Effects of copper deficiency and copper toxicity on organogenesis and some physiological and biochemical responses of Scots pine (Pinus sylvestris L.) seedlings grown in hydroculture

The morphological, physiological, and biochemical parameters of 6-week-old seedlings of Scots pine (Pinus sylvestris L.) were studied under deficiency (1.2 nM) and chronic exposure to copper (0.32, 1, 2.5, 5, and 10 μM CuSO₄) in hydroculture. The deposit of copper in the seed allowed the seedlings to develop under copper deficiency without visible disruption of growth. The high sensitivity of Scots pine to the toxic effects of copper was shown, which manifested as a significant inhibition of growth and development. The loss of dominance of the main root and a strong inhibition of lateral root development pointed to a lack of adaptive reorganization of the root system architecture under copper excess. A preferential accumulation of copper in the root and a minor translocation in aerial organs confirmed that Scots pine belongs to a group of plants that exclude copper. Selective impairment in the absorption of manganese was discovered, under both deficiency and excess of copper in the nutrient solution, which was independent of the degree of development of the root system. Following 10 μM CuSO₄ exposure, the absorption of manganese and iron from the nutrient solution was completely suppressed, and the development of seedlings was secured by the stock of these micronutrients in the seed. The absence of signs of oxidative stress in the seedling organs was shown under deficiency and excess of copper, as evidenced by the steady content of malondialdehyde and 4-hydroxyalkenals. Against this background, no changes in total superoxide dismutase activity in the organs of seedlings were revealed, and the increased content of low-molecular-weight antioxidants was observed in the roots under 1 μM and in the needles under 5 μM CuSO₄ exposures.

     

Laser cleaning of historical limestone buildings in Bordeaux appraisal using cathodoluminescence and electron paramagnetic resonance

BACKGROUND: Most historical buildings in Bordeaux city are made of limestone. This yellowish-white rock is rather porous and highly sensitive to pollution. As a consequence of local weathering conditions, these buildings present a dark appearance due to the development of a superficial dark grey to black crust. METHODS: For the last decade, a campaign has been underway to clean these buildings. Eleven techniques of surface treatment have been used, including laser beam technology. As a contribution to the study of laser beam effects on stone buildings, two analytical methods have been used on clean versus unclean surfaces: Cathodoluminescence (CL) and Electron Paramagnetic Resonance (EPR), in addition to SEM-EDX and XRD. RESULTS: The black crust is composed of different types of particles: carbon porous micro-particles of industrial origin, atmospheric dust due to the erosion of soils and rocks, alumino-silicate particles from urban pollution; all these particles being cemented by gypsum. DISCUSSION: As far as heritage conservation is concerned, the laser surface treatment not only preserves the original patina of the stone, but also leaves surface smoothness unaltered. CONCLUSIONS AND PERSPECTIVES: CL and EPR data confirm that lasers--with highly controlled parameters--only get rid of the black crust and, thus, reveal the underneath layer, the so-called patina. This patina shows no luminescence, whereas the limestone on which it has grown shows a bright orange emission of CL. This indicates CL to be a fast and easy way to provide a high quality control for the restoration of polluted ancient stones.     

Effects of copper on spore germination, growth and ultrastructure of Polypodium cambricum L. gametophytes

The effects of different concentrations (10(-5), 5x10(-5) and 10(-4)M) of copper bromide on spore germination, growth and ultrastructure were investigated in Polypodium cambricum L. gametophytes. The inhibitory effect of Cu was observed in spores cultured on medium supplemented with 10(-4)M CuBr(2): germination occurred about 40 days after sowing and was only 25%. Concentrations of 5x10(-5) and 10(-4)M CuBr(2) induced changes in gametophyte development, possibly by re-orientation of growth. Gametophytes treated with 10(-5) and 5x10(-5)M CuBr(2) took up and accumulated a large amount of copper and ultrastructural observations showed that cytoplasmic damage was limited to twisted swollen thylakoids. The ultrastructure of gametophytes treated with 10(-4)M CuBr(2) showed absence of a vacuolar compartment. The present observations suggest that P. cambricum gametophytes could be a suitable material for studying physiological and molecular alterations induced by excess copper.     

Evidence of singlet oxygen and hydroxyl radical formation in aqueous goethite suspension using spin-trapping electron paramagnetic resonance (EPR)

The generation of reactive species in an aqueous goethite suspension, under room light and aeration conditions, was investigated using the electron paramagnetic resonance (EPR) technique employing spin trap agents. The trap reagents, including 5,5-dimethylpyrroline N-oxide (DMPO) and 2,2,6,6-tetramethylpiperidine (TEMP), were used for the detection of OH radicals (OH) and singlet oxygen (¹O₂), respectively. On the addition of DMPO to the goethite suspended solution, a DMPO-OH adduct was formed, which was not decreased, even in the presence of the OH scavenger, mannitol. This result implied a false positive interpretation from the DMPO-OH EPR signal. In the presence of TEMP reagent, a TEMP-O signal was detected, which was completely inhibited in the presence of the singlet oxygen scavenger, sodium azide. With both DMPO-OH and TEMP-O radicals in the presence and absence of radical scavengers, singlet oxygen was observed to be the key species formed in the room-light sensitized goethite suspension. In the goethite/H₂O₂ system; however, both OH and singlet oxygen were generated, with significant portions of DMPO-OH resulting from both OH and singlet oxygen. In fact, the DMPO-OH resulting from OH should be carefully calculated by correcting for the amount of DMPO-OH due to singlet oxygen. This study reports, for the first time, that the goethite suspensions may also act as a natural sensitizer, such as fulvic acids, to form singlet oxygen.     

Catalytic reduction of NO on copper/MCM-41 studied by in situ EXAFS and XANES

Speciation of copper in the channels of MCM-41 during reduction of NO with CO at 473-773 K was studied by in situ extended X-ray absorption fine structural (EXAFS) and X-ray absorption near edge structural (XANES) spectroscopies in the present work. The component fitted (in situ) XANES spectra of the catalyst showed that about 72% of metallic copper (Cu(0)) in MCM-41 was oxidized to higher oxidation state coppers (Cu(II) (46%) and Cu(I) (26%)) during the NO reduction process (at 473 K). By EXAFS, we also found that in the NO reduction process, oxygen was inserted into the metallic copper matrix and led to a formation of the copper oxide species with a Cu-O bond distance of 1.93 A which was greater than that of the model compound Cu(2)O (typically 1.86 A). At 573-673 K, mainly Cu(II) was found in the channels of MCM-41. Nevertheless, at a higher temperature (e.g., 773 K), about 61% Cu(I), 31% Cu(II), and 8% Cu(O) with averaged Cu-Cu and Cu-O bond distances of 3.04 and 1.88 A, respectively were observed, that might account for the high selectivity-to-decomposition (S/D) ratios for yields of N(2) and CO(2) in the catalytic reduction of NO with CO.     

Effects of copper(II) and copper oxides on THMs formation in copper pipe

Little is known about how the growth of trihalomethanes (THMs) in drinking water is affected in copper pipe. The formation of THMs and chlorine consumption in copper pipe under stagnant flow conditions were investigated. Experiments for the same water held in glass bottles were performed for comparison. Results showed that although THMs levels firstly increased in the presence of chlorine in copper pipe, faster decay of chlorine as compared to the glass bottle affected the rate of THMs formation. The analysis of water phase was supplemented by surface analysis of corrosion scales using X-ray photoelectron spectroscopy (XPS), scanning electron microscope (SEM) and energy dispersive spectroscopy (EDX). The results showed the scales on the pipe surface mainly consisted of Cu₂O, CuO and Cu(OH)₂ or CuCO₃. Designed experiments confirmed that the fast depletion of chlorine in copper pipe was mainly due to effect of Cu₂O, CuO in corrosion scales on copper pipe. Although copper(II) and copper oxides showed effect on THMs formation, the rapid consumption of chlorine due to copper oxide made THM levels lower than that in glass bottles after 4 h. The transformations of CF, DCBM and CDBM to BF were accelerated in the presence of copper(II), cupric oxide and cuprous oxide. The effect of pH on THMs formation was influenced by effect of pH on corrosion of copper pipe. When pH was below 7, THMs levels in copper pipe was higher as compared to glass bottle, but lower when pH was above 7.     

Changes in the concentrations of phenolics and photosynthates in Scots pine (Pinus sylvestris L.) seedlings exposed to nickel and copper

Studies were done on the effects of elevated soil concentrations of copper (Cu) and (Ni) on foliar carbohydrates and phenolics in Scots pine (Pinus sylvestris L.). Four year-old seedlings were planted in pots filled with metal-treated mineral forest soil in early June. The experimental design included all combinations of four levels of Cu (0, 25, 40 and 50 mg kg(-1) soil dw) and Ni (0, 5, 15 and 25 mg kg(-1) soil dw). Current year needles were sampled for soluble sugar, starch and phenolics at the end of September. Ni increased sucrose concentration in the needles, indicating disturbances in carbohydrate metabolism. Trees exposed to Ni had higher concentrations of condensed tannins compared with controls. In contrast, concentrations of several other phenolic compounds decreased when seedlings were exposed to high levels of Cu or to a combination of Ni and Cu. The results suggest that concentrations of phenolics in Scots pine needles vary in their responses to Ni and Cu in the forest soil.     

Effects of photochemical smog and mineral nutrition on ponderosa pine seedlings

Two-year-old seedlings of ponderosa pine (Pinus ponderosa Dougl. ex Laws) were exposed to ambient concentrations of photochemical smog (AA) and clean air (CA) during a single field season at Tanbark Flat of the San Gabriel Mountains in the Los Angeles Basin. The seedlings were grown in a perlite-vermiculite medium with full supply of nutrients (based on modified Hoagland solution); reduced to 50% supply of N; reduced to 50% supply of Mg; and reduced to 50% supply of N+Mg. No significant effects of air pollution exposures on injury development, stem growth and concentrations of plant pigments were determined. The seedlings in the AA treatment had decreased N concentration in current year needles compared with CA seedlings; however, the needle concentrations of other elements did not change. Reduction of N supply in the growing medium caused decreased N, P, Ca, K and chlorophyll a concentrations in needles. Stem growth of the seedlings with reduced N supply was significantly decreased as well. No changes in stem growth or chemical composition of plants with reduced Mg supply were noted. Reduction of supply of nutrients did not change responses of trees to the air pollution exposures.     

4种重金属离子对间接火焰原子吸收光谱法测定铝的干扰

空气-乙炔间接火焰原子吸收法是我国目前运用较多且易于推广的铝检测方法,研究了Mg2+、Ca2+、Zn2+和Ni2+4种离子对此法测定0.40 mg/L Al3+ 的干扰情况。得出如下结论:(1)在±5% 误差范围内,15.10 mg/L Mg2+、105.00 mg/L Ca2+、0.23 mg/L Zn2+、0.88 mg/L Ni2+对0.40 mg/L Al3+ 无干扰。(2)本方法虽然操作复杂,但成本较低、灵敏度高,检测的浓度范围宽、稳定性好,加标回收率为96%~99%,检出限为0.04 mg/L,适宜测定范围为0.05~100.00 mg/L,适用于饮用水、河水及地下水中Al3+ 的测定。     

外源有机酸对小飞扬草(Euphorbia thymifolia L.)修复镉污染土壤的影响

植物修复有许多优点,但当用于重金属含量较高的土壤时,通常修复效率较低。研究了添加外源柠檬酸、草酸、苹果酸对镉超积累植物小飞扬草(Euphorbia thymifolia L.)富集Cd的影响,以及3种外源有机酸对土壤pH、Cd形态及小飞扬草根系生理生化特性的影响,探讨利用有机酸强化小飞扬草修复镉污染土壤的可能性及其作用机制。结果表明,土壤Cd含量为247 mg/kg时,种植15 d后,3种有机酸均提高了小飞扬草根和地上部Cd含量,强化效果:草酸>柠檬酸>苹果酸,10 mmol/kg草酸的加入使小飞扬草地上部分Cd含量达最大值77.21 mg/kg,比对照组提高87.72%。加入有机酸使土壤pH从7.65下降至6.01~7.45,BCR三步提取法分析土壤Cd形态结果表明,酸溶态Cd含量增加,残渣态Cd含量减小。加入20 mmol/kg苹果酸降低了根系的活力和ATP酶活性,根细胞膜透性与对照组无显著性差异,而加入5、10和20 mmol/kg的柠檬酸、草酸和5、10 mmol/kg的苹果酸均显著增加了小飞扬草根系的活力和ATP酶活性,减小了根细胞膜透性。说明添加3种外源有机酸能强化小飞扬草修复Cd污染土壤,利用外源有机酸很有可能成为提高重金属污染土壤植物修复效率的有效途径。     

Zinc and copper uptake by plants under two transpiration rates. Part II. Buckwheat (Fagopyrum esculentum L.)

To evaluate the environmental risks of irrigating crops with treated wastewater, a study was undertaken to quantify heavy metal uptake by 4-week old buckwheat (Fagopyrum esculentum L.) plants during 18 days of irrigation with 8 different Cu and Zn solutions under two transpiration rates (TR). At 4 weeks, potted buckwheat plants were transferred into one of the two growth chambers, offering either a high or low vapour pressure deficit (VDP) for, respectively, a high or low TR. Triplicate pots received one of the 8 irrigation treatments containing one of two Zn levels (0 and 25 mg/L) combined with one of four Cu levels (0, 5, 10 and 15 mg/L). Daily TR were measured by weighing the evapo-transpired water lost from the planted pot, less was the evaporation loss measured from triplicate non-planted pots. After 0, 6, 12 and 18 days of treatment, the stems and leaves of three randomly selected plants were harvested and after 18 days, the roots were harvested to determine Cu and Zn uptake. The treatments did not affect TR in terms of dry plant mass, indicating the absence of toxic effects. Irrigating with Zn, without Cu, increased dry biomass production, whereas the lowest biomass occurred with 15 and 30 mg/L of Cu with and without 25 mg/L of Zn, respectively, because higher applications of heavy metal significantly reduced soil pH. Plant Cu and Zn uptake increased with TR. With higher levels of Cu, Zn uptake by buckwheat was significantly reduced, while Zn had a slight but non-significant impact on Cu uptake. Previously and in a study exposing wheat plants to the same conditions, Cu significantly increased Zn uptake, while Zn had a slight but insignificant negative effect on Cu uptake. The buckwheat roots contained the greatest levels of Cu and Zn, indicating their role in moderating heavy metal uptake. Also, both Cu and Zn had a synergetic effect on each other in terms of root levels, and a similar observation was made in the earlier similar experiment using wheat plants. Irrigating a buckwheat crop with treated wastewater, with more natural Cu and Zn levels of 0.08 mg/L, could be quite beneficial without endangering the quality of the crop and acidifying the soil pH. The most concentrated experimental solutions contained 300 times more Cu and Zn, to obtain measurable differences.     

Zinc and copper uptake by plants under two transpiration rates. Part I. Wheat (Triticum aestivum L.)

To evaluate the environmental risk of irrigating crops with treated wastewater, an experiment was conducted using two growth chambers, each offering a different vapour pressure deficit (VPD) for high and low transpiration rates (TR), respectively. One of the two sets of 24 pots planted with 6 week old wheat (Triticum aestivum L.), was placed in each growth chamber, and irrigated in triplicates for 20 days with 8 Zn and Cu solutions (0 and 25 mg Zn/L combined with 0, 5, 15 and 30 mg Cu/L). Water losses from planted and non-planted pots served to measure evapo-transpiration and evaporation, respectively. Pots were monitored for Cu and Zn uptake by collecting three plants (shoot and grain)/pots after 0, 10 and 20 days, and roots in each pot after 20 days, and analyzing these plant parts for dry mass, and Cu and Zn levels. Transpiration rate was not affected by any Cu/Zn treatment, but Cu and Zn uptake increase with the time, irrigation solution level and higher TR, with the roots retaining most Cu and Zn, compared to the shoot followed by the grain. For the shoot and grain, Cu had a significant synergetic effect on Zn uptake, when Zn had slight but insignificant antagonistic effects on Cu uptake. For the roots, Cu and Zn had significant synergetic effect on each other. Regression equations obtained from the data indicate that Cu and Zn levels normally found in treated wastewater (0.08 mg/L) are 300 times lower than those used for the most concentrated experimental solutions (30 and 25 mg/L, respectively) and may, on a long term basis, be beneficial rather than toxic to wheat plants and do not acidify soil pH.     

Effects of silicon and copper on bamboo grown hydroponically

Due to its high growth rate and biomass production, bamboo has recently been proven to be useful in wastewater treatment. Bamboo accumulates high silicon (Si) levels in its tissues, which may improve its development and tolerance to metal toxicity. This study investigates the effect of Si supplementation on bamboo growth and copper (Cu) sensitivity. An 8-month hydroponic culture of bamboo Gigantocloa sp. “Malay Dwarf ” was performed. The bamboo plants were first submitted to a range of Si supplementation (0–1.5 mM). After 6 months, a potentially toxic Cu concentration of 1.5 μM Cu²⁺ was added. Contrary to many studies on other plants, bamboo growth did not depend on Si levels even though it absorbed Si up to 218 mg g^?1 in leaves. The absorption of Cu by bamboo plants was not altered by the Si supplementation; Cu accumulated mainly in roots (131 mg kg^?1), but was also found in leaves (16.6 mg kg^?1) and stems (9.8 mg kg^?1). Copper addition did not induce any toxicity symptoms. The different Cu and Si absorption mechanisms may partially explain why Si did not influence Cu repartition and concentration in bamboo. Given the high biomass and its absorption capacity, bamboo could potentially tolerate and accumulate high Cu concentrations making this plant useful for wastewater treatment.     

Effects of ammonium and aluminium on the development and nutrition of Pinus nigra in hydroculture

Application of ammonium and aluminium to young Pinus nigra var. maritima (Ait.) Melville trees resulted in a variety of negative effects. Excess ammonium led to an increase in shoot/root ratio. The biomass of the fine roots declined, resulting in an increase of the coarse/fine root ratio. The degree of mycorrhizal infection of the roots decreased. The nitrogen content of the trees increased considerably, whereas particularly the levels of calcium magnesium, manganese and zinc decreased sharply. Excess aluminium resulted in a simultaneous reduction of root and shoot biomass, a decline of the fine root system, an increase in the coarse/fine root ratio and a decrease in the degree of mycorrhizal infection. Uptake of the divalent cations calcium, magnesium, iron, manganese and zinc was restricted substantially, The nitrogen and phosphorus contents of the trees were hardly affected, whilst the potassium content of the shoot increased and of the roots decreased. This implicates that a deteriorating fine root system has to supply water and nutrients to a more demanding shoot. In the long term, high ammonium inputs and aluminium dissolution in forest ecosystems will lead to substantial nutrient deficiencies, just as has been found in the field.     

Effects of copper and palladium on the reduction of bromate by Fe(0)

Bromate reduction by Fe(0) with incorporation of copper or palladium was investigated in batch tests. The incorporation of copper led to an increase in the rate of bromate reduction, while incorporation of palladium did not show any effect on bromate reduction by Fe(0), regardless of the bimetal application techniques (either simultaneous addition of Cu(II) or Pd(IV) into the Fe-BrO3- reaction system or using copper or palladium amended iron for bromate removal). Surface analyses by X-ray photoelectron spectroscopy (XPS) and X-ray powder diffraction (XRD) techniques indicated that aqueous Cu(II) was reduced and incorporated into the iron surface to form Cu2O and Cu(0). Among these two species, pure Cu(0) is not an active electron donor to the bromate reduction reaction, as shown by there being no reduction from using Cu(0) powders alone and no enhancement by Fe(0) when physically mixed with Cu(0). Although it has been proposed in the literature that the enhancement of adsorption also contributes to the enhancement of chemical reduction, this is not the case here because adsorption decreased when Cu increased. The enhanced bromate reduction rate in the presence of copper observed here is most likely the result of the newly formed active Cu(I). The presence of PdO was evidenced by XPS but yielded no enhancement in bromate reduction. Finally, the Cu2O present on the iron surface because of copper impurities in commercially available iron was found to be involved in the bromate reduction and to accelerate the reduction rate.     

Effects of root anatomy and Fe plaque on arsenic uptake by rice seedlings grown in solution culture

Hydroponic experiments were carried out to investigate the effects of root anatomy, induced by aeration and stagnation, and Fe plaque on arsenic (III&V) uptake and translocation by rice plants. The results showed that As uptake in rice plants (Gui Chao-2) treated by aeration was decreased due to lower root specific surface area. Rice roots with larger specific surface area tended to form more Fe plaque, and Fe plaque affected As uptake kinetics by changing As influx curves from linear to hyperbolic for As(III) and from hyperbolic to S-curve for As(V). Fe plaque increased As(III&V) adsorption and minimized the effects of root anatomy characteristics on As uptake into roots and subsequently translocation to shoots. Fe plaque increased As(III) uptake rate at As(III) concentrations of 0.5∼8 mg L⁻¹, reduced As(V) uptake rate at low As(V) concentrations (<2 mg L⁻¹), but increased As uptake rate at high As(V) concentrations (>6 mg L⁻¹).