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)/Zn mines located in Hunan Province as well as Quzhou (QZ) Pb/Zn mine located in Zhejiang Province for exploring the intraspecies difference of this plant in metal accumulation. Although they grew in the Pb/Zn 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 (11 116 mg/kg) was highest and nearly five times higher than that of the JCT population (1930 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 opulation-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.     

Effect of simultaneous establishment of Sedum alfredii and Zea mays on heavy metal accumulation in plants

Land application of biosolids to improve agricultural productivity is a cost-effective approach for resource recovery. Unfortunately, municipal biosolids often contain high concentrations of heavy metals, including zinc and copper. In this study, a co-cropping technique was investigated using a known zinc hyperaccumulator, Sedum alfredii with a grain crop, Zea mays. After a 3-mo growth trial, the results indicate that when Z. mays is co-cropped with S. alfredii, heavy metals accumulated in the grains were significantly reduced when compared to monoculture cropping. Co-cropping improved the growth of both plant species. In addition, the biosolids maintained stable pH, N-P-K concentrations, germination potential, and water content after the plant treatment, regardless of the plant species used in the trial. In conclusion, co-cropping with hyperaccumulators may be an effective approach to reducing the risk of contaminant uptake in edible crops.     

Cellular sequestration of cadmium in the hyperaccumulator plant species Sedum alfredii

Spatial imaging of cadmium (Cd) in the hyperaccumulator Sedum alfredii was investigated in vivo by laser ablation inductively coupled plasma mass spectrometry and x-ray microfluorescence imaging. Preferential Cd accumulation in the pith and cortex was observed in stems of the Cd hyperaccumulating ecotype (HE), whereas Cd was restricted to the vascular bundles in its contrasting nonhyperaccumulating ecotype. Cd concentrations of up to 15,000 μg g(-1) were measured in the pith cells, which was many fold higher than the concentrations in the stem epidermis and vascular bundles in the HE plants. In the leaves of the HE, Cd was mainly localized to the mesophyll and vascular cells rather than the epidermis. The distribution pattern of Cd in both stems and leaves of the HE was very similar to calcium but not zinc, irrespective of Cd exposure levels. Extended x-ray absorption fine structure spectroscopy analysis showed that Cd in the stems and leaves of the HE was mainly associated with oxygen ligands, and a larger proportion (about 70% in leaves and 47% in stems) of Cd was bound with malic acid, which was the major organic acid in the shoots of the plants. These results indicate that a majority of Cd in HE accumulates in the parenchyma cells, especially in stems, and is likely associated with calcium pathways and bound with organic acid (malate), which is indicative of a critical role of vacuolar sequestration of Cd in the HE S. alfredii.     

Abscisic acid‐mediated modifications of radial apoplastic transport pathway play a key role in cadmium uptake in hyperaccumulator Sedum alfredii

Abscisic acid (ABA) is a key phytohormone underlying plant resistance to toxic metals. However, regulatory effects of ABA on apoplastic transport in roots and consequences for uptake of metal ions are poorly understood. Here, we demonstrate how ABA regulates development of apoplastic barriers in roots of two ecotypes of Sedum alfredii and assess effects on cadmium (Cd) uptake. Under Cd treatment, increased endogenous ABA level was detected in roots of nonhyperaccumulating ecotype (NHE) due to up‐regulated expressions of ABA biosynthesis genes (SaABA2, SaNCED), but no change was observed in hyperaccumulating ecotype (HE). Simultaneously, endodermal Casparian strips (CSs) and suberin lamellae (SL) were deposited closer to root tips of NHE compared with HE. Interestingly, the vessel‐to‐CSs overlap was identified as an ABA‐driven anatomical trait. Results of correlation analyses and exogenous applications of ABA/Abamine indicate that ABA regulates development of both types of apoplastic barriers through promoting activities of phenylalanine ammonialyase, peroxidase, and expressions of suberin‐related genes (SaCYP86A1, SaGPAT5, and SaKCS20). Using scanning ion‐selected electrode technique and PTS tracer confirmed that ABA‐promoted deposition of CSs and SL significantly reduced Cd entrance into root stele. Therefore, maintenance of low ABA levels in HE minimized deposition of apoplastic barriers and allowed maximization of Cd uptake via apoplastic pathway.     

超积累植物伴矿景天镉耐受基因SpMT2的分离及功能鉴定

超积累植物由于其对重金属具有地上部超积累以及超耐受等特性,不仅是研究植物离子转运及毒性耐受的理想模式,而且在植物修复的发展和应用中具有不可替代的作用。伴矿景天是近年在我国境内发现的一种景天科镉(Cd)/锌(Zn)超积累植物。为鉴定其富集和耐受Cd的关键基因,笔者构建了其酵母表达cDNA文库,利用酵母的遗传互补系统筛选到一个极大提高了酵母抗Cd能力的基因SpMT2。SpMT2属于富含半胱氨酸(Cys)的金属硫蛋白(Metallothionein)家族。亚细胞定位表明SpMT2表达于酵母细胞质中,并特异地提高酵母对Cd的抗性。进一步研究发现SpMT2的表达显著降低了酵母液泡中Cd含量,但酵母吸收的总Cd含量无显著变化。推测抗性增加是由于SpMT2在酵母细胞质中通过螯合Cd从而降低Cd对酵母的毒害。qRT-PCR分析表明SpMT2在伴矿景天的根和地上部都高丰度表达,且不受Cd诱导变化。鉴于SpMT2也定位于植物细胞质中,结合上述结果,推测SpMT2可能在伴矿景天细胞质中螯合Cd,在降低Cd毒害的同时可能还保持Cd在细胞质中的流动性,从而在Cd长途转运过程中也发挥重要作用。     

Detection of phytochelatins in the hyperaccumulator Sedum alfredii exposed to cadmium and lead

Phytochelatins (PCs) are known to play an essential role in the heavy metal detoxification of some higher plants and fungi by chelating heavy metals. However, three recent papers reported that no PCs could be detected in the hyperaccumulator Sedum alfredii Hance upon cadmium, lead or zinc treatment, respectively. In this paper, PC synthesis was assayed again in the mine population of S. alfredii with the help of reversed phase high-performance liquid chromatography (HPLC), HPLC-mass spectrometry, and HPLC-tandem mass spectrometry. Our data showed that PC formation could be induced in the leaf, stem and root tissues of S. alfredii upon exposure to 400 microM cadmium, and only in the stem and root when exposed to 700 microM lead. However, no PCs were found in any part of S. alfredii when it was subjected to exposure to 1600 microM zinc.     

Cleaning up of heavy metals-polluted water by a terrestrial hyperaccumulator Sedum alfredii Hance

Sedum alfredii Hance is a terrestrial zinc/cadmium （Zn/Cd）-hyperaccumulating and lead （Pb）-accumulating plant. Previous studies on S. alfredii were mostly focused on its physiological mechanism of heavy metal uptake and the application in phytoextraction of metals from contaminated soils. In this study, we evaluated the application potential of S. alfredii in the cleanup of heavy metals from contaminated lake water. Our research revealed that changing pH in lake water would not make particular difference on the final accumulation amount of heavy metals, because the acidic water environment negatively affected plant growth compared with the neutral and alkaline environments, but was more conducive for heavy metal absorption and accumulation. In addition, S. alfredii showed an increase of approximately 2.2-fold in dry weight （DW） when cultured with lake water for 25 d. At the same time, it accumulated approximately 5.0 mg/kg DW of Cd and 41.4 mg/kg DW of Pb. The absorption of heavy metals was highly effective during the first 10 d of culture. Also, the quality of lake water was greatly improved after only 2-d cleanup by S. aifredii. In general, this hyperaccumulator exhibits great potential for application in the cleanup of heavy metals-polluted waters.     

The effects of heavy metal pollution on genetic diversity in zinc/cadmium hyperaccumulator Sedum alfredii populations

The genetic diversity and population structure of seven populations of Sedum alfredii growing in lead/zinc (Pb/Zn) mine spoils or in uncontaminated soils from eastern and southern China were investigated using random amplified polymorphic DNA (RAPD) technology. Four of the sampled sites were heavily contaminated with heavy metals (Zn, Cd, Pb), and extremely high concentrations of Zn, Cd, and Pb were found among these corresponding populations. A significant reduction of genetic diversity was detected in the mining populations. The reduction of genetic diversity could be derived from a bottleneck effect and might also be attributed to the prevalence of vegetative reproduction of the mining populations. Analysis of molecular variance (AMOVA) and the unweighted pair group method with arithmetic mean (UPGMA) tree derived from genetic distances further corroborated that the genetic differentiation between mine populations and uncontaminated populations was significant. Polymorphism with the heavy metal accumulation capability of S. alfredii probably due to the genetic variation among populations and heavy metal contamination could have more impact on the genetic diversity and population structure of S. alfredii populations than geographic distance.     

Nickel levels in arthropods associated with Ni hyperaccumulator plants from an ultramafic site in New Caledonia

Arthropods (mainly insects) were collected from a forest site that contained at least six species of Ni hyperaccumulators. Whole body Ni analysis was performed for 12 arthropod taxa, two of which were studied at different life cycle stages. We found two Nitolerant insects. The pentatomid heteropteran Utana viridipuncta, feeding on fruits of the Ni hyperaccumulator Hybanthus austrocaledonicus, contained a mean of 2 600 μNi/g in nymphs and 750 μNi/g in adults. The tephritid fly Bactrocera psidii, feeding on pulp of Sebertia acuminata fruits that contained 6 900 μNi/g, contained 420 μNi/g as larvae that had evacuated their guts and significantly less (65 μNi/g) as adults. European honeybees (Apis mellifera) visiting flowers of the Ni hyperaccumulator H. austrocaledonicus contained significantly more Ni (8‐fold more) than those collected from flowers of Myodocarpus fraxinifolius, a non‐hyperaccumulator. Our results show that some insects feed on Ni hyperaccumulator plants and that their feeding mobilizes Ni into local food webs.     

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

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

古老铅锌矿区生态型东南景天对锌耐性及超积累特性的研究

 植物长期生长在重金属污染的生境中,逐渐进化成不同的生态型。通过调查中国东南部古老Pb/zn矿和非矿山生境中的植物种群,发现生长在古老Pb／Zn矿的东南景天(Sedum alfredii Hance)是一种新的Zn超积累植物。在自然和控制条件下,古老Pb／Zn矿生态型比非矿山生态型植株的茎粗、叶片大、植株高。在矿山土壤Zn有效含量为105.5～325.4mg·kg-1时,矿山生态型东南景天植株地上部Zn含量为4134～5000mg·kg-1;当营养液中Zn浓度为1223.6μmol时,其Zn含量高达2％。在相同Zn浓度下,矿山生态型地上部Zn含量比非矿山生态型高30倍左右。两种生态型体内Zn分布也不同,古老铅锌矿山生态型的不同器官中Zn含量以茎>叶片>根系,而非矿山生态型则以根系>茎>叶片。古老铅锌矿山生态型地上部积累的Zn占植株总积累量的90％以上,其中叶片和茎分别占41.66％±5.46％和54.75％±5.87％;非矿山生态型各器官中积累的Zn远远低于古老铅锌矿山生态型,各器官中积累的Zn以茎>根系>叶片。本研究表明,这两种生态型东南景天的发现,为今后探讨植物耐高Zn胁迫和超积累Zn的微进化过程提供了非常有价值的材料,也为Zn污染土壤的植物修复提供了一种很有潜力的候选材料。     

Cadmium hyperaccumulation leads to an increase of glutathione rather than phytochelatins in the cadmium hyperaccumulator Sedum alfredii

Sedum alfredii has been reported to be a cadmium (Cd) hyperaccumulator. Phytochelatins (PCs) and other thiol (SH)-containing compounds have been proposed to play an important role in the detoxification and tolerance of some heavy metals, but it is not clear whether PCs are responsible for Cd hyperaccumulation and tolerance in S. alfredii. In this study, two geographically isolated populations of S. alfredii were studied: one population grew on an old Pb/Zn mine site, while the other on a non-mine site. The mine population of this species exhibited a stronger heavy metal tolerance than in the other population. Root-to-shoot transport of Cd was higher in population located at the mine site than at the non-mine site. Considerable amounts of Cd were accumulated in leaves and stems of mine plants, while most Cd was distributed in roots of non-mine plants. Non-protein SH in plant tissues of two populations were further investigated by a HPLC pre-column derivatization system. Upon exposure to Cd, no PCs were detected in all tissues of mine population, while an appreciable amount of glutathione (GSH) was observed in the descending order of stem>root>leaf. The concentrations of GSH consistently increased with the increase of exogenous Cd concentrations and time. On the contrary, Cd exposure strongly induced the production of PCs (mainly PC(2) and PC(3)) and GSH in plant tissues of non-mine population, and the concentrations of GSH showed an initial drop over the duration of 7-d exposure. The present results provided strong evidence that PCs are not involved in Cd transport, hyperaccumulation and tolerance in mine population of S. alfredii.     

镉锌超积累植物伴矿景天焚烧飞灰的重金属浸出特性及安全处置

为探究超积累植物伴矿景天焚烧灰浸出回收有价金属的可行性,以800℃流化床焚烧旋风灰为材料,研究盐酸、硝酸、氯化铵以及不同浓度、液固比和浸出时间对飞灰中重金属浸出规律的影响,同时评价浸出残渣的安全性.结果 表明:浸提剂种类、浓度及液固比是影响浸出的重要参数,浸出时间的影响较小,酸性浸提剂浸出能力明显强于氯化铵盐溶液浸出;...     

Cadmium resistant Enterobacter cloacae and Klebsiella sp. isolated from industrial effluents and their possible role in cadmium detoxification

Three bacterial strains, two of Klebsiella sp. and one Enterobacter cloacae were isolated from industrial effluents of chemical and textile industries. They showed high efficiency in removing cadmium (Cd²⁺) from the medium. When 100 g/ml of Cd was added to the medium, the three isolates namely CMBL-Cd1, CMBL-Cd2 and CMBL-Cd3 removed or accumulated 86%, 87% and 85% of Cd, respectively, from the medium within 24h. Plasmids were detected in all the three strains. Plasmids of E. cloacae (pCBL1) and Klebsiella sp. (pCBL2 and pCBL3), estimated to be 6.6kb, were used to transform Escherichia coli C600. The transformed E. coli cells showed elevated resistance to Cd. Ethidium bromide curing indicated the presence of the Cd resistance gene on the plasmid. Resistance of the isolated strains against other metals like chromium (cr⁶⁺) and lead (pb²⁺) and a number of antibiotics was also checked. Cured strains showed lowered resistance against Cr and some antibiotics. This again supported the indication of the presence of Cd, Cr and some antibiotics resistance genes on plasmids.     

Increase of glutathione in mine population of Sedum alfredii: a Zn hyperaccumulator and Pb accumulator

Phytochelatins (PCs) have been induced in a large range of plant species, but their role in heavy metal tolerance is unclear. Sedum alfredii is a new zinc (Zn) hyperaccumulator and lead (Pb) accumulator found in an old Pb/Zn mine in the Zhejiang Province of China. Until now, the mechanisms of its hyperaccumulation/accumulation and tolerance were poorly understood. The aim of this work was to investigate whether PCs were differentially produced in mine populations of S. alfredii compared with a non-mine control of the same species. The results showed that plants from the mine site were more tolerant to increasing Zn and Pb concentrations than those from the control site. No PCs and cysteine (Cys) were detected by pre-column derivatization with HPLC fluorescence in any tissues of two populations at any treatment, which in turn indicated they were not responsible for Zn and Pb tolerance in the mine population. Instead, Zn and Pb treatments resulted in the increase of glutathione (GSH) for both populations in a tissue-dependent manner. Significant increases were observed in leaf, stem and root tissues of plants grown on the mine site. The results suggest that GSH, rather man PCs, may be involved in Zn and Pb transport, hyperaccumulation/accumulation and tolerance in mine population of S. alfredii.     

植物重金属超富集机理研究进展

植物超富集重金属机理主要涉及植物对金属离子高的吸收、运输能力,区域化作用及螯合作用等方面,其中跨膜运载蛋白的表达、调控对重金属超富集这一特性起了关键作用。金属阳离子运载蛋白家族主要包括CDF家族、NRAMP家族和ZIP家族等,在超富集植物中已克隆出多个家族的金属运载蛋白基因,这些基因的过量表达对重金属在细胞中的运输、分布和富集及提高植物的抗性方面发挥了重要作用。综述了近年来研究重金属超富集植物吸收、转运和贮存Zn、Ni、Cd等重金属的生理和分子机制所取得的主要进展。     

Characterization of a High Affinity Phytochelatin Synthase from The Cd‐Utilizing Marine Diatom Thalassiosira pseudonana

Phytochelatin synthase (PC synthase) is the enzyme that catalyzes the production of phytochelatins, peptides of the structure (γ‐Glu‐Cys)_n‐Gly, where n = 2–11, from the sulfhydryl‐containing tripeptide glutathione, in response to elevated metal exposure. Biochemical utilization of Cd in the marine diatom Thalassiosira weissfloggi, as well as unusually high ratios of PC to Cd in some Thalassiosira species including T. pseudonana Hasle et Heimdal, motivated the characterization of T. pseudonana PC synthase 1 (TpPCS1). This enzyme is the product of one of three genes in the T. pseudonana genome predicted to encode for a PC synthase based on its homology to canonical PC synthases previously examined. TpPCS1 was cloned, expressed in Escherichia coli and purified under both aerobic and anaerobic conditions. TpPCS1 exhibits several characteristics that set it distinctly apart from the well‐studied PC synthase, Arabidopsis thaliana PCS1 (AtPCS1). It is extremely sensitive to oxidation, which suppresses activity, and it is readily inhibited by the addition of Cd in the absence of thiolate ligands. TpPCS1 also has significantly greater affinity for one of its key substrates, the bis‐glutathionato‐Cd complex. TpPCS1 kinetics is best described by a ternary complex model, as opposed to the ping‐pong model used to describe AtPCS1 kinetics. The findings indicate that although the function of TpPCS1 is synonymous to that of AtPCS1, its divergent biochemistry suggests adaptation of this enzyme to the distinct trace metal chemistry of the marine environment and the unique physiological needs of T. pseudonana.