Reactive oxygen species metabolism during the cadmium hyperaccumulation of a new hyperaccumulator Sedum alfredii (Crassulaceae)

Sedum alfredii Hance, a newly discovered hyperaccumulator, could serve as a good material for phytoremediation of Cd polluted sites. Malondialdehyde （MDA）, reactive oxygen species （ROS） and antioxidases （catalase （CAT）; superoxide dismutase （SOD）; peroxidase （POD）） in the leaf were determined when S. alfredii was treated for 15 d with various CdC12 concentrations ranging from 0 to 800 μmol/L. The results showed that the production rate of 2＇,7＇-dichlorofluorescein （DCF）, which is an indicator of ROS level, reached up to the maximum at 400 μmol/L CdCl2 and then declined with the increase of CdCl2 concentration, while MDA accumulation tended to increase. CAT activity was significantly inhibited at all tested CdCl2 concentrations and SOD activity was sharply suppressed at 800 μmol/L CdCl2. However, the enhancement of POD activity was observed when CdCl2 concentration was higher than 400 μmol/L. In addition, its activity increased when treated with 600 μmol/L CdCl2 for more than 5 d. When sodium benzoate, a free radical scavenger, was added, S. alfredii was a little more sensitive to Cd toxicity than that exposed to Cd alone, and the Cd accumulation tended to decline with the increase of sodium benzoate concentration. It came to the conclusions that POD played an important role during Cd hyperaccumulation, and the accumulation of ROS induced by Cd treatment might be involved in Cd hyperaccumulation.

Zinc adsorption and desorption characteristics in root cell wall involving zinc hyperaccumulation in Sedum alfredii Hance

Radiotracer techniques were employed to characterize 65Zn adsorption and desorption in root-cell-wall of hyperac-cumulating ecotype (HE) and non-hyperaccumulating ecotype (NHE) species of Sedum alfredii Hance. The results indicated that at the end of a 30 min short time radioisotope loading period, comparable amounts of 65Zn were accumulated in the roots of the two ecotypes Sedum alfredii, whereas 2.1-fold more 65Zn remains in NHE root after 45-min desorption. At the end of 60 min uptake period, no difference of 65Zn accumulation was observed in undesorbed root-cell-wall of Sedum alfredii. However, 3.0-fold more 65Zn accumulated in desorbed root-cell-wall of NHE. Zn2+ binding in root-cell-wall preparations of NHE was greater than that in HE under high Zn2+ concentration. All these results suggested that root-cell-wall of the two ecotypes Sedum alfredii had the same ability to adsorb Zn2+, whereas the desorption characteristics were different, and with most of 65Zn binding on root of HE being available for loading into the xylem, as a result, more 65Zn was translocated to the shoot.

Zinc removal from hyperaccumulator Sedum alfredii Hance biomass

Leaching of heavy metals from Sedum alfredii Hance biomass was studied with ammonia-ammonium chloride solution as leaching agent.The research was carried out in two phases:1)a leaching study to determine the zinc extraction efficiency of this leaching agent,and 2)a thermodynamic analysis to identify the likely reactions and stable Zn(Ⅱ)species formed in the leaching systems.Taguchi orthogonal experiment,with four variable parameters,leaching temperature,molar ratio of NH4Cl to NH3,leaching time and solid-to-liquid(L/S)ratio,and each at three levels,was used to optimize the experiment parameters by the analysis of variances.The results indicate that leaching temperature has the most dominant effect on metal extraction performance,followed by molar ratio of NH4Cl to NH3,solid-to-liquid ratio and leaching time.The optimum condition was obtained as follows:temperature of 60℃,molecular ratio of NH4Cl to NH3 of 0.6,leaching time of 2 h and solid-to-liquid ratio of 5-1.The total zinc leaching efficiency under optimum conditions reaches 97.95%.The thermodynamic study indicates that the dominant species produced by the leaching process should be the soluble Zn(NH3)4 2+.

Integrated Fertilization Regimes Boost Heavy Metals Accumulation and Biomass of Sedum alfredii Hance

The hyperaccumulator Sedum alfredii Hance(S.alfredii)may be employed for zinc(Zn)and cadmium(Cd)-polluted soil remediation.However,the low phytoremediation efficiency,related to the low biomass production,limits its use with that purpose.In this experiment,nitrogen(N),phosphorus(P),and potassium(K)fertilizers,and organic manure were applied to investigate the phytoremediation ability of S.alfredii.Hydroponic and pot experiments were conducted using Zn-Cd polluted soil.The hydroponic experiment indicated that appropriate fertilizer application could increase(p<0.05)the amount of accumulated Zn and Cd in S.alfredii.When N supply ranged from 0.5 to 2.5 mmol L^(−1),it could improve growth and accumulation of Zn and Cd in whole plants of S.alfredii.The 1 mmol L^(−1) N was an optimal N dosage for shoot biomass production and Cd accumulation in shoots,while the 2.5 mmol L^(−1) was an optimal N dosage for Zn accumulation in shoots.Both low(<0.05 mmol L^(−1))and high(>0.8 mmol L^(−1))P supply decreased growth,and Zn/Cd accumulation in whole plants of the studied species.The 0.1 mmol L^(−1) P was an optimal dosage for S.alfredii biomass production and Zn/Cd accumulation in shoots.The supply levels within the range from 0.3 to 1 mmol L^(−1) K could significantly improve the biomass production of S.alfredii and its capability to accumulate Zn and Cd in the biomass.The 0.5 mmol L^(−1) K was an optimal dosage for the whole biomass production and Zn accumulation in shoots,while the 1 mmol L^(−1) was an optimal K dosage for Zn accumulation in shoots,which was 17.2%higher than the control.Moreover,the soil pot experiment showed that the combination of organic(fermented manure)and inorganic fertilizers made significant effects on the Zn and Cd-polluted soil remediation by S.alfredii.These effects varied,however,with the application of different proportions of N,P,K and organic matter.The Zn accumulation by S.alfredii reached the highest efficiency ability under the highest fertilizer mixing rate(N:50 mg kg^(−1),P:40 mg kg^(−1),K:100 mg kg^(−1),organic matter:1%).Even more,S.alfredii showed the strongest ability to accumulate Cd with a lower fertilizer mixing rate(N:25mg kg^(−1),P:20mg kg^(−1),K:50 mg kg^(−1),organic matter:0.5%).

Organic Materials Could Improve the Phytoremediation Efficiency of Soil Potentially Hazardous Metal by Sedum alfredii Hance

Soil potentially hazardous metal(PHM)is continually attracting public attention worldwide,due to its highly toxic properties and potentially huge damage to human being through food chain.Phytoremediation is an effective and eco-friendly way in remediation technology.A pot experiment was carried out to investigate the effect of different organic materials(biogas residue(BR),mushroom residue(MR),and bamboo-shoot shell(BS))application on phytoremediation of two PHM-contaminated soils(Fuyang soil as‘heavily-polluted soil’and Wenzhou soil as‘moderately-polluted soil’,respectively)by Sedum alfrecdii Hance.The results indicated:1)for moderately-polluted soil,the 5%BR treatment had the strongest activation to Cu and Zn,for heavily-polluted soil,1%BS treatment had the highest activation effect for Cu,Zn,Pb and Cd.2)the above-ground biomass of Sedum alfredii Hance increased with the addition rate of organic materials.3)for Cd uptake of Sedum alfredii Hance in moderately-polluted soil,only 1%BS treatment had a better accumulation effect,compared to the control,for Zn element,MR treatments were weaker than the control,while other treatments were better than the control,of which 5%BR,1%BS and 5%BS accumulated more Zn element by 39.6%,32.6%and 23.8%,respectively;in heavily-polluted soil,the treatments of 5%BS,1%BR and 5%BR accumulated more Cd than the control by 12.9%,12.8%and 6.2%,respectively,the treatments with organic materials addition promoted Zn accumulation in shoots of Sedum alfredii Hance,and the best treatment was 5%BS.Therefore,an appropriate application rate of BS and BR could improve the remediation efficiency for Zn/Cd contaminated soils by Sedum alfredii Hance.

东南景天Sedum alfredii修复重金属污染土壤的研究进展

植物修复是净化重金属污染土壤最有潜力的原位修复技术之一,因地制宜地选择植物修复材料是大田植物修复成功实施的关键。国内外已发现的众多重金属超积累植物具有不同的生理特性,其中东南景天Sedum alfredii可忍耐和富集镉(Cd)和锌(Zn)等多种重金属,并且具有生长迅速的特点,是研究植物修复技术的模式植物之一。综述了东南景天对重金属的耐性、吸收、转运及解毒机制,阐明东南景天研究过程中存在的问题,并对其研究前景和方向作了简要分析,为重金属污染土壤植物修复工作的深入开展提供参考。

Accumulation of Zinc, Cadmium, and Lead in Four Populations of Sedum alfredii Growing on Lead/Zinc Mine Spoils

Sedum alfredii Hance is a newly reported zinc （Zn） and cadmium （Cd） hyperaccumulator native to China. In this study, four populations of S. alfredii were collected from Yejiwei （YJW）, Jinchuantang （JCT） and Qiaokou （QK） lead （Pb）lZn mines located in Hunan Province as well as Quzhou （QZ） PblZn mine located in Zhejiang Province for exploring the intraspecies difference of this plant in metal accumulation. Although they grew in the PblZn spoils with relatively similar levels of Zn, Cd and Pb, remarkable differences among the four populations in tissue heavy metal concentrations were observed. The shoot Zn concentration of QZ population （11 116mglkg） was highest and nearly five times higher than that of the JCT population （1 930 mg/kg）. Furthermore, the shoot Cd concentration observed in the QZ population （1 090 mg/kg） was also highest and 144 times higher than that found in the JCT population （7.5 mg/kg）. As for Pb concentrations in the shoot of different populations, a fourfold difference between the highest and the lowest was also found. Such difference on metal accumulation was population-specific and may be significantly explained by differences in the soil properties such as pH, organic matter （OM）, and electrical conductivity （EC）. Taking biomass and metal concentration in plants into consideration, the QZ, YJW and QK populations may have high potential for Zn phytoremediation, the QZ population may have the highest potential in Cd phytoremediation, and the QK population may be the most useful in Pb phytoremediation.

两种生态型东南景天(Sedum alfredii Hance)根系互作效应对植株生长及吸收利用土壤Zn的影响

采用盆栽套种方法探讨了不同程度根系互作对两种生态型东南景天的生长和吸收利用土壤Zn的影响,以及超积累生态型东南景天的根系是否能活化土壤中难利用态Zn。不同程度的根系互作采用不同隔膜处理,即无隔膜处理、400目尼龙筛布隔开和PVC板隔开。结果发现,当两种生态型东南景天套种时,不同程度的根系互作对非超积累生态型东南景天的地上部和根系的生物量、根系Zn含量有显著影响。当与超积累生态型东南景天的根系完全交互作用或采用400目尼龙筛布隔开时,非超积累生态型的根系Zn含量显著增加,其地上部和根系生物量比其单种时显著减少。这些结果间接表明,超积累生态型东南景天根系能够活化土壤中的难溶态Zn。

Effects of EDTA and plant growth-promoting rhizobacteria on plant growth and heavy metal uptake of hyperaccumulator Sedum alfredii Hance

Phytoremediation is a cost-effective and environment-friendly strategy for decontaminating heavy-metal-contaminated soil.However, the practical use of phytoremediation is constrained by the low biomass of plants and low bioavailability of heavy metals in soil.A pot experiment was conducted to investigate the effects of the metal chelator ethylenediaminetetraacetic acid(EDTA) and EDTA in combination with plant growth-promoting rhizobacteria(Burkholderia sp.D54 or Burkholderia sp.D416) on the growth and metal uptake of the hyperaccumulator Sedum alfredii Hance.According to the results, EDTA application decreased shoot and root biomass by 50% and 43%, respectively.The soil respiration and Cd,Pb, Zn uptake were depressed, while the photosynthetic rate, glutathione and phytochelatin(PC) contents were increased by EDTA application.Interestingly, Burkholderia sp.D54 and Burkholderia sp.D416 inoculation significantly relieved the inhibitory effects of EDTA on plant growth and soil respiration.Compared with the control, EDTA + D416 treatment increased the Cd concentration in shoots and decreased the Pb concentration in shoots and roots, but did not change the Zn concentration in S.alfredii plants.Furthermore,EDTA, EDTA + D54 and EDTA + D416 application increased the cysteine and PC contents in S.alfredii(p < 0.05);among all tested PCs, the most abundant species was PC2, and compared with the control, the PC2 content was increased by 371.0%, 1158.6% and 815.6%,respectively.These results will provide some insights into the practical use of EDTA and PGPR in the phytoremediation of heavy-metal-contaminated soil by S.alfredii.

Root Morphology and Zn^2＋ Uptake Kinetics of the Zn Hyperaccumulator of Sedum alfredii Hance

Root morphology and Zn^2＋ uptake kinetics of the hyperaccumulating ecotype （HE） and nonhyperaccumulating ecotype （NHE） of Sedum alfredii Hance were investigated using hydroponic methods and the radiotracer flux technique. The results indicate that root length, root surface area, and root volume of NHE decreased significantly with increasing Zn^2＋ concentration in growth media, whereas the root growth of HE was not adversely affected, and was even promoted, by 500μmol/L Zn^2＋. The concentrations of Zn^2＋ in both ecotypes of S. alfredii were positively correlated with root length, root surface area and root volumes, but no such correlation was found for root diameter. The uptake kinetics for ^65Zn^2＋ in roots of both ecotypes of S. alfredii were characterized by a rapid linear phase during the first 6 h and a slower linear phase during the subsequent period of investigation. The concentration-dependent uptake kinetics of the two ecotypes of S. alfredii could be characterized by the Michaelis-Menten equation, with the Vmax for ^65Zn^2＋ influx being threefold greater in HE compared with NHE, indicating that enhanced absorption into the root was one of the mechanisms involved in Zn hyperaccumulation. A significantly larger Vmax value suggested that there was a higher density of Zn transporters per unit membrane area in HE roots.

Chromosome doubling of Sedum alfredii Hance: A novel approach for improving phytoremediation efficiency

Sedum alfredii Hance is a cadmium(Cd)/zinc(Zn)hyperaccumulator native to China.However,its relatively low biomass restricted the large-scale application for heavy metal contamination remediation.The chromosome set doubling of S.alfredii in vitro was achieved by 0.1%–0.2%(W/V)colchicine treatment.The plant DNA ploidy was analyzed by flow cytometry and chromosome set doubling plants(CSD)were identified based on the obvious different sharp peak.A tissue culture experiment with different Cd treated levels and a field trial with natural polluted mined soil were conducted to study the effects of chromosome doubling on plant biomass and Cd accumulation in shoots.The results suggested that S.alfredii is a mixoploid.Compared with the wild type plants(WT),CSD exhibited typical"gigas"characteristics in morphology including stem thickness,root hair production,number of leaves and size of stoma guard cell.Fresh weight and dry weight of CSD were increased to 1.62–2.03-fold and 2.26–3.25-fold of WT.And Cd content of CSD showed a17.49%–42.82%increase and 59%increase under tissue culture and field condition,accordingly.In addition,the TF and in BCF of CSD were 2.37-and 1.59-fold of WT,respectively.These results proved that it is feasible to promote phytoextraction efficiency of S.alfredii in Cd contaminated soils through chromosomal engineering,which provides a novel approach for hyperaccumulator application in phytoremediation.

Heavy metal phytoextraction by Sedum alfredii is affected by continual clipping and phosphorus fertilization amendment

Improving the efficacy of phytoextraction is critical for its successful application in metal contaminated soils. Mineral nutrition affects plant growth and metal absorption and subsequently the accumulation of heavy metal through hyper-accumulator plants. This study assessed the effects of di-hydrogen phosphates （KH2PO4, Ca（H2PO4）2, NaH2PO4 and NH4H2PO4） application at three levels （22, 88 and 352 mg P/kg soil） on Sedum alfredii growth and metal uptake by three consecutive harvests on aged and Zn/Cd combined contaminated paddy soil. The addition of phosphates （P） significantly increased the amount of Zn taken up by S. alfredii due to increased shoot Zn concentration and dry matter yield （DMY） （P 〈 0.05）. The highest phytoextraction of Zn and Cd was observed in KH2PO4 and NH4H2PO4 treatment at 352 mg P/kg soil. The amount of Zn removed by phytoextraction increased in the order of 1st clipping 〈 2nd clipping 〈 3rd clipping, and for Cd extraction the order was 2nd clipping 〈 1st clipping 〈 3rd clipping. These results indicate that the application of P fertilizers coupled with multiple cuttings can enhance the removal of Zn and Cd from contaminated soils by S. alfredii, thus shortening the time needed for accomplishing remediation goals.

Combined remediation of DDT congeners and cadmium in soil by Sphingobacterium sp.D-6 and Sedum alfredii Hance

Combined pollution of 1,1,1-trichloro-2,2-bis （4-chlorophenyl） ethane （DDT） and cadmium （Cd） in agricultural soils is of great concern because they present serious risk to food security and human health.In order to develop a cost-effective and safe method for the removal of DDTs and Cd in soil,combined remediation of DDTs and Cd in soil by Sphingobacterium sp.D-6 and the hyperaccumulator,Sedum alfredii Hance was investigated.After treatment for 210 days,the degradation half-lives of DDTs in soils treated by strain D-6 decreased by 8.1% to 68.0% compared with those in the controls.The inoculation of strain D-6 into soil decreased the uptake of DDTs by pak choi and S.alfredii.The shoots/roots ratios of S.alfredii for the Cd accumulation ranged from 12.32 to 21.75.The Cd concentration in soil decreased to 65.8%-71.8% for S.alfredii treatment and 14.1%-58.2% for S.alfredii and strain D-6 combined treatment,respectively,compared with that in the control.The population size of the DDTs-degrading strain,Simpson index （1/D） and soil respiratory rate decreased in the early stage of treatment and then gradually increased,ultimately recovering to or exceeding the initial level.The results indicated that synchronous incorporation of strain D-6 and S.alfredii into soil was found to significantly （p 0.05） enhance the degradation of DDTs in soil and the hyperaccumulation of Cd in S.alfredii.It was concluded that strain D-6 and S.alfredii could be used successfully to control DDTs and Cd in contaminated soil.

Improved cadmium uptake and accumulation in the hyperaccumulator Sedum alfredii:the impact of citric acid and tartaric acid

The elucidation of a natural strategy for metal hyperaccumulation enables the rational design of technologies for the clean-up of metal-contaminated soils.Organic acid has been suggested to be involved in toxic metallic element tolerance,translocation,and accumulation in plants.The impact of exogenous organic acids on cadmium(Cd) uptake and translocation in the zinc(Zn)/Cd co-hyperaccumulator Sedum alfredii was investigated in the present study.By the addition of organic acids,short-term(2 h) root uptake of 109 Cd increased significantly,and higher 109 Cd contents in roots and shoots were noted 24 h after uptake,when compared to controls.About 85% of the 109 Cd taken up was distributed to the shoots in plants with citric acid(CA) treatments,as compared with 75% within controls.No such effect was observed for tartaric acid(TA).Reduced growth under Cd stress was significantly alleviated by low CA.Long-term application of the two organic acids both resulted in elevated Cd in plants,but the effects varied with exposure time and levels.The results imply that CA may be involved in the processes of Cd uptake,translocation and tolerance in S.alfredii,whereas the impact of TA is mainly on the root uptake of Cd.

Extraction and isolation of the salidroside-type metabolite from zinc(Zn) and cadmium(Cd) hyperaccumulator Sedum alfredii Hance

The active metabolite in the post-harvested biomass of zinc(Zn) and cadmium(Cd) hyperaccumulator Sedum alfredii Hance from phytoextraction is of great interest in China.The current study demonstrates that a salidroside-type metabolite can be yielded from the Zn/Cd hyperaccumulator S.alfredii biomass by means of sonication/ethanol extraction and macroporous resin column(AB-8 type) isolation.The concentrations of Zn and Cd in the salidroside-type metabolite were below the limitation of the national standards.

Effect of elevated CO_2 concentration on photosynthetic characteristics of hyperaccumulator Sedum alfredii under cadmium stress

The combined effects of elevated CO2 and cadmi- um （Cd） on photosynthetic rate, chlorophyll fluorescence and Cd accumulation in hyperaccumulator Sedum alfredii Hance were investigated to predict plant growth under Cd stress with rising atmospheric CO2 concentration. Both pot and hydroponic experiments were conducted and the plants were grown under ambient （350 μL L^-1） or elevated （800μL L^-1） CO2. Elevated CO2 significantly （P 〈 0.05） increased Pn （105%- 149%）, Pnmax （38.8%-63.0%） and AQY （20.0%-34.8%） of S. alfredii in all the Cd treatments, but reduced chlorophyll concentra- tion, dark respiration and photorespiration. After lo days growth in medium with 50 μM Cd under elevated CO2, PSII activities were significantly enhanced （P 〈 0.05） with Pm, Fv/ Fm, φ（Ⅱ） and qP increased by 66.1%, 7.5%, 19.5% and 16.47o, respectively, as compared with ambient-grown plants. Total Cd uptake in shoot of S. alfredii grown under elevated CO2 wasincreased by 44.1%-48-5%, which was positively correlated with the increase in Pn. These results indicate that elevated CO2 promoted the growth of S. alfredii due to increased photosynthetic carbon uptake rate and photosynthetic light- use efficiency, and showed great potential to improve the phytoextraction of Cd by 5. alfredii.

Response of ATP sulfurylase and serine acetyltransferase towards cadmium in hyperaccumulator Sedum alfredii Hance

We studied the responses of the activities of adenosine-triphosphate (ATP) sulfurylase (ATPS) and serine acetyltransferase (SAT) to cadmium (Cd) levels and treatment time in hyperaccumulating ecotype (HE) Sedum alfredii Hance, as compared with its non-hyperaccumulating ecotype (NHE). The results show that plant growth was inhibited in NHE but promoted in HE when exposed to high Cd level. Cd concentrations in leaves and shoots rapidly increased in HE rather than in NHE, and they became much higher in HE than in NHE along with increasing treatment time and Cd supply levels. ATPS activity was higher in HE than in NHE in all Cd treatments, and increased with increasing Cd supply levels in both HE and NHE when exposed to Cd treatment within 8 h. However, a marked difference of ATPS activity between HE and NHE was found with Cd treatment for 168 h, where ATPS activity increased in HE but decreased in NHE. Similarly, SAT activity was higher in HE than in NHE at all Cd treatments, but was more sensitive in NHE than in HE. Both ATPS and SAT activities in NHE leaves tended to decrease with increasing treatment time after 8 h at all Cd levels. The results reveal the different responses in sulfur assimilation enzymes and Cd accumulation between HE and NHE. With increasing Cd stress, the activities of sulfur assimilation enzymes (ATPS and SAT) were induced in HE, which may contribute to Cd accumulation in the hyperaccumulator Sedum alfredii Hance.

Differential generation of hydrogen peroxide upon exposure to zinc and cadmium in the hyperaccumulating plant specie(Sedum alfredii Hance)

Sedum alfredii Hance has been identified as zinc(Zn) and cadmium(Cd) co-hyperaccumulator.In this paper the relationships of Zn or Cd hyperaccumulation to the generation and the role of H2O2 in Sedum alfredii H.were examined.The results show that Zn and Cd contents in the shoots of Sedum alfredii H.treated with 1000 μmol/L Zn2+ and/or 200 μmol/L Cd2+ increased linearly within 15 d.Contents of total S,glutathione(GSH) and H2O2 in shoots also increased within 15 d,and then decreased.Total S and GSH contents in shoots were higher under Cd2+ treatment than under Zn2+ treatment.However,reverse trends of H2O2 content in shoots were obtained,in which much higher H2O2 content was observed in Zn2+-treated shoots than in Cd2+-treated shoots.Similarly,the microscopic imaging of H2O2 accumulation in leaves using H2O2 probe technique showed that much higher H2O2 accumulation was observed in the Zn2+-treated leaf than in the Cd2+-treated one.These results suggest that there are different responses in the generation of H2O2 upon exposure to Zn2+ and Cd2+ for the hyperaccumulator Sedum alfredii H.And this is the first report that the generation of H2O2 may play an important role in Zn hyperaccumulation in the leaves.Our results also imply that GSH may play an important role in the detoxification of dissociated Zn/Cd and the generation of H2O2.

Compounded chelating agent derived from fruit residue extracts effectively enhances Cd phytoextraction by Sedum alfredii

Chelating agent is known as the enhancer for metal phytoextraction;however,there is still a lack of efficient and environmentally sustainable chelators.Here,lemon residue extraction(LRE),prepared from 11 kinds of fruit wastes,was combined with N,N-bis(carboxymethyl)glutamic acid(GLDA),and tea saponin(T.S.)for the compounded plant-derived chelator(CPC),and their influences on Cd phytoextraction by the hyperaccumulator Sedum alfredii was evaluated.Among these fruits,the lemon residue extracted the most significant amount of Cd from the soil.The most effective CPC was at the volume ratio of three agents being 15:4:1(LRE:GLDA:T.S.).Compared with the deionized water,the solubility of three Cd minerals was increased by 85–256 times,and Cd speciation was substantially altered after CPC application.In the pot experiment,CPC addition caused evident increases in plant shoot biomass,Cd phytoextraction efficiency,and organic matter content compared with EDTA and nitrilotriacetic acid(NTA)application.CPC induced fewer changes in bacterial community composition compared with EDTA and had no pronounced influence on microbial biomass carbon and bacterialα-diversity,suggesting CPC had a subtle impact on the microbiological environments.Our study provides a theoretical base for the reutilization of fruit wastes and the development of environmental-friendly chelator that assists Cd phytoextraction.