Physiological and Biochemical Mechanisms Preventing Cd Toxicity in the New Hyperaccumulator <Emphasis Type="Italic">Abelmoschus manihot</Emphasis>

Abelmoschus manihot, an ornamental plant, was examined for phytoremediation purposes in accordance with the ability to accumulate cadmium and physiological mechanisms of cadmium tolerance. A net photosynthetic rate (A _N) glasshouse experiment for 60 days was conducted to investigate the influence of different cadmium amounts (0–100 mg kg^?1) on the growth, biomass, photosynthetic performance, reactive oxygen species (ROS) production, antioxidative enzyme activities, Cd uptake and accumulation of A. manihot. Exposure to cadmium enhanced plant growth even at 100 mg kg^?1, without showing symptoms of visible damage. The cadmium concentration of shoots (stems or leaves) and roots was more than the critical value of 100 mg kg^?1 and reached 126.17, 185.26 and 210.24 mg kg^?1, respectively. BCF values of A. manihot plants exceeded the reference value 1.0 for all the Cd treatments, and TF values were greater than 1 at 15–60 mg kg^?1 Cd treatment. The results also showed that cadmium concentrations of 60 mg kg^?1 or less induced a significant enhancement in plant net photosynthetic rate (A _N), stomatal conductance (G _s), transpiration rate (T _r), photosynthetic pigments and F _v/F _m. These parameters were slightly decreased at the higher concentration (100 mg kg^?1). The ROS production (O₂ ^?, H₂O₂) and antioxidative response including SOD, CAT and POD were significantly enhanced by increasing cadmium. These results suggest that A. manihot can be considered as a Cd-hyperaccumulator and the hormetic effects may be taken into consideration in remediation of Cd contamination soil.     

Increased Resistance to Cd(II) in the Primitive Red Algae Cyanidioschyzon merolae

Growth of Cyanidioschyzon merolae was inhibited depending on the cadmium(II) concentration in the culture medium. Although a lower level (0.01 mM) of Cd(II) inhibited growth by a factor of 0.5, higher levels (0.1 and 1 mM) induced lag periods of 10–14 days. Algal cells pretreated with 1 mM Cd(II) for 27 days grew steadily in 1 mM Cd(II) without the lag period, demonstrating that the cells became Cd(II) resistant (CdR). Cells remained resistant after four cycles (7 days per cycle) of washing and re-growing in medium without Cd(II), while intracellular Cd(II) decreased to undetectable levels. These results suggest that the Cd(II)-resistant phenotype is heritable. This phenomena may be attributable to the presence of genetic inhomogeneity in the wild-type cell populations or to mutagenesis caused by Cd(II) stress. Intracellular Cd(II) levels significantly decreased in the CdR phenotype compared to the wild-type cells, indicating that resistant cells may have a defective gene that codes for Cd(II)-uptake protein or the ability to secrete Cd(II).     

Physiological responses of peanut seedlings to exposure to low or high cadmium concentration and the alleviating effect of exogenous nitric oxide to high cadmium concentration stress

Abstract

The physiological responses of peanut seedlings exposed to low (5 µM) or high (200 µM) cadmium (Cd) concentration and the ability of sodium nitroprusside (SNP, a donor of NO) to reverse the harmful effects of Cd on peanut (Arachis hypogaea L.) were studied. Changes in plant growth parameters, chlorophyll content, antioxidant system, nutrient contents and Cd accumulation were investigated. The results showed that SNP and 5 µM Cd improved plant growth and chlorophyll content. Furthermore, antioxidative system was up-regulated, and as a result, the production rate of superoxide radical (O₂^??) was reduced. Moreover, the absorption of nutrient elements was not impacted, and Cd toxicity was not observed. However, 200 µM Cd had negative effects on the above measured parameters and dramatically increased the accumulation of Cd in all the plant organs. In the 200 µM Cd treatment, addition of 250 µM SNP stimulated plant growth and increased chlorophyll content. It also enhanced the regulation of antioxidative system and reduced the production rate of O₂^?? and malondialdehyde (MDA) content. Besides, SNP supply enhanced the absorption of nutrient elements and restrained the absorption and transport of Cd.     

5-Aminolevulinic Acid Ameliorates the Growth, Photosynthetic Gas Exchange Capacity, and Ultrastructural Changes Under Cadmium Stress in Brassica napus L.

Heavy-metal toxicity in soil is one of the major constraints for oilseed rape (Brassica napus L.) production. One of the best ways to overcome this constraint is the use of growth regulators to induce plant tolerance. Response to cadmium (Cd) toxicity in combination with a growth regulator, 5-aminolevulinic acid (ALA), was investigated in oilseed rape grown hydroponically in greenhouse conditions under three levels of Cd (0, 100, and 500 μM) and three levels of foliar application of ALA (0, 12.5, and 25 mg l^?1). Cd decreased plant growth and the chlorophyll concentration in leaves. Foliar application of ALA improved plant growth and increased the chlorophyll concentration in the leaves of Cd-stressed plants. Significant reductions in photosynthetic parameters were observed by the addition of Cd alone. Application of ALA improved the net photosynthetic and gas exchange capacity of plants under Cd stress. ALA also reduced the Cd content in shoots and roots, which was elevated by high concentrations of Cd. The microscopic studies of leaf mesophyll cells under different Cd and ALA concentrations showed that foliar application of ALA significantly ameliorated the Cd effect and improved the structure of leaf mesophyll cells. However, the higher Cd concentration (500 μM) could totally damage leaf structure, and at this level the nucleus and intercellular spaces were not established as well; the cell membrane and cell wall were fused to each other. Chloroplasts were totally damaged and contained starch grains. However, foliar application of ALA improved cell structure under Cd stress and the visible cell structure had a nucleus, cell wall, and cell membrane. These results suggest that under 15-day Cd-induced stress, application of ALA helped improve plant growth, chlorophyll content, photosynthetic gas exchange capacity, and ultrastructural changes in leaf mesophyll cells of the rape plant.     

Immunological and cytotoxicological responses of the Asian clam,Corbicula fluminea (M.), experimentally exposed to cadmium

Abstract

Bivalve molluscs, as filter-feeding organisms, are known to accumulate metals that can produce deleterious effects on organisms. The phagocytic activity of haemocytes and lysosomal alterations in the digestive gland cells were measured in the freshwater Asian clam exposed to cadmium, in order to assess the possible use of immunocompetence and lysosomal responses as biomarkers of freshwater quality. Clams were exposed in the laboratory to nominal concentrations of 3, 10, 21.4, 46.5 and 100 µg l^?1 of cadmium and sampled after 7, 15 and 30 days of exposure. The results show a decrease of phagocytic activity after only 7 days of exposure to 10 µg l^?1 of cadmium. This response was also observed as the exposure time was increased. Lysosomes in the digestive cells increased in size and number after 7 days of exposure as cadmium concentration increased. After 30 days of exposure, a decrease in size and number indicated a change in the response to the metal from concentrations of 46.5 µg l^?1 of cadmium. A dose and time response both in phagocytic activity of haemocytes and lysosomal structure demonstrated a possible use of these biomarkers in freshwater biomonitoring.     

Responses of Lemna trisulca L. (Duckweed) exposed to low doses of cadmium: thiols, metal binding complexes, and photosynthetic pigments as sensitive biomarkers of ecotoxicity

Lemna species are reported to accumulate a variety of metals from contaminated/polluted sites. Cadmium is a nonessential element for plant metabolism. In this work, we aimed to investigate physiological responses to low doses of cadmium (up to 100 μM). From exposure to the lowest Cd concentration (1 µM) to the highest (100 µM), photosynthetic pigments (Chl a, b, carotenoids) and the ratios of Chl a/b, Chl (a + b)/carotenoids decreased as a function of the Cd dose. The content of soluble proteins decreased in a dose-dependent manner, while total soluble thiols drastically increased. In Cd-treated fronds, the dose-dependent accumulation of a polypeptide with an apparent molecular weight of 24 kDa, as well as the appearance of two smaller polypeptides with molecular weights <6.5 kDa, was observed in sodium dodecyl sulfate polyacrylamide gel electrophoresis. Our results show that in Lemna trisulca, different adaptative mechanisms may be involved in counterbalancing low and high doses of a particular toxicant (cadmium). This feature makes this plant potentially useful material in biomonitoring and phytotoxicity testing.     

Testicular Histomorphometry and Ultrastructure of Rats Treated with Cadmium and Ginkgo biloba

The aim of this study is to investigate the association of a single low dose of Cd and daily doses of Ginkgo biloba extract (GbE) on the testis and accessory glands of rats. The animals were treated with a single dose of 3 µmol/kg body weight of cadmium chloride (CdCl₂) and/or 100 mg/kg body weight of GbE. The plasma testosterone levels; corporal, testicular, and accessory glands weight; gonadosomatic index, volumetric proportion; and absolute volume of testicular components did not change after the treatments. CdCl₂ caused significant reduction in Leydig cells volume and altered Leydig cell morphology, as well as vacuolated Sertoli cells cytoplasm, irregular chromatin condensation of late spermatids, and modified acrosome formation. However, animals that received GbE did not show these alterations. The reversal of Cd-induced alterations by the extract is a strong indication that G. biloba is helpful in diminishing the effect of Cd toxicity.     

Reactive oxygen species formation and cell death in catalase-deficient tobacco leaf disks exposed to cadmium

The physiological responses of tobacco (Nicotiana tabacum L.) to oxidative stress induced by cadmium were examined with respect to reactive oxygen species (ROS) formation, antioxidant enzymes activities, and cell death appearance in wild-type SR1 and catalase-deficient CAT1AS plants. Leaf disks treated with 100 or 500 µM CdCl₂ increased Evans blue staining and leakage of electrolytes in SR1 or CAT1AS plants, more pronouncedly in the transgenic cultivar, but without evidence of lipid peroxidation in any of the cultivars compared to controls. Cadmium significantly reduced the NADPH oxidase-dependent O ₂ ^? formation in a dose dependent manner in SR1 very strongly at 500 µM (to 5% of the activity in the nontreated SR1 leaf disks). In CAT1AS, the NADPH oxidase activity was constitutively reduced at 50% with respect to that of SR1, but the magnitude of the decay was less prominent in this cultivar, reaching an average of 64% of the C at 21 h, for both Cd concentrations. Hydrogen peroxide formation was only slightly increased in SR1 or CAT1AS leaf disks at 21 h of exposure compared to the respective controls. Cd increased superoxide dismutase activity more than six times at 21 h in CAT1AS, but not in SR1 and reduced catalase activity by 59% at 21 h of treatment only in SR1 plants. Despite that catalase expression was constitutively lower in CATAS1 compared to SR1 nontreated leaf disks, 500 µM CdCl₂ almost doubled it only in CAT1AS at 21 h. The mechanisms underlying Cd-induced cell death were possibly not related exclusively to ROS formation or detoxification in tobacco SR1 or CAT1AS plants.     

Cadmium effects and accumulation in cultures ofProrocentrum micans (dinophyta)

Effects and accumulation of cadmium were studied in unialgal 10-1 batch-culture experiments with the dinoflagellateProrocentrum micans Ehrenberg. Tests were made using sterile filtered North Sea water enriched with nitrate and phosphate only in order to avoid disturbances by complex formation. Cadmium was added to the cultures in amounts of 100 to 0.13μg l⁻¹. In one series it was added at the start of the experiments and in a second one after a growth period of 1 week. Addition of only 1.2μg Cd l⁻¹ reduces multiplication rates and maximum cell densities of the algae. Not until 0.4μg Cd⁻¹ does growth correspond to that of the controls. Cadmium concentrations were measured, after filtration, in the culture medium and in the biomass by means of flameless AAS. The cadmium content in algae increased from 2.7μg g⁻¹ (dry weight) in controls to 500μg g⁻¹ (dry weight) in media containing 100μg Cd l⁻¹. Uptake occurred rapidly during the first few days of the experiments, slowed down somewhat during exponential growth stage, and increased during decay of the cultures. Cadmium content of culture media remained nearly constant (Series 1) or decreased only slowly during experimental time (Series 2). The highest concentration factor was measured in the controls. It decreased with increasing metal concentration in the medium and increased with experimental time. Structural modifications of the cells were visible after Lugol fixation only, indicating brittleness of the cell walls.P. micans has shown to be extremely sensitive to cadmium and to accumulate this metal.     

Melatonin affects the growth and cadmium accumulation of Malachium aquaticum and Galinsoga parviflora

Phytoremediation technology has become one of the main techniques for remediating soils polluted by heavy metals because it does not damage the environment, but heavy metal-tolerant plants have the disadvantages of low biomass and slow growth. A pot experiment was conducted to study the effects of melatonin (Mel) on growth and cadmium (Cd) accumulation in the Cd accumulator Malachium aquaticum and hyperaccumulator Galinsoga parviflora by spraying different concentrations of Mel on them. The results showed that shoot biomass, photosynthetic pigment content and antioxidant enzyme activity were increased in both species after Mel was sprayed on their leaves. Mel reduced the Cd content in shoots of M. aquaticum and increased it in those of G. parviflora. In general, Cd accumulation was greatest in M. aquaticum when Mel was 200 μmol L^?1 (120.71 μg plant^?1, increased by 15.97% than control) and in G. parviflora when Mel was 100 μmol L^?1 (132.40 μg plant^?1, increased by 68.30% than control). Our results suggest it is feasible to improve the remediation efficiency of lightly Cd-contaminated soil by spraying G. parviflora with100 μmol L^?1 Mel.     

Cadmium and zinc uptake,growth, and primary production inCoscinodiscus granii cultures containing low levels of cells and dissolved organic carbon

Coscinodiscus granii Gough was cultivated at low cell densities in aged Atlantic sea water containing very little dissolved organic carbon; the water was enriched with low levels of nutrients but no chelators were added. Cadmium additions provided final concentrations of 0.1 to 26.5 μg Cd l⁻¹, zinc being kept constant at a level of 38 μg Zn l⁻¹. Carrier-free¹⁰⁹Cd and⁶⁵Zn were used as tracers for the two metals. Growth in terms of cell numbers and primary productivity capacity, using the¹⁴C uptake rate, was followed during the exponential growth phase for the first 5 days of the experiment and then for a further 3 days during the stationary phase. On each day, the metal contents of the cells were determined. Cadmium concentrations of 20 μg Cd l⁻¹ and more resulted in growth inhibition whereas 17.5 μg Cd l⁻¹ reduced the growth only slightly. The physiological state of the cells influenced the heavy-metal uptake per cell at sublethal Cd levels. Dead cells had a higher heavy-metal concentration than living cells. Microscopical observations revealed that cells just about to divide were less sensitive to a given toxic heavy-metal concentration than cells which had recently divided. This might have been due to different surface/volume ratios.     

Effects of Cadmium Stress on Leaf Chlorophyll Fluorescence and Photosynthesis of Elsholtzia argyi—A Cadmium Accumulating Plant

A hydroponic experiment was conducted to investigate the effects of cadmium (Cd) on chlorophyll fluorescence and photosynthetic parameters on a Cd accumulating plant of Elsholtzia argyi. Four weeks-seedlings of E. argyi were treated with 0 (CK) 5, 10, 15, 20, 25, 30, 40, 50 and 100 μmol L^?1 Cd for 21days. Fv/Fo, Fv/Fm, qP, ΦPSП, ETR and Fv′/Fm′ were significantly increased under low Cd (5–15 μmol L^?1 for Fv/Fo, Fv/Fm and qP, 5–10 μmol L^?1 for ΦPSП, ETR and Fv′/Fm′) stress, and these parameters were similar to control under Cd ≤ 50μmol L^?1. All above parameters were significantly decreased at 100 μmol L^?1 Cd. Compared with control, Pn was significantly (P < 0.05) increased under 5–30 μmol L^?1 Cd. However, 50 and 100 μmol L^?1 Cd significantly (P < 0.05) reduced it. Gs and Tr were substantially decreased at 50–100 and 40–100 μmol L^?1 Cd, respectively. Ci was significantly increased at 50 and 100 μmol L^?1 Cd. High Cd-induced decrease of Pn is not only connected to stomatal limitation but also to the inhibition of Fv/Fo, Fv/Fm, ΦPSП, qP, ETR and increase of NPQ. Maintain chlorophyll fluorescence and photosynthesis parameters under its Cd tolerance threshold were one of tolerance mechanisms in E. argyi.     

Environmental Cadmium Exposure Impacts Physiological Responses in Manila clams

The physiological responses of marine bivalves to chronic cadmium (Cd) exposure at sub-lethal concentrations have been well documented. As of now, few studies have examined the effect of Cd exposure and subsequent recovery period at environmentally realistic concentrations. In this study, environmentally, Cd exposures were performed to assess the physiological responses of the Manila clam Ruditapes philippinarum. The clams were exposed to waterborne Cd at two environmentally realistic concentrations (4 and 40 μg L^?1) for 35 days and then allowed to recover for another 35 days. The accumulation and elimination of Cd in R. philippinarum were tissue-specific and dose- and time-dependent. Cd accumulation increased sharply in the digestive gland, and Cd elimination was rapid in the gill. Major physiological responses, including clearance rate, absorption efficiency, respiration rate, excretion rate, oxygen to nitrogen ratio, and scope for growth, were significantly affected by Cd exposure. Yet, the clams exposed to 4-μg L^?1 Cd were able to quickly recover their normal physiological processes and clearly exhibited catch-up growth once they were transferred to clean seawater. Hence, R. philippinarum can exhibit good physiological plasticity when confronted with moderately environmental Cd exposure. All physiological responses measured exhibited a highly significant and generally predictable correlation with tissue Cd concentration, which in turn, reflected environmentally realistic exposure conditions. Our results further confirm that the measurement of physiological responses is a sensitive method for assessing stress at environmentally realistic metal concentrations.     

Cadmium Removal Potential from Contaminated Soil by Tagetes erecta L. and Panicum maximum: Influence of Soil pH

The potential for cadmium (Cd) removal from contaminated soil by two species—marigold (Tagetes erecta L.) and Guinea grass (Panicum maximum)—was investigated in pot culture experiments in a greenhouse in triplicate. The concentration of Cd was varied from 50 to 200 mg kg^?1 and the pH was varied from 5.0 to 7.5 to investigate the effect of pH on Cd uptake. The results showed that total biomass of Guinea grass was around nine and seven times higher than that of marigold for Cd treatments of 50 and 100 mg kg^?1 at pH 5.0, respectively. Total cadmium uptake at Cd treatments of 50 and 100 mg kg^?1 at pH 5.0 by Guinea grass was 19.28 ± 3.14 and 36.06 ± 4.28 mg kg^?1, respectively, and for marigold was 15.66 ± 4.17 and 20.38 ± 3.24 mg kg^?1, respectively. The total Cd uptake by Guinea grass was 1.23 and 1.77 higher than that of marigold at Cd treatments of 50 and 100 mg kg^?1, respectively, at pH 5.0 due to higher biomass. The maximum Cd uptake by marigold and Guinea grass occurred at pH 5.0 at Cd treatment of 100 mg kg^?1. The results clearly show that the two species behave very differently for Cd uptake. Guinea grass is easy to grow, drought tolerant and, due to its higher biomass, it can be used for remediation of Cd-contaminated soil.     

Matricaria chamomilla is not a hyperaccumulator, but tolerant to cadmium stress

The influence of low (3 μM) and high (60 and 120 μM) cadmium (Cd) concentrations were studied on selected aspects of metabolism in 4-week-old chamomile (Matricaria chamomilla L.) plants. After 10 days’ exposure, dry mass accumulation and nitrogen content were not significantly altered under any of the levels of Cd. However, there was a significant decline in chlorophyll and water content in the leaves. Among coumarin-related compounds, herniarin was not affected by Cd, while its precursors (Z)- and (E)-2-β-d-glucopyranosyloxy-4-methoxycinnamic acids (GMCAs) increased significantly at all the levels of Cd tested. Cd did not have any effect on umbelliferone, a stress metabolite of chamomile. Lipid peroxidation was also not affected by even 120 μM Cd. Cd accumulation was approximately seven- (60 μM Cd treatment) to eleven- (120 μM Cd treatment) fold higher in the roots than that in the leaves. At high concentrations, it stimulated potassium leakage from the roots, while at the lowest concentration it could stimulate potassium uptake. The results supported the hypothesis that metabolism was altered only slightly under high Cd stress, indicating that chamomile is tolerant to this metal. Preferential Cd accumulation in the roots indicated that chamomile could not be classified as a hyperaccumulator and, therefore, it is unsuitable for phytoremediation.     

Cadmium induced changes in subcellular glutathione contents within glandular trichomes of Cucurbita pepo L.

Plants cope with cadmium (Cd) stress by complexation with phytochelatins (Pc), metallothioneins and glutathione and sequestration within vacuoles. Especially glutathione was found to play a major role in Cd detoxification as Cd shows a high binding affinity towards thiols and as glutathione is a precursor for Pc synthesis. In the present study, we have used an immunohistochemical approach combined with computer-supported transmission electron microscopy in order to measure changes in the subcellular distribution of glutathione during Cd-stress in mesophyll cells and cells of different glandular trichomes (long and short stalked) of Cucurbita pepo L. subsp. pepo var. styriaca Greb. Even though no ultrastructural alterations were observed in leaf and glandular trichome cells after the treatment of plants with 50 µM cadmium chloride (CdCl₂) for 48 h, all cells showed a large decrease in glutathione contents. The strongest decrease was found in nuclei and the cytosol (up to 76%) in glandular trichomes which are considered as a major side of Cd accumulation in leaves. The ratio of glutathione between the cytosol and nuclei and the other cell compartments was strongly decreased only in glandular trichomes (more than 50%) indicating that glutathione in these two cell compartments is especially important for the detoxification of Cd in glandular trichomes. Additionally, these data indicate that large amounts of Cd are withdrawn from nuclei during Cd exposure. The present study gives a detailed insight into the compartment-specific importance of glutathione during Cd exposure in mesophyll cells and glandular trichomes of C. pepo L. plants.     

In vitro biochemical evaluation of cadmium tolerance mechanism in callus and seedlings of Brassica juncea

In vitro grown callus and seedlings of Brassica juncea were treated with equimolar concentrations of cadmium and compared for their respective tolerance to cadmium. Calli cultures were grown on Murashige and Skoog medium supplemented with α 6-benzyl aminopurine (200 µg L^?1, naphthalene acetic acid 200 µg L^?1) and 2,4-dichloro-phenoxy acetic acid (65 µg L^?1) while the seedlings grown on Hoagland's nutrient solution have been carried out. Cellular homeostasis and detoxification to cadmium in B. juncea were studied by analyzing the growth in terms of fresh weight and dry weight, lipid peroxidation, proline accumulation, and antioxidative enzymes (superoxide dismutase (SOD), ascorbate peroxidase (APX), and catalase (CAT)). At 200 µM cadmium, callus and seedlings showed 73.61% and 74.76% reduction in tolerance, respectively. A significant increase in malondialdehyde (MDA) content was found in both calli and seedlings; however, the amount of MDA content was more in seedlings. Proline content increased on lower concentration of cadmium (up to 50 µM), and it further decreased (up to 200 µM). But the accumulation of proline was higher in callus cultures. The overall activity of antioxidative enzymes (SOD, CAT, and APX) was found to be higher in callus in comparison to seedlings of B. juncea. Callus and seedlings showed a significant (P?≤?0.5) increase in SOD activity in a concentration-dependent manner up to 50 µM cadmium concentration but decreased further. APX activity increased significantly at low cadmium levels but CAT activity decreased significantly throughout on increasing cadmium concentrations from 5 to 200 µM, respectively. Hence, it was observed that callus of B. juncea was more tolerant in comparison to seedlings exposed to equimolar concentrations of cadmium. Thus, from the present studies, it is concluded that calli were more tolerant toward cadmium-induced oxidative stress. Hence, it is suitable material for the study of cadmium tolerance mechanisms and for the manipulations within them for better understanding of cadmium detoxification strategies in B. juncea.     

Bio-removal of cadmium by growing deep-sea bacterium Pseudoalteromonas sp. SCSE709-6

Effective bio-removal of heavy metals is important for water treatment. Although a number of microorganism species demonstrated the ability of living cells to remove cadmium, most of them were tested at fixed concentration of metals, salinity, and temperature. This paper reported a research on the screening and performance of a newly developed deep-sea bacterium, Pseudoalteromonas sp. SCSE709-6, for Cd(II) removal by growing cells under a range of experimental conditions: 0–50 mg/L of Cd(II), 15–30 °C of incubation temperatures, 6.5–8.0 of initial pH, and 1.5–5.0 % of salinity. Study results revealed that Pseudoalteromonas sp. SCSE709-6 could remove more than 96 % of Cd(II) on growth. The Cd(II) bioremoval was in correlation but not in accordance with biomass. As cadmium concentrations increased, the Cd(II) removal by cell adsorption played an increasingly important role compared with that of intracellular accumulation. For the removal mechanism, Fourier transform infrared spectroscopy revealed that carboxyl, amido and hydroxyl of saccharides, and proteins in the extracellular polymeric substances are the most active groups for Cd(II) absorption. The bacterium reported in this study offers a new microbe strain for Cd(II) bioremediation.     

Enhancement of phenylpropanoid enzymes and lignin in Phalaenopsis orchid and their influence on plant acclimatisation at different levels of photosynthetic photon flux

Effects of three levels of photosynthetic photon flux (PPF: 60, 160 and 300 μmol m⁻²s⁻¹) were investigated in one-month-old Phalaenopsis plantlets acclimatised ex vitro. Optimal growth, chlorophyll and carotenoid concentations, and a high carotenoid:chlorophyll a ratio were obtained at 160 μmol m⁻²s⁻¹, while net CO₂ assimilation (A), stomatal conductance (g), transpiration rate (E) and leaf temperature peaked at 300 μmol m⁻²s⁻¹, indicating the ability of the plants to grow ex vitro. Adverse effects of the highest PPF were reflected in loss of chlorophyll, biomass, non-protein thiol and cysteine, but increased proline. After acclimatisation, glucose-6-phosphate dehydrogenase, shikimate dehydrogenase, phenylalanine ammonia-lyase (PAL) and cinnamyl alcohol dehydrogenase (CAD) increased, as did lignin. Peroxidases (POD), which play an important role in lignin synthesis, were induced in acclimatised plants. Polyphenol oxidase (PPO) and β-glucosidase (β-GS) activities increased to a maximum in acclimatised plants at 300 μmol m⁻²s⁻¹. A positive correlation between PAL, CAD activity and lignin concentration was observed, especially at 160 and 300 μmol m⁻²s⁻¹. The study concludes that enhancement of lignin biosynthesis probably not only adds rigidity to plant cell walls but also induces defence against radiation stress. A PPF of 160 μmol m⁻²s⁻¹was suitable for acclimatisation when plants were transferred from in vitro conditions.     

Effects of biopellets composed of microalgae and fungi on cadmium present at environmentally relevant levels in water

Removal of cadmium (Cd) present at low levels (1 µg L^?1) in water was investigated using three different microorganism treatments: the microalga Chlorella vulgaris, the fungus Aspergillus niger, and for the first time biopellets composed of C. vulgaris and A. niger. After 5 days, all microorganism treatments resulted in significantly lower Cd concentrations compared with the control. Biopellets treatment resulted in significantly lower pH values than other microorganism treatments, indicating that the biopellets should be investigated further for their water treatment capacity, since pH is a factor affecting the fate of numerous pollutants in water. Use of biopellets as a method to simplify harvesting of microalgae from water is also suggested.