Cadmium and copper uptake and accumulation by Sesbania rostrata seedling, a N-fixing annual plant: implications for the mechanism of heavy metal tolerance

Sesbania rostrata,an annual tropical legume,has been found to be tolerant to heavy metals,with an unknown mechanism.It is a promising candidate species for revegetation at mine tailings.In this study,sequential extractions with five buffers and strong acids were used to extract various chemical forms of cadmium and copper in S.rostrata,with or without Cd or Cu treatments,so that the mechanisms of tolerance and detoxification could be inferred.Both metals had low transition rates from roots to the aboveground of S.rostrata.The transition ratio of Cd (4.00%) was higher than that of Cu (1.46%).The proportion of NaCl extracted Cd (mostly in proteinbinding forms) increased drastically in Cd treated plants from being undetectable in untreated plants.This suggests that Cd induced biochemical processes producing proteinlike phytochelatins that served as a major mechanism for the high Cd tolerance of S.rostrata.The case for Cu was quite different,indicating that the mechanism for metal tolerance in S.rostrata is metal-specific.The proportion of water-insoluble Cu (e.g.oxalate and phosphate) in roots increased significantly with Cu treatment,which partially explains the tolerance of S.rostrata to Cu.However,how S.rostrata copes with the high biotic activity of inorganic salts of Cu,which increased in all parts of the plant under Cu stress,is a question for future studies.Sesbania rostrata is among the very few N-fixing plants tolerant to heavy metals.This study provides evidence for the detoxification mechanism of metals in Sesbania rostrata.     

Phytochelatin is not a primary factor in determining copper tolerance

Phytochelatin (PC) is involved in the detoxification of harmful, non-essential heavy metals and the homeostasis of essential heavy metals in plants. Its synthesis can be induced by either cadmium (Cd) or copper (Cu), and can form stable complexes with either element. This might suggest that PC has an important role in determining plant tolerance to both. However, this is not clearly apparent, as evidenced by a PC-deficient and Cd-sensitiveArabidopsis mutant (cad1-3) that shows no significant increase in its sensitivity to copper. Therefore, we investigated whether the mechanism for Cu tolerance differed from that for Cd by analyzing copper sensitivity in Cd-tolerant transgenics and Cd-sensitive mutants ofArabidopsis. Cadmium-tolerant transgenic plants that over-expressedA. thaliana phytochelatin synthase 1 (AtPCS1) were not tolerant of copper stress, thereby supporting the hypothesis that PC is not primarily involved in this tolerance mechanism. We also investigated Cu tolerance incad2-1, a Cd-sensitive and glutathione (GSH)-deficientArabidopsis mutant. Paradoxically,cad2-1 was more resistant to copper stress than were wild-type plants. This was likely due to the high level of cysteine present in that mutant. However, when the growth medium was supplemented with cysteine, the wild types also exhibited copper tolerance. Moreover,Saccharomyces cerevisiae that expressedAtPCS1 showed tolerance to Cd but hypersensitivity to Cu. All these results indicate that PC is not a major factor in determining copper tolerance in plants.     

The isochorismate pathway is negatively regulated by salicylic acid signaling in O<Subscript>3</Subscript>-exposed <Emphasis Type="Italic">Arabidopsis</Emphasis>

Phytochelatins (PCs) are heavy metal binding peptides that play an important role in sequestration and detoxification of heavy metals in plants. In this study, our goal was to develop transgenic plants with increased tolerance for and accumulation of heavy metals from soil by expressing an Arabidopsis thaliana AtPCS1 gene, encoding phytochelatin synthase (PCS), in Indian mustard (Brassica juncea L.). A 35S promoter fused to a FLAG–tagged AtPCS1 cDNA was expressed in Indian mustard, and transgenic lines, designated pc lines, were evaluated for tolerance to and accumulation of Cd and Zn. Transgenic plants with moderate AtPCS1 expression levels showed significantly higher tolerance to Cd and Zn stress, but accumulated significantly less Cd and Zn than wild type plants in both shoot and root tissues. However, transgenic plants with highest expression of the transgene did not exhibit enhanced Cd and Zn tolerance. Shoots of Cd-treated pc plants had significantly higher levels of phytochelatins and thiols than wild-type plants. Significantly lower concentrations of gluthatione in Cd-treated shoot and root tissues of transgenic plants were observed. Moderate expression levels of phytochelatin synthase improved the ability of Indian mustard to tolerate certain levels of heavy metals, but at the same time did not increase the accumulation potential for Cd and Zn.     

Burkholderia sp. SCMS54 reduces cadmium toxicity and promotes growth in tomato

Cadmium (Cd) can enter soil through the use of fertilisers, calcareous, pesticides and industrial and/or domestic effluents. Cd can leach into groundwater and be taken up by plants, potentially leading to reductions in plant growth and yield. In soil, plant roots interact with heavy metal (HM)‐tolerant microorganisms that may promote plant growth. Soil microorganisms may also be able to solubilise or mobilise soil metals, thereby acting as bioremediators. A better understanding of the interaction among plants, metals, microorganisms and soil will lead to improved plant tolerance. Two multi‐tolerant bacteria from the Burkholderia genus were isolated from Cd‐contaminated and Cd‐uncontaminated soil of a coffee plantation. In addition to its high tolerance to Cd, the strain SCMS54 produces indole‐acetic acid (IAA), solubilises inorganic phosphate and produces siderophores, demonstrating its potential to contribute to beneficial plant–microorganism interactions. When interacting with tomato plants exposed to Cd, the bacterium led to decreases in plant peroxide and chlorosis levels, promoted relative plant growth and decreased the root absorption of Cd, resulting in increased plant tolerance to this highly toxic HM. The results indicated that the inoculation of tomato plants with Burkholderia sp. SCMS54 promotes better growth in plants cultivated in the presence of Cd. This phenomenon appears to be attributed to a mechanism that decreases Cd concentrations in the roots via a beneficial interaction between the bacteria and the plant roots.     

Phytochelatins and heavy metal tolerance

The induction and heavy metal binding properties of phytochelatins in heavy metal tolerant (Silene vulgaris) and sensitive (tomato) cell cultures, in water cultures of these plants and in Silene vulgaris grown on a medieval copper mining dump were investigated. Application of heavy metals to cell suspension cultures and whole plants of Silene vulgaris and tomato induces the formation of heavy metal–phytochelatin-complexes with Cu and Cd and the binding of Zn and Pb to lower molecular weight substances. The binding of heavy metal ions to phytochelatins seems to play only a transient role in the heavy metal detoxification, because the Cd- and Cu-complexes disappear in the roots of water cultures of Silene vulgaris between 7 and 14 days after heavy metal exposition. Free heavy metal ions were not detectable in the extracts of all investigated plants and cell cultures. Silene vulgaris plants grown under natural conditions on a mining dump synthesize low molecular weight heavy metal binding compounds only and show no complexation of heavy metal ions to phytochelatins. The induction of phytochelatins is a general answer of higher plants to heavy metal exposition, but only some of the heavy metal ions are able to form stable complexes with phytochelatins. The investigation of tolerant plants from the copper mining dump shows that phytochelatins are not responsible for the development of the heavy metal tolerant phenotypes.     

Transgenic Indian mustard (<Emphasis Type="Italic">Brassica juncea</Emphasis>) plants expressing an <Emphasis Type="Italic">Arabidopsis</Emphasis> phytochelatin synthase (<Emphasis Type="Italic">AtPCS1</Emphasis>) exhibit enhanced As and Cd tolerance

Phytochelatins (PCs) are post-translationally synthesized thiol reactive peptides that play important roles in detoxification of heavy metal and metalloids in plants and other living organisms. The overall goal of this study is to develop transgenic plants with increased tolerance for and accumulation of heavy metals and metalloids from soil by expressing an Arabidopsis thaliana AtPCS1 gene, encoding phytochelatin synthase (PCS), in Indian mustard (Brassica juncea L.). A FLAG-tagged AtPCS1 gDNA, under its native promoter, is expressed in Indian mustard, and transgenic pcs lines have been compared with wild-type plants for tolerance to and accumulation of cadmium (Cd) and arsenic (As). Compared to wild type plants, transgenic plants exhibit significantly higher tolerance to Cd and As. Shoots of Cd-treated pcs plants have significantly higher concentrations of PCs and thiols than those of wild-type plants. Shoots of wild-type plants accumulated significantly more Cd than those of transgenic plants, while accumulation of As in transgenic plants was similar to that in wild type plants. Although phytochelatin synthase improves the ability of Indian mustard to tolerate higher levels of the heavy metal Cd and the metalloid As, it does not increase the accumulation potential of these metals in the above ground tissues of Indian mustard plants.     

Wild plants, <Emphasis Type="Italic">Andropogon virginicus </Emphasis>L. and <Emphasis Type="Italic">Miscanthus sinensis</Emphasis> Anders,are tolerant to multiple stresses including aluminum,heavy metals and oxidative stresses

To isolate high-tolerant plants against aluminum (Al), heavy metals and/or oxidative stresses as a final goal, screening of Al tolerant plants from a collection of 49 wild plants was first of all performed in this study. Andropogon virginicus L. and Miscanthus sinensis Anders showed high Al tolerant phenotypes (more than 35% values in both relative root growth and germination frequency even under 900 μM Al concentration) in our screening. Al tolerance mechanisms in these two plants were characterized and the results suggested that (1) a transport system of toxic Al ions from root to shoot, (2) a suppression of Al accumulation in root tip region and (3) a suppression of oxidative damages by an induction of anti-peroxidation enzymes, such as superoxide dismutase (SOD) and catalase, were involved in the tolerance mechanisms. Six wild plants [Andropogon, Miscanthus, Dianthus japonicus Thunb, Echinochloa crus-galli (L.) Beauv, Reynoutria japonica Houtt, and Sporobolus fertilis (Steud.) W. Clayton] were furthermore tested for their sensitivity against heavy metal stresses and oxidative stresses. The two high Al tolerant plants, Andropogon and/or Miscanthus, showed tolerance to Cr, Zn, diamide or hydrogen peroxide, suggesting common tolerance mechanisms among the tested stresses. Reynoutria showed tolerance to diamide and hydrogen peroxide, Sporobolus to Cr and Echinocholoa to Cd and Cu. Moreover, the collection of wild plants used in this study was a very useful kit to isolate tolerant plants against various abiotic stresses within a short period of time.     

Mechanisms of cadmium detoxification in cattail (Typha angustifolia L.)

Cadmium (Cd) is a widespread heavy metal pollutant and environmental and human health hazard, which may be partially resolved using green and cost-effective phytoremediation techniques. However, the efficiency of phytoremediation is often limited by the small biomass of Cd-hyperaccumulator plants. Although cattail (Typha angustifolia L.) is tolerant of heavy metals and has a high biomass, there is little information available on its detoxification mechanisms for heavy metals, especially Cd. In the present study we investigated the tolerance of cattail to Cd and mechanisms involved in its Cd detoxification. Our results show that: (a) cattail is tolerant of Cd; (b) the root Casparian band, cell wall, vacuole, glutathione (GSH), and glutathione peroxidase (GPX) play important roles in Cd detoxification; and (c) mechanisms of Cd detoxification differ in leaf cell cytoplasm (mainly a GSH-related antioxidant defense system) and root cell cytoplasm (mainly a GSH-related chelation system). In summary, cattail possesses multiple detoxification mechanisms for Cd and is a promising species for phytoremediation of Cd-polluted environments.     

Root development and heavy metal phytoextraction efficiency: comparison of different plant species in the field

Heavy metal phytoextraction is a soil remediation technique which implies the optimal use of plants to remove contamination from soil. Plants must thus be tolerant to heavy metals, adapted to soil and climate characteristics and able to take up large amounts of heavy metals. Their roots must also fit the spatial distribution of pollution. Their different root systems allow plants to adapt to their environment and be more or less efficient in element uptake. To assess the impact of the root system on phytoextraction efficiency in the field, we have studied the uptake and root systems (root length and root size) of various high biomass plants (Brassica juncea, Nicotiana tabacum, Zea mays and Salix viminalis) and one hyperaccumulator (Thlaspi caerulescens) grown in a Zn, Cu and Cd contaminated soil and compared them with total heavy metal distribution in the soil. Changes from year to year have been studied for an annual (Zea mays) and a perennial plant (Salix viminalis) to assess the impact of the climate on root systems and the evolution of efficiency with time and growth. In spite of a small biomass, T. caerulescens was the most efficient plant for Cd and Zn removal because of very high concentrations in the shoots. The second most efficient were plants combining high metal concentrations and high biomass (willows for Cd and Zn and tobacco for Cu and Cd). A large cumulative root density/aboveground biomass ratio (L_A/B), together with a relative larger proportion of fine roots compared to other plants seemed to be additional favourable characteristics for increased heavy metal uptake by T. caerulescens. In general, for all plants correlations were found between L _A/B and heavy metal concentrations in shoots (r=0.758^***, r=0.594^***, r=0.798^*** (P<0.001) for Cd, Cu and Zn concentrations resp.). Differences between years were significant because of variations in climatic conditions for annual plants or because of growth for perennial plants. The plants exhibited also different root distributions along the soil profile: T. caerulescens had a shallow root system and was thus best suited for shallow contamination (0.2 m) whereas maize and willows were the most efficient in colonising the soil at depth and thus more applicable for deep contamination (0.7 m). In the field situation, no plant was able to fit the contamination properly due to heterogeneity in soil contamination. This points out to the importance and the difficulty of choosing plant species according to depth and heterogeneity of localisation of the pollution.     

Nodulation,biomass and nitrogen content in Sesbania species

Sesbania rostrata developed nitrogen fixing nodules on the stem after spraying the plants with the bacterial culture TCSR 1. The number of stem nodules at 55 days after sowing was about 1200. Plants with stem nodules had a significantly reduced number of root nodules. The biomass of S. rostrata was comparable to the locally well adapted non-stem nodulating species S. aculeata. The %N and total nitrogen content were highest in S. rostrata compared to the other three species studied.     

Heavy-metal uptake and distribution in Silene vulgaris and Minuartia verna growing on mining-dump material containing lead and zinc

Plant and soil samples from a mining area in Carinthia (Austria) were investigated for their heavy metal content. In the soil surrounding roots of plants (Minuartia verna and Silene vulgaris) growing on the mining dumps, high concentrations of lead and zinc are to be expected. The two species (Minuartia and Silene) show very different heavy metal concentrations in their above- and belowground organs. From these observations it can be concluded that the divergent distribution of heavy metals within the plants is an important mechanism of tolerance to heavy metals.     

铜、镉胁迫下外源NO介导的番茄解毒途径

一氧化氮(NO)作为信号分子,在抵御重金属胁迫中起重要作用,但对不同离子胁迫下的解毒机制尚缺乏研究.本研究采用营养液培养法,研究了铜(Cu)、镉(Cd)单一或复合胁迫下,番茄幼苗对Cu、Cd的吸收转运特性及对外源NO的响应机制.结果表明: 50 μmol·L^-1的Cu²⁺、Cd²⁺均显著抑制番茄植株的生长,其中Cd胁迫对生长的抑制效应远高于Cu胁迫.Cu、Cd单一或复合胁迫均使番茄根系Cu、Cd含量显著升高,但根系对Cu、Cd吸收存在严格选择性.根细胞对必需元素Cu表现出“奢侈吸收”的现象,而对毒性较强的Cd则吸收相对较少,胞内Cd浓度仅为Cu的1/10左右.外源NO处理可不同程度地缓解Cu、Cd胁迫,其中缓解Cd胁迫的效能更强.番茄对被动进入细胞的Cu、Cd具有相似的解毒机制:一方面,Cu、Cd胁迫诱导细胞质中产生谷胱甘肽(GSH)、植物螯合肽(PCs)和金属硫蛋白(MTs),络合过多的Cu、Cd离子,降低其生物毒性;另一方面,过多的Cu、Cd离子或螯合物被转运至液泡区隔化.外源NO通过调控GSH-GSSG(氧化型谷胱甘肽)氧化还原状态及GSH-PCs代谢方向的改变,促进Cu、Cd离子转运至液泡区隔化来缓解胁迫抑制;NO还可诱导植株叶片或根系表达更多的金属硫蛋白、GSH和PCs,而且上述响应普遍存在叠加效应.这可能是NO介导番茄对Cu、Cd胁迫的另一主要解毒途径.     

Factors affecting the distribution of cadmium,copper and lead and their effect upon yield and zinc content in bush beans (Phaseolus vulgaris L.)

Summary Concentrations of Cd, Pb and Cu in the roots, stems and leaves of bulgarian bush beans (Phaseolus vulgaris L.) were determined for plants grown in various soils of increasing levels of contamination of these metals. Most of each heavy metal absorbed by plants was retained in roots. Concentrations of Cd, Pb and Cu in roots increased in response to soil concentrations, whereas, in stems, only Cd and Pb concentrations increased and Cu concentration was relatively constant. It is thought that Cu transport to the stele was metabolically controlled, whereas Cd and Pb reached the stem by leakage across non suberised areas of the endodermis. Uptake of heavy metals was associated with a decrease in zinc content in plants and a decrease in yield. By regression analysis decrease in both zinc content and plant yield could be best related to Cd content in stems. Possible reasons for these effects are discussed.     

施用污泥对油菜根际养分和不同种类重金属的影响

根际是控制植物养分动态的重要因素,养分动态也影响着根际土壤环境。当土壤被污水污泥改良后,根际土壤中的养分和重金属性质也会发生变化。目前很少有人研究施用污泥的土壤中植物根系对根际重金属有效性和分布的影响。采用根垫—冰冻薄层切片法对施用污泥后土壤中油菜根际的养分和重金属分布情况进行研究,以期探明污泥改良土壤中根际重金属的活化特征。当土壤施用污泥后,根际土壤中DTPA提取态Zn,Cd,Ni,Mn,有效磷,有效钾和铵态氮被显著消耗,而根际土壤中DTPA提取态Cu没有明显的消耗或积累。当土壤中施用大量污泥时,根际土壤的pH值随着离根表面距离的增加而增加。无论土壤是否用污泥处理,油菜根际土壤中可交换态Cu都显著减少。当土壤被50%污泥改良时,在距离根表面0—2 mm处的油菜根际土壤中碳酸盐结合态,铁锰氧化物结合态,有机物结合态,残渣态的Cu和Zn都被消耗较多。污泥的施用对油菜的生长有促进作用。随着污泥施用量的增加,油菜地上部分Cu和Zn的含量没有显著变化。施用污泥量小于25%的土壤中,污泥没有增加重金属的可利用性和移动性。除了Cu,油菜根际土壤中DTPA提取态Zn,Cd,Ni的减少表明施用污泥的土壤中重金属的活化是非常有限的。     

Phytochelatins in Cadmium-Sensitive and Cadmium-Tolerant Silene vulgaris (Chain Length Distribution and Sulfide Incorporation)

In response to a range of Cd concentrations, the root tips of Cd-tolerant plants of Silene vulgaris exhibit a lower rate of PC production accompanied by a lower rate of longer chain PC synthesis than those of Cd-sensitive plants. At the same Cd exposure level, stable PC-Cd complexes are more rapidly formed in the roots of Cd-sensitive plants than in those of tolerant plants. At an equal PC concentration in the roots, the PC composition and the amount of sulfide incorporated per unit of PC-thiol is the same in both populations. Although these compounds might play some role in mechanisms that contribute to Cd detoxification, the ability to produce these compounds in greater amounts is not, itself, the mechanism that produces increased Cd tolerance in tolerant S. vulgaris plants.     

Histological observation of secondary aerenchyma formed immediately after flooding in Sesbania cannabina and S. rostrata

The effect of up to 48 h of flooding on the development of roots of Sesbania cannabina an0d S. rostrata seedlings was examined in a pot experiment. Light microscopy revealed that the outermost cells of the phellogen of the taproot of S. cannabina expanded and elongated during the first 12 h of flooding. After 18 h, the outermost of these regions was composed of cells that had expanded radially direction to form a spongy zone inside the endodermis. These elongated cells were radially connected to each other and formed the secondary aerenchyma surrounding the stele of taproot. While those histological alterations were not observed in S. rostrata, the number of layers of cells originating in the pericycle increased slightly, but elongation of the cells was not found during the first 18 h of flooding. After 36 h of flooding, cell elongation was also detected as outer layers of the phellogen. The delayed response to flooding in aerenchyma production in S. rostrata was compensated by immediate development of adventitious roots on submerged parts of the hypocotyl.     

Overexpression of phytochelatin synthase (<Emphasis Type="Italic">AtPCS</Emphasis>) in rice for tolerance to cadmium stress

Phytoremediation is an important strategy adapted by plants to sequester and/or detoxify pollutants. Phytochelatins, a family of cysteine-rich thiol-reactive peptides, bind to various heavy metals and metalloids making them good candidates for phytoremediation. Phytochelatin synthase catalyses the final step in the biosynthesis of phytochelatins and can be used as a strategy to improve tolerance against heavy metals. In the present study, an AtPCS gene was overexpressed in rice following the in planta transformation approach. Stringent screening strategies were standardized to select putative transformants under a Cd stress of 125 μM at both seedling and plant levels. Molecular analysis by PCR in 18 tolerant plants confirmed the transgene integration and absence of Agrobacterium. Genomic Southern analysis further confirmed the integration of the T-DNA as a single copy. The stability of the T-DNA in the progeny of 5 selected T₁ generation plants was confirmed by tolerance assay, molecular characterization and biochemical analysis for the reduced glutathione, phytochelatin content and lipid peroxidation. This strategy is discussed as a potential mechanism to enhance the tolerance of rice plants to Cd stress.     

Effects of Different Wetland Plant Species on Fresh Unweathered Sulphidic Mine Tailings

Vegetation cover with two Eriophorum species on old unweathered sulphidic mine tailings has earlier been found to reduce the element levels and to prevent production of acidity in drainage water. The present study aims to find out if Carex rostrata Stokes, Eriophorum angustifolium Honck. and Phragmites australis (Cav.) Steud. had other effects on metal and As release in fresh unweathered sulphidic mine tailings, if the species showed different effects and if this depended on plant mechanisms such as O₂, carbonate or organic acid release. Plants were grown in pots with fresh sulphidic mine tailings for 13 months. Arsenic, Cd, Cu, Fe, Pb, Zn, pH, SO ₄ ²⁻ , alkalinity and organic acids in the drainage water as well as metals and As in roots and shoot and O₂ and redox potential in pore water were analysed. The tailings weathered slowly due to high buffering capacity thus no pH decrease was found and therefore similar buffering effects by plants as shown in the previous investigation could not be found. The plants increased the total release of metals and As from the tailings. The release did not depend on carbonate or organic acid release from plants. However, the Fe and As release was due to changed redox potential, caused by O₂ release, and high concentration of Fe and As was found in plant roots. Phragmites australis released more As and Fe but less Cd than E. angustifolium and C. rostrata which make P. australis not suitable for plant establishment on sulphidic mine tailings containing high levels of As. Plants did take up the elements and the lowest translocation of elements to the shoot was found in P. australis while the highest in E. angustifolium.