and vermiculite culture experiments were conducted using low Cd-accumulation xiangzaoxian 24 (X24) and high Cd-accumulation Tyou 705 (T705) varieties. The effects of Burkholderia sp. Y4 on rice growth, oxidative damage caused by Cd, and Cd accumulation were studied. Additionally, the Cd2+ flux rates in the elongation zone of rice roots under Burkholderia sp. Y4 application were detected using non-invasive micro-test technology. Burkholderia sp. Y4 alleviated the inhibition effect of Cd on rice seed germination by 13.8%. After inoculation with Burkholderia sp. Y4 for 7 d, the length of rice roots and buds increased by 83.3% and 12.2%, and their dry weight increased by 56.8% and 12.5%, respectively; those in the 10 d Y4 inoculation group increased by 28.6% and 20.0% in length and by 113.2% and 46.0% in dry weight, respectively. Burkholderia sp. Y4 inoculation also alleviated rice oxidative stress damage caused by Cd. The application of strain Y4 significantly reduced the content of the oxidative damage product malondialdehyde (MDA) in the shoots and roots of rice seedlings by 21.5% and 16.9%, respectively. Under Burkholderia sp. Y4 inoculation, the significant changes in antioxidant enzyme SOD and CAT activities caused by Cd stress disappeared in rice roots; those in shoots also decreased from 176.9% and 74.8% to 53.3% and 21.5%, respectively. Conversely, Burkholderia sp. Y4 inhibited Cd uptake by rice seedlings with different genotypes, including the low Cd-accumulation variety X24 and high Cd-accumulation variety T705. The root application of strain Y4 significantly reduced Cd accumulation in the shoots and roots of rice seedlings by 79.2% and 62.7% in T705 and by 57.3% and 24.1% in X24, respectively. The Cd2+ flux rate of high Cd-accumulation variety T705 was significantly higher than that of low Cd-accumulation variety X24. Under Burkholderia sp. Y4 inoculation, the yellow membrane was formed on the root surface of rice seedlings, and the Cd2+ flux rate in the elongation zone of T705 and X24 roots decreased by 36.0% and 35.0% in 3-day-old seedlings, as well as by 44.6% and 24.9% in 10-day-old seedlings, respectively. In conclusion, Burkholderia sp. Y4 inoculation inhibited the toxic effects of Cd on rice seedling growth through alleviating oxidative stress and damage caused by Cd. Furthermore, the root application of Burkholderia sp. Y4 effectively decreased the Cd2+ flux rate in the elongation zone of roots to inhibit the Cd uptake and accumulation in roots and shoots of rice seedlings. This study provides theoretical basis and data support for the application of Burkholderia sp. Y4 as a Cd-reducing and growth-promoting agent for rice in contaminated farmland.

[Isolation and Identification of the Plant Endophyte R-13 and Its Effect on Cadmium Accumulation in Solanum nigrum L.]

The combination of endophytes and hyperaccumulator plants can significantly improve the efficiency of heavy metal phytoremediation in contaminated soil. A plant endophyte named Herbaspirillum R-13 was isolated from rice roots in a cadmium (Cd) contaminated paddy field. This strain exhibited a strong tolerance to Cd2+ and could grow on a solid medium with a Cd2+ concentration of 300 mg·kg-1. The R-13 strain was able to produce siderophores and Indole acetic acid (IAA), through color reactions. In addition, Pikovskaya's and Ashby's solid medium tests showed that the R-13 strain had a lower capacity for dissolving phosphorus but a higher capacity for fixing nitrogen. In the pot experiment, high-throughput sequencing technology was used to track the colonization of the R-13 strain in Solanum nigrum L. roots. Three days after inoculation, the relative abundance of Herbaspirillum in the root of Solanum nigrum L. had increased by 201.88% compared to the blank control (CK) and after two inoculations, the relative abundance of Herbaspirillum in the root of Solanum nigrum L. had increased by 1182.44% compared to CK. The relative abundance of Herbaspirillum in the root of Solanum nigrum L. began to decrease significantly from 5 days after inoculation. Inoculation with 20 mL·pot-1 of R-13 fermentation broth resulted in no significant effects on the Cd content of roots, stems, leaves, or fruits of S. nigrum L. With 40 mL·pot-1 of fermentation broth, the Cd content of vegetative organs and fruits was significantly increased. When it reached 200 mL·pot-1, the Cd content of vegetative organs was the highest, with Cd concentrations in the roots, stems, leaves, and fruits increasing by 84.42%, 43.67%, 64.06%, and 20.29%, respectively. In conclusion, root inoculation with endophytic Herbaspirillum R-13 can significantly increase the relative content of Herbaspirillum in the root system and enhance Cd absorption of S. nigrum L. Therefore, this strain has excellent prospects for application in the phytoremediation of soil contaminated with Cd.

[Foliar Application of L-Cysteine: Effects on the Concentration of Cd and Mineral Elements in Rice].

To evaluate the feasibility of applying L-cysteine (L-Cys) as a foliar conditioner for Cd reduction in rice, a field experiment was conducted to investigate the concentration of Cd and mineral elements in rice after the foliar application of L-Cys. The variation in Cd distribution and morphology in key rice organs was examined to study the Cd reduction mechanisms of spraying L-Cys on rice. The results showed that foliar application of L-Cys at the rice-flowering stage significantly decreased Cd concentration in grains, in a concentration dependent manner, without inhibiting the accumulation of mineral elements Ca, Mg, K, Mn, and Zn. With a 10 mmol ·L-1 L-Cys application, Cd concentration in rice grains decreased by 59.2%, to below 0.2 mg ·kg-1, which is the maximum safety limit in China. Foliar application of L-Cys also inhibited Cd accumulation in rice vegetative organs, including rachises, first nodes, neck-panicles, flag leaves, second internodes, second nodes, second leaves, stalks, and roots (58.3%, 56.0%, 62.7%, 67.0%, 59.3%, 61.5%, 60.2%, 54.9%, and 50.3%, respectively). After transfer factor calculation, first nodes were found to be the key organ for Cd blocking in rice. The application of L-Cys increased Cd transfer from flag leaves and second internodes to first nodes (105.4% and 45.8%, respectively), but decreased Cd transfer from first nodes up to neck-panicles (27.5%). In rice first nodes, the concentrations of Cd in the inorganic, water soluble, and residue states were all lower following L-Cys application, and the proportion of residual Cd increased to 94.4%. Therefore, foliar application of L-Cys significantly inhibited Cd transport and accumulation in rice grains, by decreasing the Cd concentrations of various vegetative organs and improving Cd interception in the first nodes. This is a promising way to produce rice with lower Cd concentrations and normal mineral element concentrations in Cd-contaminated paddy fields.

[Mechanism of S-allyl-L-cysteine Alleviating Cadmium Stress in Seedling Roots and Buds of Rice Seedlings].

Cd has toxic effects on rice seed germination and plant growth, which may eventually lead to decreased yield and excessive Cd content in rice grains. The potential mechanism of S-allyl-L-cysteine (SAC), a natural sulfur compound derived from garlic extract, in alleviating Cd2+ stress in young roots and buds of rice seedlings was studied by a seed germination experiment. "Zhong zao 35", one of the main rice varieties in Southern China, was selected as the test material. Firstly, the alleviating effect of SAC on Cd2+ stress in rice seedling roots and buds was studied. Following this, the physiological mechanism of Cd2+ stress alleviation by SAC was examined based on the expression of the Cd transporter coding gene using real-time fluorescent quantitative PCR. The results showed that when the Cd2+ stress concentration reached 50 µmol·L-1, the young roots and buds of rice seedlings were significantly inhibited, and when the SAC concentration reached 200 µmol·L-1, Cd2+ stress was significantly alleviated. Compared to a Cd2+ stress treatment group, the total root length, surface area, and volume of young roots was increased by 173.5%, 65.52%, and 37.04%, respectively; CAT and SOD activity in young roots and buds was increased by 212.42% and 110.76%, and 31.41% and 47.31%, respectively; MDA and GSH content was decreased by 43.09% and 34.12%, and 33.97% and 35.74%, respectively; and Cd content was decreased by 35.91% and 28.86%, respectively. The results of quantitative real-time PCR showed that the relative expression levels of OsNramp5 and OsHMA2 were significantly reduced by 33.38% and 34.99% compared with the Cd2+ stress group, respectively. However, the relative expression level of OsHMA3 was significantly increased by 33.96%. From the above experimental results, the main mechanism by which SAC reduces Cd2+ stress in the young roots and buds of rice is via the regulation of Cd transporter-encoding genes, reducing Cd2+ transport to young roots and buds, and increasing transport to vacuoles.

[Effects of Foliar Spraying of 2,3-dimercaptosuccinic Acid on Cadmium Uptake, Transport, and Antioxidant System in Rice Seedlings].

Rice contaminated by Cd has aroused widespread public concern. It is of great importance to find effective ways to reduce Cd translocation from roots to shoots and alleviate Cd stress in rice to ensuring food quality and safety. In this study, 2,3-dimercaptosuccinic acid (DMSA) was sprayed onto the leaves of rice seedlings to evaluate the feasibility of DMSA reducing Cd translocation to rice shoots and alleviating Cd stress. Therefore, seedlings of Zhongzao 35, one of the main rice cultivars in southern China, were used to study the effects of different concentrations of DMSA on the uptake and transport of Cd in rice seedlings by hydroponics. The effects of DMSA on MDA and GSH content, and activities of antioxidant enzymes such as CAT and SOD in rice seedlings, were also investigated. The results showed that after four iterations of foliar application of DMSA at concentrations of 0.2, 0.4, and 1.0 mmol·L-1, the Cd concentration in the rice seedling shoots decreased significantly with increasing DMSA spraying concentration. Compared with the control, the Cd concentration in shoots decreased by 22.1%, 39.7%, and 43.5%, respectively, but had no significant effect on the root Cd concentration. There was no significant effect on the concentrations of K, Ca, Mg, Fe, Zn, or Mn in the shoot or root. The content of MDA and GSH in the shoots of rice seedlings decreased significantly after four spraying times of DMSA and the activity of CAT and SOD increased significantly, which shows that spraying DMSA alleviated the stress effect of Cd on rice seedlings. Foliar application of DMSA can significantly reduce the accumulation of Cd in rice shoots but has no significant effect on the content of six common mineral elements, and can effectively relieve the oxidative damage caused by Cd stress. DMSA has the potential to develop a foliar modulator for reducing rice grain Cd content.

Rice vegetative organs alleviate cadmium toxicity by altering the chemical forms of cadmium and increasing the ratio of calcium to manganese.

Altering Cd chemical form is one of the mechanisms to alleviate Cd toxicity in rice plant. Field experiments were carried out in this study to investigate the potential of rice vegetative organs in altering Cd into insoluble chemical forms in the natural environment. Experimental results showed that more than 80% of Cd in rice roots existed in the insoluble forms. Uppermost nodes altered Cd into insoluble form preferentially and generally had higher content of insoluble Cd than other organs. Rachises displayed a slow increasing trend in soluble Cd when total Cd in roots was less than 1.8 mg kg-1. However, when Cd content in roots exceeded 2.8 mg kg-1, the ratio of insoluble to soluble Cd remained stable at 85:15 in rachises and roots, and at 75:25 in uppermost nodes and flag leaves. Cd concentration in grains was greatly lower than that in vegetative organs, and closely correlated with the content of soluble Cd in rachises (r = 0.991**) as well as in uppermost nodes. Soluble Cd in the uppermost nodes displayed a much lower mobility than that in other organs. Accumulation of soluble Cd was always companied by decrease of Ca and increase of Mn in roots, uppermost nodes and rachises. A small increase of soluble Cd from 0.05 to 0.1 mg kg-1 caused a sharp decline of Ca:Mn ratio in roots and rachises. Roots and nodes had much higher Ca:Mn ratio than rachises when soluble Cd was less than 0.5 mg kg-1 in them. These results indicate that vegetative organs have a great potential to alter more than 75% Cd into insoluble forms and increasing Ca:Mn ratio may be another way to alleviate Cd toxicity by establishing new ionic homeostasis in rice plants.

[Stabilization and long-term effect of chromium contaminated soil].

Short-term (3 d and 28 d) and long-term (1 a) stabilization effects of Cr contaminated soil were investigated through nature curing, using four amendments including ferrous sulfide, ferrous sulfate, zero-valent iron and sodium dithionite. The results indicated that ferrous sulfide and zero-valent iron were not helpful for the stabilization of Cr(VI) when directly used because of their poor solubility and immobility. Ferrous sulfate could effectively and rapidly decrease total leaching Cr and Cr(VI) content. The stabilization effect was further promoted by the generation of iron hydroxides after long-term curing. Sodium dithionite also had positive effect on soil stabilization. Appropriate addition ratio of the two chemicals could help maintain the soil pH in range of 6-8.

Effects of electrokinetic treatment of contaminated sludge on migration and transformation of Cd, Ni and Zn in various bonding states.

This study assesses the effect of electrokinetic processes on the migration and bonding states of various heavy metals in municipal sludge. The transformation and migration are discussed through the examination of sludge characteristics and distribution of Cd, Zn and Ni after electrokinetic treatments. The migration and distribution of the contaminants after the electrokinetic treatments were determined for each sludge sample by sequential extraction. The noticeable changes on the average speciation fractions of Cd, Zn and Ni were observed that oxidizable heavy metals increased and reducible fraction decreased due to the application of voltage. Bivariate correlation analysis indicated that the amounts of different bonding states of Zn and Ni were significantly correlated (P<0.05) with durations and resistance. The oxidizable Zn was negatively correlated with exchangeable and reducible Zn. Moreover, reducible Zn had a close negative relationship with residual Zn. The bonding state of Ni was significantly related to the durations of electrokinetic processes, indicating the existing of mutual transformation between different speciation fractions over time. The analysis also indicated that the exchangeable Cd showed a close negative relationship with reducible Cd (P<0.01), whereas the reducible Cd was negatively related to residual Cd (P<0.05).

[Predicting the cadmium bioavailability in the soil of sugarcane field based on the diffusive gradients in thin films with binding phase of sodium polyacrylate].

The diffusive gradients in thin films (DGT) technique with solid-state binding phases has been widely used for in situ collection and measurement of available heavy metals in waters, soils or sediments, whereas DGT with liquid binding phase is primarily used in the in situ analysis of heavy metals in waters. In this paper, rhizosphere soils of sugarcane were collected in Guangxi and the concentrations of cadmium (Cd) were determined by DGT with a solid-state binding phase of chelex100 (chelex100-DGT) and modified DGT with a liquid binding phase of sodium polyacrylate (CDM-PAAS-DGT). The result showed that the Cd contents in soils measured by DGT with both binding phases and Cd in the roots, leaves and unpolished stems of sugarcane had significant positive correlation. The extraction ability of the CDM-PAAAS-DGT was much higher than that of the chelex100-DGT. In addition, multivariate analyses were used to assess the impact of pH, cation exchange capacity (CEC), soil organic matter (OM) and texture. Two principal components were extracted and the linear regression models were established. The Cd bioavailability in soils could be accurately predicted by the CDM-PAAAS-DGT technique, which expanded its applicable area.

[Remediation of chromium (VI) contaminated soils using permeable reactive composite electrodes technology].

Electrokinetic transport processes have been shown to have potential for the effective removal of heavy metals from soils. However, pH changes near the anode and cathode limit their widespread application in the remediation of contaminated soils. Permeable reactive composite electrodes (PRCE) were made by attaching reactive materials such as Fe(0) and zeolite to the electrodes, and the effects of the composite electrodes on pH control, chromium removal efficiency and Cr speciation changes were studied in the electrokinetic remediation process of Cr( VI) contaminated soil. Composite electrodes consisting of permeable reactive materials gave better pH control and Cr removal efficiency compared to traditional electrodes, and a Fe(0) + zeolite reactive layer in the anode exhibited the best performance compared to zeolite or Fe(0) alone. After 5 days of electrokinetic remediation with a DC voltage of 2 V x cm(-1), the Fe(0) + zeolite reactive layer lowered the pH fluctuation, maintained the soil pH in the range of 5.5 to 8.5, raised the Cr(VI) removal efficiency up to 97% in any soil section, produced lower Cr(III) residues, enhanced the amount of Cr retention up to 8 and 1.8 times respectively, and transformed 98% of the Cr(VI) into lower toxicity Cr(III). This study provides a theoretical basis for the exploitation of permeable reactive composite electrodes which are a practical option for future applications.

[Stabilization of heavy metals in sewage sludge by using a ferrous iron].

The stabilization of heavy metals in sewage sludge using a ferrous iron is investigated through leaching test and fractionation analysis. The results showed that FeSO4 x 7H2O was not helpful to stabilize the heavy metals in the sludge when used individually since it reduced the pH value of sludge. The pH value dropped significantly from 6.5 to 4.4, and leaching coefficients of Zinc (Zn) and Nickel (Ni) raised, respectively, to 363.3% and 118.3% when FeSO4 x 7H2O was added individually at a ratio of 5%. However, it maintained the pH value within a slight alkaline range and the leaching coefficients of heavy metals reduced significantly when combined with Ca(OH)2 as an auxiliary agent. The pH value of sludge kept to 8.4 and leaching coefficients of Zn and Ni were reduced to 11.5% and 24.1%, for example, when 15% FeSO4 x 7H2O and 7.5% Ca(OH)2 were added together into the sludge. Besides, when Ca(OH)2 was used individually, the pH value of the sludge raised highly although the heavy metals were stabilized effectively. Under same pH value, combined addition of FeSO4 x 7H2O and Ca(OH)2 promoted the stabilization of heavy metals had a better effect than individual use of Ca(OH)2. The fractionation analysis revealed that the stable species of Zn, Cu and Pb were increased, and the instable species of Ni, Cr and Cd were also raised.

[Application of Pb stable isotope in source identification of soil pollutants].

Stable isotopes are key importance for tracing the source of heavy metals in soils. 11 surface soil samples were collected in a contaminated site according to geostatistical results, which had a series of concentrations from almost background value of Pb to more than 2 000 microg x g(-1). Pb stable isotope composition of the soil samples was analyzed to imply the sources. The results showed that 206Pb/207Pb, 208Pb/204Pb and 206Pb/204Pb of soil samples were all remarkably correlated linearly with reciprocals of Pb concentration, and 206Pb/204Pb had a significant positive linear correlation with 208Pb/204Pb, which was consistent with binary mixing model. It can be inferred that soil Pb mainly came from two sources, an anthropogenic source and soil parent material.

[Application of robust estimator in source identification of soil pollutants: a case study at a heavy metal contaminated site].

Spatial outliers can give us useful information on locations of pollution sources. A Genton's estimator based variogram was applied to analyse the concentration of Cu, Zn, Pb, Cd, As, Se and Hg in soils at a contaminated site. Spatial outliers were identified to imply the location of the pollution sources of heavy metals. The results showed that the semivariograms of 7 elements were best described by spherical model, and the nugget variances were very small, especially for Cu, Zn and Pb, which were 0.000. Compared with Matheron's estimator, Genton's estimator can damp the effect of outliers, and so get a smaller nugget effect. The spatial outliers existed in a cluster pattern, whose locations coincided with those of copper smelters in the area. This indicated that the copper smelters were the important sources of soil heavy metals.