A two-compartment exposure device for foliar uptake study

An airtight two-chamber exposure devise was designed for investigating foliar uptake of polycyclic aromatic hydrocarbons (PAHs) by plants. The upper and the bottom chambers of the device were air-tightly separated by an aluminum foil and the plant aerial tissues and roots were exposed in the two chambers, respectively. The device was tested using maize exposed to several PAH species. Positive correlations between air and aerial tissue concentrations of the exposed PAH species were revealed. PAHs spiking in the culture solution had no influence on the leaf concentrations.     

Accumulation and distribution of polycyclic aromatic hydrocarbons in rice (Oryza sativa)

Various tissues of rice plants were sampled from a PAH contaminated site in Tianjin, China at different growth stages of the ripening period and analyzed for PAHs. PAHs were much higher in roots than in the exposed tissues. Grains and internodes accumulated much smaller amounts of PAHs than leaves, hulls or ear axes. No specific gradient trends along roots, stem, ear axes, and grains were observed, suggesting that systematic translocation among them is unlikely. Over the ripening period, PAH concentrations were increased in rice roots and decreased in most above-ground tissues. Significant correlations between PAH and lipid contents can only be observed during full mature stage. The spectra of individual PAH compounds in rice organs including roots were similar to those in air, rather than those in soil. There was also a significant correlation between bioconcentration factor (BCF, plant over air) and octanol/air partitioning coefficient (Koa).     

Polycyclic aromatic hydrocarbons in leaf cuticles and inner tissues of six species of trees in urban Beijing

Leaf samples of six tree species were collected along urban roadsides and a campus site in Beijing for measurement of polycyclic aromatic hydrocarbons (PAHs). PAHs in leaves were attributed to two fractions, leaf cuticles and inner leaf tissues, using sequential extraction. Total concentrations of 16 PAHs in the cuticles and the inner tissues were 69.3+/-64.6 microg g(-1) (d.w.) and 1.07+/-0.2 microg g(-1) (d.w.) at roadside and 57.5+/-52.6 microg g(-1) and 0.716+/-0.2 microg g(-1) on campus, respectively. The lipid-normalized inner tissue PAHs varied from 5.8 microg g(-1) to 15.0 microg g(-1). Similarities in PAH spectra between leaf cuticles and airborne particles and between the inner tissues and gaseous phase imply that airborne particulates and gaseous PAHs are likely the sources of PAHs for cuticles and the inner tissues, respectively. Difficulty in migration of heavier PAHs into inner tissues could be another reason.     

Bioconcentration of polycyclic aromatic hydrocarbons in vegetables grown in an industrial area

The polycyclic aromatic hydrocarbon (PAH) content was determined in the inner tissue of various vegetable species and their growing environment (soil and atmosphere) in the greater industrial area of Thessaloniki, northern Greece. The lower molecular weight compounds dominated in both vegetable leaves and roots. Statistical analysis of variance showed that species and season are the factors that significantly affect PAH concentrations in inner vegetable tissue and soil, respectively. Principal component analysis indicated that the mixture of PAHs in inner vegetable tissue was very similar to that in air vapour thus suggesting gaseous deposition as the principal pathway for the accumulation of PAHs. Soil-to-vegetation and air-to-vegetation bioconcentration factors were calculated and their relationships with PAHs' physicochemical properties were investigated. Solubility and the octanol-water partition coefficient, as well as vapour pressure and the octanol-air partition coefficient were proved to be good predictors for the accumulation of PAHs in inner root and leaf tissue, respectively.     

Distribution of PAHs in tissues of wetland plants and the surrounding sediments in the Chongming wetland, Shanghai, China

Polycyclic aromatic hydrocarbons (PAHs) concentrations were determined in sediments and three types of wetland plants collected from the intertidal flats in the Chongming wetland. The concentration of total PAHs in sediments ranged from 38.7 to 136.2 ng g⁻¹. Surface sediment concentrations were higher in regions with plant cover than in bare regions. Rhizome-layer sediments (56.8-102.4 ng g⁻¹) contained less PAHs than surface sediments (0-5 cm). Concentrations of PAHs in plant tissues ranged from 51.9 to 181.2 ng g⁻¹, with highest concentrations in the leaves of Scirpus. Most of the PAHs in the leaves and other plant tissues were low molecular weight compounds (LMW, 2-4 rings), and a similar distribution pattern of PAHs in different types of plants was also observed. Source analysis indicated that plants and sediments both came from pyrogenic sources, but plants had additional petroleum contamination. The low ratio of benzo[a]anthracene over chrysene suggests that the wetland PAHs came mainly from long-distance atmospheric transportation. Significant bioaccumulation of PAHs from the sediments into plants was not observed for high molecular weight PAHs (HMW, 5-6 rings) in Chongming wetland. The small RCFs (root concentration factor from sediments) for HMW PAHs and large RCFs for LMW PAHs suggested that roots accumulated LMW PAHs selectively from sediments in Chongming wetland.     

Allocation and source attribution of lead and cadmium in maize (Zea mays L.) impacted by smelting emissions

Plants grown in contaminated areas may accumulate trace metals to a toxic level via their roots and/or leaves. In the present study, we investigated the distribution and sources of Pb and Cd in maize plants (Zea mays L.) grown in a typical zinc smelting impacted area of southwestern China. Results showed that the smelting activities caused significantly elevated concentrations of Pb and Cd in the surrounding soils and maize plants. Pb isotope data revealed that the foliar uptake of atmospheric Pb was the dominant pathway for Pb to the leaf and grain tissues of maize, while Pb in the stalk and root tissues was mainly derived from root uptake. The ratio of Pb to Cd concentrations in the plants indicated that Cd had a different behavior from Pb, with most Cd in the maize plants coming from the soil via root uptake.     

Metal accumulation in aquatic macrophytes from southeast Queensland,Australia

To determine the extent of metal accumulation in some aquatic macrophytes from contaminated urban streams in southeast Queensland, plants were sampled from six sites, along with contiguous sediments. In all, 15 different species were collected, the most common genera being Typha (Cattails or Bulrushes) and Persicaria (Knotweeds). Before heavy metal analysis, plants were further separated into various morphological tissues, and five selected samples were separated into various physiological tissues. The cadmium, copper, lead and zinc content of the plants were analysed using flames AAS. In general, plant roots exhibited higher metal concentrations than the contiguous sediments. Of the metals of interest, only for zinc was there a relatively clear pattern of increasing accumulation in aquatic macrophytes with increasing sediment metal concentrations. Comparison between morphological tissues of the sampled plants found that roots consistently presented higher metal concentrations than either the stems or leaves, however unlike previous studies, this investigation revealed no consistent trend of stems accumulating more metals than the leaves. For Typha spp., metal concentrations followed the order of roots > rhizomes > leaves, while for Persicaria spp. the order was roots > leaves > stems. The submerged species Myriophyllum aquaticum accumulated the highest levels of metals overall (e.g. Zn 4300 micrograms g-1 dry weight and Cd 6.5 micrograms g-1), and the emergent macrophytes also exhibited relatively high metal contents in their roots. The leaves of the submerged and floating-leafed species collected contained relatively high quantities of the four metals of interest, compared with the leaves of emergent aquatic macrophytes. In the Typha rhizome and Persicaria stem samples analysed for internal variation in metal content, there was a pattern of increasing metal concentrations towards the external sections of the stem, both for subterranean stems (rhizomes) and above-substrate stems. For Persicaria stems, no clear pattern was observed for cadmium and lead, the two metals investigated that are not required by plants for survival.     

Evidence for bioaccumulation of PAHs within internal shoot tissues by a halophytic plant artificially exposed to petroleum-polluted sediments

Polycyclic aromatic hydrocarbons (PAHs) are widespread environmental pollutants of natural and anthropic origins. Despite their poor water solubility, they can be taken up and bioaccumulated by plants. This study was aimed at determining whether the PAHs present in sediments artificially polluted by heavy fuel oil are transferred to shoots of a coastal and edible plant, Salicornia fragilis Ball et Tutin. Bioaccumulation was quantified after a one-week exposure to sediments polluted with 0.2%, 2% and 20% fuel oil (w/w) and over a six-week monitoring at 0.2%. Quantification by GC-MS of PAH amounts in plants and sediments evidenced a bioaccumulation in the shoots by a soil-to-plant transfer through the root system. This bioaccumulation depended on the duration of exposure and on the substratum contamination. PAHs distributions in plants and sediments looked alike with a predominance of low- and medium-weight hydrocarbons. Moreover, high-weight PAHs were also detected in the upper part of plants.     

Agricultural reuse of reclaimed water and uptake of organic compounds: pilot study at Mutah University wastewater treatment plant, Jordan

The residues of polynuclear aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), chlorinated benzenes (CBs) and phenols were investigated for soil, wastewater, groundwater and plants. The uptake concentration of these compounds was comparatively determined using various plant types: Zea mays L., Helianthus annus L., Capsicum annum L., Abelmoschus esculentus L., Solanum melongena L. and Lycopersicon esculentum L. which were grown in a pilot site established at Mutah University wastewater treatment plant, Jordan. Soil, wastewater, groundwater and various plant parts (roots, leaves and fruits) samples were extracted in duplicate, cleaned up by open-column chromatography and analyzed by a multi-residue analytical methods using gas chromatography equipped with either mass selective detector (GC/MS), electron capture detector (GC/ECD), or flame ionization detector (FID). Environmentally relevant concentrations of targeted compounds were detected for wastewater much higher than for groundwater. The overall distribution profiles of PAHs and PCBs appeared similar for groundwater and wastewater indicating common potential pollution sources. The concentrations of PAHs, PCBs and phenols for different soils ranged from 169.34 to 673.20 microg kg(-1), 0.04 to 73.86 microg kg(-1) and 73.83 to 8724.42 microg kg(-1), respectively. However, much lower concentrations were detected for reference soil. CBs were detected in very low concentrations. Furthermore, it was found that different plants have different uptake and translocation behavior. As a consequence, there are some difficulties in evaluating the translocation of PAHs, CBs, PCBs and phenols from soil-roots-plant system. The uptake concentrations of various compounds from soil, in which plants grown, were dependent on plant variety and plant part, and they showed different uptake concentrations. Among the different plant parts, roots were found to be the most contaminated and fruits the least contaminated.     

Chromium induced lipid peroxidation in the plants of Pistia stratiotes L.: role of antioxidants and antioxidant enzymes

In the plant, Pistia stratiotes L., the effect of different concentrations of chromium (0, 10, 40, 80 and 160 microM) applied for 48, 96 and 144 h was assessed by measuring changes in the chlorophyll, protein, malondialdehyde (MDA), cysteine, non-protein thiol, ascorbic acid contents and superoxide dismutase (SOD), ascorbate peroxidase (APX) and guiacol peroxidase (GPX) activity of the plants. Both in roots and leaves, an increase in MDA content was observed with increase in metal concentration and exposure periods. In roots, the activity of antioxidant enzymes viz. SOD and APX increased at all the concentrations of Cr at 144 h than their controls. The GPX activity of the treated roots increased with increase in Cr concentration at 48 and 96 h of exposures, however, at 144 h its activity was found declined beyond 10 microM Cr. The level of antioxidants in the roots of the treated plant viz. cysteine and ascorbic acid was also found increased at all the concentrations of Cr at 144 h than their respective controls, however, an increase in the non-protein thiol content was recorded up to 40 microM Cr followed by decrease. The chlorophyll content decreased with increase in Cr concentrations and exposure periods. However, the protein content of both roots and leaves were found decreased with increase in Cr concentrations at all the exposure periods except an increase was recorded at 10 microM Cr at 48 h. In Cr treated plants, the no observed effect concentration (NOEC) and lowest observed effect concentration (LOEC) for leaves chlorophyll and protein contents were 40 and 80 microM Cr, respectively after 48 h exposure while NOEC and LOEC for root protein content were 10 and 40 microM, respectively after 48 h. The analysis of correlation coefficient data revealed that the metal accumulation in the roots of the plant was found positively correlated with antioxidant parameters except SOD after 48 h of exposure, however, negatively correlated with most of all the parameters studied at 144 h in both part of the plant. It may be suggested from the present study that toxic concentrations of Cr cause oxidative damage as evidenced by increased lipid peroxidation and decreased chlorophyll and protein contents. However, the higher levels of enzymatic and non-enzymatic antioxidants suggest the reason for tolerating higher levels of metals.     

Influence of cadmium on antioxidant capacity and four microelement concentrations in tomato seedlings (Lycopersicon esculentum)

Tomato (Lycopersicon esculentum) seedlings were grown in four cadmium (Cd) levels of 0-10 microM in a hydroponic system to analyze the antioxidative enzymes, Cd concentration in the plants, and the interaction between Cd and four microelements. The results showed that there was a significant increase in malondialdehyde (MDA) concentration, and superoxide dismutase (SOD) and peroxidase (POD) activities in the plants subjected to 1-10 microM Cd. This indicates that Cd stress induces an oxidative stress response in tomato plants, characterized by an accumulation of MDA and increase in activities of SOD and POD. Root, stem and leaf Cd concentrations increased with its exposure Cd level, and the highest Cd concentration occurred in roots, followed by leaves and stems. A concentration- and tissue-dependent response was found in the four microelement concentrations to Cd stress in the tomato leaves, stems and roots. Regression analysis showed that there was a significantly negative correlation between Cd and Mn, implying the antagonistic effect of Cd on Mn absorption and translocation. The correlation between Cd and Zn, Cu and Fe were inconsistent among leaves, stems and roots.     

Atmospheric phenanthrene pollution modulates carbon allocation in red clover (Trifolium pratense L.)

The influence of atmospheric phenanthrene (PHE) exposure (160 μg m⁻³) during one month on carbon allocation in clover was investigated by integrative (plant growth analysis) and instantaneous ¹³CO₂ pulse-labelling approaches. PHE exposure diminished plant growth parameters (relative growth rate and net assimilation rate) and disturbed photosynthesis (carbon assimilation rate and chlorophyll content), leading to a 25% decrease in clover biomass. The root-shoot ratio was significantly enhanced (from 0.32 to 0.44). Photosynthates were identically allocated to leaves while less allocated to stems and roots. PHE exposure had a significant overall effect on the ¹³C partitioning among clover organs as more carbon was retained in leaves at the expense of roots and stems. The findings indicate that PHE decreases root exudation or transfer to symbionts and in leaves, retains carbon in a non-structural form diverting photosynthates away from growth and respiration (emergence of an additional C loss process).     

Polynuclear aromatic hydrocarbons in crops from long-term field experiments amended with sewage sludge

This study reports on the polynuclear aromatic hydrocarbon (PAH) content of crop plants grown at three different field experiments where controlled additions of sewage sludge were made a number of years ago. Archived samples of several crops have been analysed from sludge-amended and unsludged control plots for 15 PAH compounds. Root crops and above ground plant parts were available for several years following the last applications of sludge. Although the soil PAH burden increased substantially due to sewage sludge additions, and residues of these compounds have persisted in the soils for many years, increased PAH concentrations relative to the unsludged controls were not consistently detected in plant tissues. Plant samples were relatively enriched with low molecular weight compounds such as acenaphthene/fluorene nd phenanthrene. Various lines of evidence indicate that PAHs detected in above ground plant parts are chiefly derived from atmospheric inputs, while PAHs detected in root crops probably arise from adsorption to the root surface.     

Heavy metal accumulation in Halimione portulacoides: intra- and extra-cellular metal binding sites

Salt marsh plants can sequestrate and inherently tolerate high metal concentrations found in salt marsh sediments. This work intended to understand the Halimione portulacoides (L.) Aellen strategies to prevent metal toxicity, by investigating the metal location in different plant organs and in the cell. A sequential extraction was performed on leaves, stems and roots of H. portulacoides in order to determine and compare the metal (Zn, Pb, Co, Cd, Ni and Cu) concentration in several fractions of the plant material (ethanolic, aqueous, proteic, pectic, polissacaridic, lenhinic and cellulosic). This study shows that all plant organs of H. portulacoides mostly retain metals in the cell wall (65% is the average for all studied metals stored in the root cell wall, 55% in the stems and 53% in the leaves), and the metal content in the intracellular compartment is much lower (21% in roots, 25% in stems and 32% in leaves). High levels of heavy metal in the sedimentary environment do not cause toxicity to H. portulacoides, because H. portulacoides immobilizes them in different cell compartments (cell wall+proteic fraction+intracellular) outside key metabolic sites.     

Molecular evidence for benzo[a]pyrene and naphthalene genotoxicity in Trifolium repens L

Polycyclic aromatic hydrocarbons (PAHs) are among the most dangerous environmental contaminants due to their toxic, carcinogenic and mutagenic effects. Although there are many data in literature that detail the effects of PAHs on animals, little is known about their action on higher plants which are often used as bioindicators. The aim of the present study was to evaluate the genotoxicity of two different PAHs, benzo[a]pyrene (BaP) and naphthalene (Naph), on Trifolium repens L. Clover plants were exposed to soil which had been artificially contaminated with three concentrations of BaP (5, 10 and 20 microg g-1) or Naph (25, 50 and 100 microg g-1). After 15 days, changes in the DNA content and sequence of roots and shoots were evaluated by flow cytometry (FCM) and amplified fragment length polymorphism (AFLP). Root and shoot dry weight were also determined to assess plant growth. Results showed that BaP and Naph were both genotoxic for white clover, inducing significant changes in root and shoot DNA sequence. Damage was more severe in the root than in the shoot suggesting that the translocation of these compounds and their genotoxic metabolites was limited. Ploidy alterations were not detected and the extent of damage caused by all the tested PAH concentrations was not sufficient to affect plant development.     

Biological responses of wheat (Triticum aestivum) plants to the herbicide chlorotoluron in soils

Chlorotoluron is a phenylurea herbicide that is widely used for controlling grass weeds in the land of cereal, cotton and fruit production. However, extensive use of this herbicide may lead to its accumulation in ecosystems, thus inducing the toxicity to crops and vegetables. To assess chlorotoluron-induced toxicity in plants, we performed the experiment focusing on the metabolic adaptation of wheat plants (Triticum aestivum) to the chlorotoluron-induced oxidative stress. The wheat plants were cultured in the soils with chlorotoluron at concentrations of 0-25mg/kg. Chlorotoluron accumulation in plants was positively correlated with the external chlorotoluron concentrations, but negatively with the plant growth. Treatment with chlorotoluron induced the accumulation of O(2)(-) and H(2)O(2) in leaves and resulted in the peroxidation of plasma membrane lipids in the plant. We measured the endogenous proline level and found that it accumulated significantly in chlorotoluron-exposed roots and leaves. To understand the biochemical responses to the herbicide, activities of the antioxidant enzymes, such as superoxide dismutase (SOD), peroxidase (POD), catalase (CAT) and ascorbate peroxidase (APX) were assayed. Analysis of SOD activity by non-denaturing polyacrylamide gel electrophoresis (PAGE) revealed that there were three isoforms in the roots and leaves, but the isoforms in the tissues showed different patterns. Also, using the native PAGE, 6 isoforms of root POD and 10 in leaves were detected. The total activity of POD in roots was significantly enhanced. Activities of APX in roots and leaves showed a similar pattern. The CAT activities were generally suppressed under the chlorotoluron exposure.     

Uptake of polycyclic aromatic hydrocarbons (PAHs) in salt marsh plants Spartina alterniflora grown in contaminated sediments

Polycyclic aromatic hydrocarbon (PAH) concentrations were measured in Spartina alterniflora plants grown in pots of contaminated sediment, plants grown in native sediment at a marsh contaminated with up to 900 microg/g total PAHs, and from plants grown in uncontaminated control sediment. The roots and leaves of the plants were separated, cleaned, and analyzed for PAHs. PAH compounds were detected at up to 43 microg/g dry weight in the root tissue of plants grown in pots of contaminated soil. PAH compounds were detected at up to 0.2 microg/g in the leaves of plants grown in pots of contaminated soil. Concentrations less than 0.004 microg/g were detected in the leaves of plants grown at a reference site. Root concentration factor (RCF) values ranged from 0.009 to 0.97 in the potted plants, and from 0.004 to 0.31 at the contaminated marsh site. Stem concentration factor (SCF) values ranged from 0.00004 to 0.03 in the potted plants and 0.0002 to 0.04 at the contaminated marsh. No correlation was found between the RCF value and PAH compound or chemical properties such as logKOW. SCF values were higher for the lighter PAHs in the potted plants, but not in the plants collected from the contaminated marsh. PAH concentrations in the roots of the potted plants are strongly correlated with soil concentrations, but there is less correlation for the roots grown in natural sediments. Additional plants were grown directly in PAH-contaminated water and analyzed for alkylated PAH homologs. No difference was found in leaf PAH concentrations between plants grown in contaminated water and control plants.     

EDTA and urease effects on Hg accumulation by Lepidium sativum

The phytoextraction process was conducted under laboratory conditions with the use of garden cress plants (Lepidium sativum). The experiment was carried out in a model soil, which was characterized before conducting the process. Inorganic forms of mercury (HgCl(2), HgSO(4), Hg(NO(3))(2)) were used for contamination of the soil. The phytoextraction process was conducted after EDTA application to the soil and after urease application. Also the influence of simultaneous addition of ethylenediaminetetraacetic acid (EDTA) and urease into the soil on phytoextraction process was measured. In all variants of phytoextraction process the total mercury concentrations in roots, stems and leaves of garden cress were determined. The result showed that garden cress accumulated mercury from soil. The overall maximum concentration of mercury in its compounds was found in roots of the plant. In all cases, before addition of urease and EDTA, the translocation process and distribution of mercury in the plant tissues were limited. The addition of urease caused an increase of enzyme activity in the soil and at the same time caused an increase of mercury concentration in plant tissues. Application of EDTA increased solubility of mercury and caused an increase of metal accumulation by plants. After simultaneous addition of EDTA and urease into the soil garden cress accumulated about 20% of total mercury concentration in the soil. Most of mercury compounds were accumulated in leaves and stems of the plants (46.0-56.9% of total mercury concentration in the plant tissues).     

Effects of salinity and ozone,individually and in combination,on the growth and ion contents of two chickpea (Cicer arietinum L.) varieties

The aim of this study was to determine the effects of ozone and salinity, singly and in combination, on the growth and ion contents of two chickpea (Cicer arietinum L.) varieties. Chickpea plants were grown in non-saline and saline conditions, with and without a repeated exposure to ozone. Salinity at a concentration of 30 mM NaCl caused a substantial reduction in plant height, number of leaves and the dry weights of the leaves, stems and roots. Biomass allocation to the leaves increased, predominantly at the expense of the roots. Ozone at a concentration of 85 nmol mol(-1) for 6 h per day for 25 days reduced plant height and dry weights but had no effect on leaf number. The results show substantial effects of salinity and ozone on chickpea growth and ion concentrations. When ozonated plants are grown in the presence of salinity, further reductions in growth occur.     

Chemical composition and ecophysiological responses of Empetrum nigrum to aboveground element application

Empetrum nigrum L. (crowberry) is one of the plants surviving near the Cu-Ni smelters in Finland and Russia. According to field observations, the fine roots of E. nigrum are situated below 40 cm depth and the root biomass is reduced in the polluted sites. This could cause a reduced root uptake of macronutrients and trace elements in the field and, therefore, the possible element uptake by aboveground parts of E. nigrum was studied in a greenhouse. Six different treatment solutions containing various heavy metal and macronutrient concentrations were applied to the stems and leaves of E. nigrum and the chemical composition and ecophysiological parameters were measured. Heavy metal concentrations in the leaves and stem bark, and Cu concentrations in the stems, increased with increasing metal concentrations in the spraying solutions. The bark and leaves had higher heavy metal concentrations than the stems of comparable age classes. The macronutrient and Mn concentrations in E. nigrum did not change significantly with increasing element concentrations in the spraying solution. Neither the stem water potential nor the leaf chlorophyll concentrations showed any clear response to element applications. Therefore, the element uptake by aboveground parts of E. nigrum was not confirmed by this study. However, there was a tendency to a decrease in CO2 exchange rate and increase in foliar abscisic acid content in plants treated with the highest element concentrations.