Jiřina Száková

Czechoslovakian biological certified reference materials and their use in the analytical quality assurance system in a trace element laboratory

SummaryThe Trace Element Laboratory of the University of Agriculture in Prague (CSFR) participated successfully in interlaboratory experiments on the determination of the contents of trace elements in seven Czechoslovakian reference materials. Czechoslovakian certified reference material CRM 12-02-01 (Bovine liver) was used for the development of a new analytical method using the Dry Mineralizer Apion as well as in analytical quality assessment of data in the determination of Cd, Pb, and Hg contents in routinely analyzed animal tissues.

Evaluation of DNA damage and mutagenicity induced by lead in tobacco plants

Tobacco (Nicotiana tabacum L. var. xanthi) seedlings were treated with aqueous solutions of lead nitrate (Pb2+) at concentrations ranging from 0.4 mM to 2.4 mM for 24 h and from 25 microM to 200 microM for 7 days. The DNA damage measured by the comet assay was high in the root nuclei, but in the leaf nuclei a slight but significant increase in DNA damage could be demonstrated only after a 7-day treatment with 200 microM Pb2+. In tobacco plants growing for 6 weeks in soil polluted with Pb2+ severe toxic effects, expressed by the decrease in leaf area, and a slight but significant increase in DNA damage were observed. The tobacco plants with increased levels of DNA damage were severely injured and showed stunted growth, distorted leaves and brown root tips. The frequency of somatic mutations in tobacco plants growing in the Pb2+-polluted soil did not significantly increase. Analytical studies by inductively coupled plasma optical emission spectrometry demonstrate that after a 24-h treatment of tobacco with 2.4 mM Pb2+, the accumulation of the heavy metal is 40-fold higher in the roots than in the above-ground biomass. Low Pb2+ accumulation in the above-ground parts may explain the lower levels or the absence of Pb2+-induced DNA damage in leaves.

The Rengen Grassland Experiment: relationship between soil and biomass chemical properties, amount of elements applied, and their uptake

The Rengen Grassland Experiment (RGE) was established in the Eifel Mountains (Germany) on a low productive Nardetum in 1941. Since then, the following fertilizer treatments have been applied with a late two-cut system: unfertilized control, Ca, CaN, CaNP, CaNPKCl and CaNPK2SO4. We aimed to understand how concentrations of macro (N, P, K, Ca and Mg), micro (Cu, Fe, Mn and Zn) and trace (As, Cd, Cr, Ni and Pb) elements in the plant biomass were affected by long-term fertilizer application, soil chemical properties and biomass production. In 2008, biomass samples from the first cut (early July) and the second cut (mid-October) were collected and analyzed. The simultaneous application of N, P and K decreased nitrogen concentration in the aboveground biomass, but substantially increased biomass production. Late cutting management decreased forage quality in highly productive more than in low productive plant communities. The concentrations of P and K in the plant biomass were positively related to P and K application and, therefore, to plant available P and K concentrations in the soil. The concentrations of some micro (Fe, Mn and Zn) and trace (As, Cd, Cr, Ni and Pb) elements in the plant biomass were negatively correlated with the amount of elements supplied by fertilizers and biomass production, probably because of the dilution effect. Long-term fertilizer application resulted in the accumulation of macro (P, Ca and Mg), micro (Fe and Mn) and trace (As and Cr) elements in the soil, but in many cases this accumulation was not connected with an increase in the concentrations of these elements in the plant biomass. Nutritional status, as indicated by the biomass N:P ratio, was consistent with N or P limitation as indicated by the nitrogen and phosphorus nutrition indices. Furthermore, additional K (co-)limitation was indicated by the N:K and K:P ratios in the biomass from the NP treatment. The results from the RGE indicate that there is no simple positive relationship between the applied elements and their concentrations in the plant biomass.

Potential and drawbacks of EDDS-enhanced phytoextraction of copper from contaminated soils.

Incubation and pot experiments using poplar (Populus nigra L. cv. Wolterson) were performed in order to evaluate the questionable efficiency of EDDS-enhanced phytoextraction of Cu from contaminated soils. Despite the promising conditions of the experiment (low contamination of soils with a single metal with a high affinity for EDDS, metal tolerant poplar species capable of producing high biomass yields, root colonization by mycorrhizal fungi), the phytoextraction efficiency was not sufficient. The EDDS concentrations used in this study (3 and 6 mmol kg(-1)) enhanced the mobility (up to a 100-fold increase) and plant uptake of Cu (up to a 65-fold increase). However, despite EDDS degradation and the competition of Fe and Al for the chelant, Cu leaching cannot be omitted during the process. Due to the low efficiency, further research should be focused on other environment-friendly methods of soil remediation.

The effect of arsenic contamination on amino acids metabolism in Spinacia oleracea L.

Changes of amino acid concentrations (proline, glutamate, asparagine, aspartate, alanine) and glutamate kinase activity (GKA) in plants under arsenic chronic stress reported here reveal their role in plant arsenic stress adaptation. Results of the pot experiment confirmed the toxic effect of arsenic at tested levels (As1=25 mg As kg(-1) soil, As2=50 mg As kg(-1) soil, As3=75 mg As kg(-1) soil) for spinach. Growing available arsenic contents in soil were associated with the strong inhibition of above-ground biomass and with the enhancement of As plant content. The changes of glutamate, asparagine, aspartate and proline levels in the plants showed strong linear dependences on arsenic concentration in plants (R2=0.60-0.90). Compared to the untreated control, concentrations of free proline and aspartate of As3 treatment were enhanced up to 381% and 162%, respectively. The significant changes of glutamate were observed on As2 and As3 treatments (increased level up to 188, i.e. 617%). Arsenic in plants was shown to be an inhibitor of glutamase kinase activity (R2=0.91). Inhibition of GKA resulted in an increase in the content of glutamate that is used in synthesis of phytochelatins in plant cells. Concentration of alanine did not have a confirmed linear dependence on arsenic concentration in plant (R2=0.05). The changes of its concentrations could be affected by changes of pH in plant cell or induction of alanine aminotransferase by hypoxia.

Retention of copper originating from different fungicides in contrasting soil types

This work described the retention of Cu from two different commonly used pesticides, the Bordeaux mixture (CuSO(4)+Ca(OH)(2)) and Cu-oxychloride (3Cu(OH)(2).CuCl(2)), and from Cu(NO(3))(2) in contrasting soil types (Leptosol, Chernozem, Cambisol). Thermodynamic modeling showed that Cu speciation was similar in all fungicide solutions. However, the retention of Cu differed with the fungicide used (maximum retention from the Bordeaux mixture) which indicates that different retention processes occurred in the studied soils. The suggested mechanisms include: specific and non-specific adsorption (especially on soil organic matter), precipitation of newly formed phases, such as CuO, Cu(OH)(2), Cu(2)(OH)(3)NO(3), CuCO(3)/Cu(2)(OH)(2)CO(3) and in the case of the Bordeaux mixture, precipitation of various Cu-hydroxysulfates. These phases were identified by the speciation model. The retention of fungicide-derived Cu in the studied soil types followed well the Freundlich isotherm and was directly controlled by the chemical form of Cu. This fact should be taken into account for both environmental and practical applications.

The use of water lettuce (Pistia stratiotes L.) for rhizofiltration of a highly polluted solution by cadmium and lead.

The effectiveness of heavy metal uptake from contaminated nutrient solution by four aquatic macrophytes (Pistia stratiotes L., Salvinia auriculata Aubl., Salvinia minima Baker, and Azolla filiculoides Lam) was estimated in this study. The influence of cadmium (3.5 mg L−1 and 10.5 mg L−1) and lead (25 mg L−1 and 125 mg L−1) on the stress symptoms was observed through the determination of chlorophyll content and transpiration rate over 14 days of the experiment. The results of the present study showed extreme reductions in Cd and Pb concentrations in solution during the first 4 days. The accumulation of Pb in plant tissues was the highest during the first 4 days and was more than 10 times higher in the roots (42,862 mg kg−1) than in the leaves (3,867 mg kg−1). The accumulation of Cd slowly increased and was the highest at the end of the experiment. Concentrations in roots (3,923 mg kg−1) were roughly 6 times higher than in the leaves (624 mg kg−1). Results showed significant decrease in the transpiration rate at Pb treatment and a significant increase at Cd treatment during 48 hours of exposition.

Mobility of arsenic and its compounds in soil and soil solution: the effect of soil pretreatment and extraction methods.

The effect of soil extraction procedures and/or sample pretreatment (drying, freezing of the soil sample) on the extractability of arsenic and its compounds was tested. In the first part, five extraction procedures were compared with following order of extractable arsenic portions: 2M HNO(3)>>0.43 M CH(3)COOH>or=0.05 M EDTA>or=Mehlich III (0.2M CH(3)COOH+0.25 M NH(4)NO(3)+0.013 M HNO(3)+0.015 M NH(4)F+0.001 M EDTA) extraction>>water). Additionally, two methods of soil solution sampling were compared, centrifugation of saturated soil and the use of suction cups. The results showed that different sample pretreatments including soil solution sampling could lead to different absolute values of mobile arsenic content in soils. However, the interpretation of the data can lead to similar conclusions as apparent from the comparison of the soil solution sampling methods (r=0.79). For determination of arsenic compounds mild extraction procedures (0.05 M (NH(4))(2)SO(4), 0.01 M CaCl(2), and water) and soil solution sampling using suction cups were compared. Regarding the real soil conditions the extraction of fresh samples and/or in situ collection of soil solution are preferred among the sample pretreatments and/or soil extraction procedures. However, chemical stabilization of the solutions should be allowed and included in the analytical procedures for determination of individual arsenic compounds.

Interactions of EDDS with Fe- and Al-(hydr)oxides.

The efficiency of EDDS ([S,S]-ethylenediaminedisuccinate) in metal (phyto) extraction has been discussed in many recent papers. This study demonstrated that the presence of Fe- and Al-(hydr)oxides in soils influences the speciation of EDDS and thus can decrease the extraction of the targeted metallic contaminants (e.g., Pb, Cu, Zn). Above all, amorphous and poorly crystalline oxides (e.g., ferrihydrite) seem to significantly control dissolved Fe and Al concentrations in soils in the presence of metal-EDDS complexes and especially uncomplexed EDDS. Metals released from these minerals compete for the chelating agent and the extraction efficiency of the targeted metals is lowered. The formation of stable Cu-EDDS complexes, which are preferentially formed in soils with high Cu concentrations, results into a lower dissolution of ferrihydrite and goethite compared to free EDDS and Al-EDDS. Information about the contents of amorphous and poorly crystalline oxides in the treated soils would thus be beneficial for choosing efficient EDDS dosages.

Trace elements present in airborne particulate matter—Stressors of plant metabolism

Changes of amino acid concentrations (glutamic acid, glutamine, asparagine, aspartate, proline, tryptophan, alanine, glycine, valine and serine), gas-exchange parameters (net photosynthetic rate, transpiration rate, stomatal conductance and intercellular CO(2) concentration) and nitrate levels in Lactuca serriola L. under airborne particulate matter (PM) contamination reported here reveal their role in plant chronic stress adaptation. Results of the pot experiment confirmed the toxic effect of trace elements present in PM for lettuce. PM applied to soil or on the lettuce leaves were associated with the strong inhibition of above-ground biomass and with the enhancement of plant trace element contents. The significant changes of amino acid levels and leaf gas-exchange parameters of the plants showed strong linear dependences on PM contamination (R(2)=0.60-0.99). PM application on leaves intensified toxic effect of trace elements (As, Pb, Cr and Cd) originating from PM by shading of the leaf surface. The plant accumulation of nitrate nitrogen after PM contamination confirmed to block nitrate assimilation.