Growth Response and Metal Accumulation of Sedum alfredii to Cd/Zn Complex-Polluted Ion Levels

Sedum alfredii Hance has been identified as a new Zn-hyperaccumulator native to China. In this study, responses and metal accumulation of S alfredii were examined under Zn/Cd complex polluted conditions. The results showed that optimal growth of S alfredii in terms of the maximum dry matter yield was observed at Zn/Cd complex level of 500/100 mumol/L. Plant cadmium (Cd) or zinc (Zn) concentrations increased with increasing Cd or Zn supply. During the 20 d treatment, the highest Cd concentration in the leaves reached 12.1 g/kg at Zn/Cd level of 50/400 mumol/L and that of Zn in the stems was 23.2 g/kg at Zn/Cd level of 1000/50 mumol/L. The distribution of Cd in different plant parts decreased in the order: leaf > stem greater than or equal to root, whereas that of Zn was: stem > leaf greater than or equal to root. The accumulation of Cd and Zn in the shoots and roots of S. alfredii increased with the increasing of Zn/Cd supply levels, peaked at Zn/Cd levels of 250/400 and 500/100 mumol/L, respectively. The highest Cd and Zn uptake by the shoots was approximately 5 and 11 mg/plant, and was over 20 and 10 times higher than those in the roots, respectively. Zn supply at levels less than or equal to 500 mumol/L increased plant Cd concentrations, whereas high Zn supply decreased root Cd but did not affect leaf Cd concentrations in S alfredii Low Cd supply increased Zn concentration in the leaves, but Cd supply higher than 50 mumol/L considerably reduced root Zn concentrations, especially at low Zn level. These results indicate that S. alfredii can tolerate high Zn/Cd complex levels and has an extraordinary ability to hyperaccumulate not only Zn but also Cd. It could provide a new valuable plant material for understanding the mechanisms responsible for co-hyperaccumulation of Zn and Cd as well as for phytoremediation of the Cd/Zn complex polluted soils.

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。

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.

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%).

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.

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+.

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.

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.

Sedum alfredii : A New Lead-Accumulating Ecotype

hi a survey of plant population, Sedum alfredii Hance, a new lead (Pb)-tolerant and lead (Pb)accumulating ecotype, was found in an old Pb/Zn mining area in Zhejiang Province of China. The growth and Pb content of plant ecotypes being able to and unable to accumulate Pb were studied by hydroponic culture with different concentrations of Pb(NO3)(2). Growth of shoots of accumulating ecotype was not affected by Ph treatments up to 320 mg/L, whereas that of non-accumulating ecotype was inhibited in all Ph treatments. The Ph concentrations in the roots and shoots of accumulating ecotype increased with increasing of Pb level in the nutrient solution. The maximum Ph concentrations in the shoots and roots of accumulating ecotype were 514 mg/kg and 13 922 mg/kg, 2.27 times and 2.62 times as much as that of non-accumulating ecotype, respectively. The highest rate of Pb accumulation of accumulating ecotype was 8.62 mug/plant/d, 7.16 times as much as that of non-accumulating ecotype. Due to its fast growth rate and high Pb-accumulating ability, from a phytoremediation perspective, accumulating ecotype of S. alfredii is a potential plant species for Pb removal from contaminated soils.

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。

Subcellular Distribution of Cadmium in Mining Ecotype Sedum alfredii

The mining ecotype Sedum alfredii Hance could tolerate and grow normally in a nutritive solution containing cadmium (Cd) as high as 400 mumol/L. Under such a high Cd concentration, the subcellular accumulation of Cd in root, stem and leaf of this plant was found to be the highest in the cell wall, less in the soluble fraction and lowest in the cell organs. The mode of subcellular distribution of Cd in the mining ecotype S. alfredii was similar to other hyper accumulators of heavy metals, in which Cd was distributed more in the aerial part of plant. The results suggest that the mining ecotype S. alfredii is a new species of Cd hyperaccumulator.

Zn Accumulation and Subcellular Distribution in the Zn Hyperaccumulator Sedurn alfredii Hance

Zn accumulation and subcellular distribution in leaves of the hyperaccumulating ecotype （HE） and non-hyperaccumulating ecotype （NHE） of Sedum alfredii Hance were studied using radiotracer and gradient centrifugation techniques. Leaf Zn accumulation in the HE of S. alfredii was 18.5-26.7 times greater than that in the NHE when the plants were grown at 1-500μmol Zn L-1. Leaf section uptake of 65Zn was highly dependent on external Zn levels. Greater 65Zn uptake in HE was noted only at external Zn levels 〉 100μmol L-1. Zinc subcellular distribution in the leaves of the two ecotypes of S. alfredii was： cell wall 〉 soluble fraction 〉 cell organelle. However, more Zn was distributed to the leaf cell wall and soluble fractions for HE than for NHE. In the leaf of HE, 91%-94% of the Zn was found in the cell walls and the soluble fraction and only 6%-9% Zn was distributed in the cell organelle fraction. For NHE, about 20%-26% Zn was recovered in the cell organelle fraction. In stems, Zn distribution to the ceil wail fraction was approximately two fold greater in the HE than that in the NHE. For the hyperaccumulating ecotype of S. alfredii, the cell wall and the vacuole played a very important role in Zn tolerance and hyperaccumulation.

Feasibility of enhanced phytoextraction of Zn contaminated soil with Zn mobilizing and plant growth promoting endophytic bacteria

Three bacterial endophytes of Sedum alfredii, VI8L2, II8L4 and VI8R2, were examined for promoting soil Zn bioavailability and Zn accumulation in S. alfredii. Results showed that three strains were re-introduced into S. alfredii rhizosphere soils under Zn stress and resulted in better plant growth, as roots biomass increased from 80% to 525% and shoot biomass from 11% to 47% compared with the uninoculated ones. Strains IVsLz, II8L4 and IVsR2 significantly increased shoot and root Zn concentrations in the ZnCO3 contaminated soil. Inoculation with strain IVsL2 resulted in 44% and 39% higher shoot and root Zn concentrations, while strain IV8R2 significantly decreased shoot Zn concentration in the Zn3（PO4）2 contaminated soils. In the aged contaminated soil, isolates IVsL2, IIsL4 and IVsR2 significantly increased root Zn concentration, but decreased shoot Zn concentration of Sedum alfredii. It suggested that endophytes might be used for enhancing phytoextraction efficiency.

Differences of Uptake and Accumulation of Zinc in Four Species of Sedum

Four species of Sedum L. were grown in nutrient solution for the comparison of their Zn uptake and accumulation. S. alfredii Hance showed much greater tolerance to Zn than the other three species. Shoot and root yields of S. sarmentosum Bunge, S. bulbiferum Makino, and S. emarginatum Migo decreased with increasing Zn concentration in the solution, while shoot and root yields of S. alfredii increased when Zn concentration was ≤80 mg·L -1. At 80 mg·L -1 Zn, Zn concentration in shoots of S. alfredii reached 19.09 mg·g -1. S. alfredii was also more efficient in Zn translocation from roots to shoots, while Zn concentration in shoots was much higher than in roots. However, this was not the case for the other three species. The results showed that S. alfredii is a Zn hyperaccumulator and could be useful for the phytoremediation of Zn-contaminated soils.

不同pH值混合螯合剂对土壤重金属淋洗及植物提取的影响

为了得出混合螯合剂(MC)淋洗去除重金属的最佳pH值以及对后续植物提取重金属的影响,用Ca(OH)2将pH值为2.75的MC提高至pH值5、7和9,对重金属污染土壤进行了盆栽淋洗试验,而后种植东南景天(Sedum Alfredii),测定淋出液及植物重金属含量。另外,通过浸提试验研究了含有不同阳离子的MC对重金属淋洗效果的影响。结果表明,pH值为5和7的MC显著提高了Cd、Pb和Cu的淋出率。与Na+、K+离子相比,Ca2+的存在能够提高MC对重金属的淋洗去除。pH值7和9的MC淋洗土壤后提高了东南景天的生物量,但是降低东南景天中Cd和Zn的浓度,导致其植物提取率低于无淋洗剂对照。在化学淋洗+植物提取联合技术中,Cd和Zn主要靠植物提取去除,植物提取率分别可达土壤Cd的30%～40%和土壤Zn的6.5%～6.9%;而Pb和Cu主要靠混合螯合剂淋洗去除,去除率分别为2.3%～2.6%和1.6%～2.0%。综合来说,如果需要同时去除Cd、Zn、Pb和Cu,降低土壤重金属有效态含量,用pH值9的MC淋洗土壤联合植物提取较为合适。

东南景天捕光叶绿素a/b结合蛋白基因SaLhcb2的分离及功能

LHCB基因对植物适应各种环境胁迫的过程中起着重要作用。序列分析显示:东南景天Sedum alfredii的Sa Lhcb2基因全长929 bp,其中开放阅读框(ORF)为798 bp,含有1个74 bp的内含子。通过蛋白序列比对,Sa Lhcb2基因编码的蛋白与多种植物的蛋白序列同源性都很高(92%以上)。分析400μmol·L-1镉离子(Cd2+),铜离子(Cu2+)和铅离子(Pb2+)胁迫处理后的东南景天,结果显示:镉处理后Sa Lhcb2基因在茎、叶中表达量快速上升(12.0h内),根中表达量到48.0 h后才上调。铜处理后0.5 h根中表达显著上调,胁迫6.0 h后茎中表达量显著上调,随后一直降低,叶片中该基因表达量一直较低。铅处理后,根中表达量降低,96.0 h左右比对照略微上调,而茎中96.0 h内表达量相比对照都上调,叶片中96.0 h内都降低。研究结果表明:Sa Lhcb2与东南景天的镉、铜、铅胁迫抗性有密切的相关性。

土壤pH对东南景天修复镉污染土壤的影响研究

东南景天是一种镉和锌的超积累植物,改变土壤p H能否有效提高其吸收镉的效率,需要进一步验证。采用盆栽实验研究不同土壤p H下东南景天吸收和积累Cd的差异以及对Ca Cl2提取有效态镉的影响。结果表明,降低土壤p H值显著提高了土壤镉的有效态含量。弱酸性土壤即p H接近5.5时东南景天生物量及累积镉的量最大,土壤镉去除率也最高,达6.6%。强酸性即当p H接近4时,虽然植物地上与地下镉含量均最高,但生物量最小,植物去除率较其他处理低。研究证实降低土壤p H是提高植物提取效率的有效办法,这为进一步利用东南景天修复镉污染土壤,提高修复效率提供了科学依据。

Selection of appropriate organic additives for enhancing Zn and Cd phytoextraction by hyperaccumulators

Chelant-enhanced phytoextraction is one of the most promising technologies to remove heavy metals from soil. The key of the technology is to choose suitable additives in combination with a suitable plant. In the present study, laboratory batch experiment of metal solubilization, cress seeds germination were undertaken to investigate the metal-mobilizing capability and the phytotoxicity of organic additives, including ethylene diamine triacetic acid （EDTA）, citric acid, acetic acid, oxalic acid, glutamine and monosodium glutamate waste liquid （MGWL） from food industry. Experiments in pots were carried out to study the effects of the additives on Zn and Cd phytoextraction. Furthermore, a leaching experiment with lysimeter was performed to evaluate the environmental risks of additive-induced leaching to underground water. The results showed that EDTA had a strong mobilizing ability for Zn and Cd, followed by mixed reagent （MR） and MGWL. MGWL and acetic acid at 5 mmol equivalent per liter resulted in seed germination index less than 2%. Experiments in pots verified the phytotoxicity of acetic acid and MGWL. Addition of the mixed reagent at 6--10 mmol/kg significantly increased Zn phytoextraction by Thlaspi caerulescens. The same for EDTA and the mixed reagent at 10 mmol/kg by Sedum dfredii. But only mixed reagents could significantly increase Cd phytoextraction by the studied hyperaccumulators. This suggested that the strong chelant was not always the good agent to enhance phytoextraction. S. alfredii combined with 2--10 mmol/kg soil MR was preferred for phytoremediation of Cd/Zn contaminated soils in southern China, this could result in high phytoextraction of Cd/Zn and reduce the leaching risk to underground water than EDTA assisted phytoextration.

植物套作系统修复镉污染农田土壤的效应——以苏南地区为例

为了研究不同套作模式对苏南地区Cd污染农田土壤的修复效果,选用能在苏南地区生长的伴矿景天、东南景天、苋菜和龙葵4种Cd超富集植物与大豆、油菜和小麦3种常规作物在Cd污染农田土壤中进行单作和套作修复试验.结果显示,与油菜套作后,4种超富集植物的Cd吸收量均有所增加,其中东南景天增幅最大,地上部Cd含量比单作增加24.4%;与小麦套作后,东南景天对Cd的吸收有所增加,地上部Cd含量比单作增加2.75%,龙葵和苋菜对Cd的吸收变化不显著,说明与油菜和小麦套作均能促进东南景天对Cd的吸收.套作伴矿景天减少了大豆根部Cd含量,套作苋菜和龙葵显著降低大豆茎、叶部分的Cd含量;套作模式下油菜根、茎的Cd含量均显著降低,叶片Cd含量受套作影响不显著;套作均显著降低了小麦的Cd含量,表明套作能抑制作物对Cd的吸收.此外,东南景天-油菜模式下Cd总积累量和土壤Cd去除率最高,为1343μg/盆和16.4%;东南景天-小麦次之,为1053μg/盆和9.8%.因此,东南景天-油菜/小麦两种套作模式均是苏南地区Cd污染农田土壤的适宜植物套作修复模式.研究结果可为该地区Cd污染农田修复同时安全利用提供科学依据和技术支持.