Evaluation of genotoxic variations in plant model systems in a case of metal stressors.

This study was designed to assess the effects of long term, high metal exposition (cadmium, lead, copper, nickel and zinc) on DNA damage in four plant model systems [Taraxacum officinale (Asteraceae), Matricaria recutita L. (Asteraceae), Robinia pseudoacacia L. (Fabaceae), and Urtica dioica (Urticaceae)]. DNA stability was investigated by a Random Amplified Polymorphic DNA (RAPD) technique. Agarose-gel electrophoresis revealed total of 37 bands with different molecular weights ranging from 1250 to 5000 bp. It generated distinctive polymorphism value of 72.97% (27 bands) total in four plant species investigated. The dendrogram constructed using NTSYSpc programme showed that there is grouping in separate clusters of the same plant model collected from two different areas (metal-exposed and control samples). The study concluded that the long term metal-exposing periods had genotoxic stress on macromolecules of plant model systems investigated and biomarkers used should be augmented for reliable estimates of genotoxicity after exposure of plants to metal stressors.

Influence of heavy metal stress on antioxidant status and DNA damage in Urtica dioica.

Heavy metals have the potential to interact and induce several stress responses in the plants; thus, effects of heavy metal stress on DNA damages and total antioxidants level in Urtica dioica leaves and stems were investigated. The samples are sampled from areas with different metal exposition. Metal content was analyzed by Inductively Coupled Plasma-Atomic Emission Spectrometer (ICP-AES), for total antioxidants level assessment the Ferric-Reducing Antioxidant Power (FRAP) assay was used, and genomic DNA isolation from frozen plant samples was performed to obtain DNA fingerprints of investigated plant. It was found that heavy metal contents in stems generally changed synchronously with those in leaves of the plant, and extraneous metals led to imbalance of mineral nutrient elements. DNA damages were investigated by Random Amplified Polymorphic DNA (RAPD) technique, and the results demonstrated that the samples exposed to metals yielded a large number of new fragments (total 12) in comparison with the control sample. This study showed that DNA stability is highly affected by metal pollution which was identified by RAPD markers. Results suggested that heavy metal stress influences antioxidant status and also induces DNA damages in U. dioica which may help to understand the mechanisms of metals genotoxicity.

Mineral nutrient imbalance, total antioxidants level and DNA damage in common bean (Phaseolus vulgaris L.) exposed to heavy metals.

The present study aimed to analyze the biological effects induced by bioaccumulation of metals in common bean (Phaseolus vulgaris L.). Effects of mineral nutrient imbalance, total antioxidants level and DNA damage induced by accumulation of heavy metals, were investigated in bean seedlings treated with two selected metal concentrations for 7 days. Metal content is analyzed by inductively coupled plasma - atomic emission spectrometer (ICP-AES), for total antioxidants level assessment the Ferric-Reducing Antioxidant Power (FRAP) assay is used and Random Amplified Polymorphic DNA (RAPD) method was applied for investigation of DNA damages. The increasing metal concentration in the treatment medium changed synchronously metal content in samples, and decreased total antioxidant activity in all samples with exception only for samples treated with Ni and Cd. The obtained "DNA fingerprints" demonstrated that the increasing metal concentrations induced changes in RAPD profiles (disappearance and/or appearance of bands in comparison with untreated control samples). The highest number of missing bands was observed in samples treated with zinc (total 4 bands) and nickel (total 4 bands) at both concentrations. These results suggested that mineral nutrient imbalance is involved in changes of antioxidant levels and DNA damages of the seedlings, which may help to understand the mechanism of metal toxicity in plants.

Assessment of the genotoxicity of heavy metals in Phaseolus vulgaris L. as a model plant system by Random Amplified Polymorphic DNA (RAPD) analysis.

Impact assessments of environmental pollutants are important in eco-genotoxicology. A random amplified polymorphic DNA (RAPD) technique was used to detect genotoxicity-induced DNA damage in Phaseolus vulgaris L. from heavy metals at two different concentrations. The results from six 10-base pair (bp) random RAPD primers with 60-70% GC content used, showed a total of 295 RAPD fragments of 700-4000 bp in molecular size in the seedlings of untreated and treated samples, of which only 163 fragments were polymorphic. Polymorphisms became evident as the disappearance and/or appearance of DNA fragments in treated samples compared to the control. A dendrogram constructed using the Numerical Taxonomy and Multivariate Analysis System (NTSYSps), showed that the control group merged with groups treated with CuSO(4)·5H(2)O (150 mg L(-1)) and MnSO(4)·H(2)O (150 mg L(-1)) in a separate cluster. These groups were linked with all of the other samples treated with metals at concentrations of 150 mg L(-1) and CuSO(4)·5H(2)O and Cd(NO(3))(2) at concentrations of 350 mg L(-1). Finally, the samples treated with metals at concentrations of 350 mg L(-1) together with NiSO(4) at the concentration of 150 mg L(-1), clustered separately. The DNA polymorphism detected by RAPD analysis offered a useful biomarker assay for the detection of toxic chemicals genotoxicity in plant model systems.

Assessment of heavy metal pollution in Republic of Macedonia using a plant assay.

Different plant organs (leaves, flowers, stems, or roots) from four plant species-Urtica dioica L. (Urticaceae), Robinia pseudoacacia L. (Fabaceae), Taraxacum officinale (Asteraceae), and Matricaria recutita (Asteraceae)-were evaluated as possible bioindicators of heavy-metal pollution in Republic of Macedonia. Concentrations of Pb, Cu, Cd, Mn, Ni, and Zn were determined in unwashed plant parts collected from areas with different degrees of metal pollution by ICP-AES. All these elements were found to be at high levels in samples collected from an industrial area. Maximum Pb concentration was 174.52 ± 1.04 mg kg⁻¹ in R. pseudoacacia flowers sampled from the Veles area, where lead and zinc metallurgical activities were present. In all control samples, the Cd concentrations were found to be under the limit of detection (LOD <0.1 mg kg⁻¹) except for R. pseudoacacia flowers and T. officinale roots. The maximum Cd concentration was 7.97 ± 0.15 mg kg⁻¹ in R. pseudoacacia flowers from the Veles area. Nickel concentrations were in the range from 1.90 ± 0.04 to 5.74 ± 0.03 mg kg⁻¹. For U. dioica leaves and R. pseudoacacia flowers sampled near a lead-smelting plant, concentrations of 465.0 ± 0.55 and 403.56 ± 0.34 mg kg⁻¹ Zn were detected, respectively. In all control samples, results for Zn were low, ranging from 10.2 ± 0.05 to 38.70 ± 0.18 mg kg⁻¹. In this study, it was found that the flower of R. pseudoacacia was a better bioindicator of heavy-metal pollution than other plant parts. Summarizing the results, it can be concluded that T. officinale, U. dioica, and R. pseudoacacia were better metal accumulators and M. recutita was a metal avoider.

Content of toxic and essential metals in medicinal herbs growing in polluted and unpolluted areas of Macedonia.

The aim of this study was to determine and compare Ba, Cr, Cd, Fe, Sr, Pb, and Zn content in medicinal herbs Urtica dioica L., Taraxacum officinale, and Matricaria recutita growing in polluted and unpolluted areas of the Republic of Macedonia. The metal content was determined using inductively coupled plasma atomic emission spectroscopy (ICP-AES). In the unpolluted area of Mt. Plackovica the metal content in Taraxacum officinale was in the descending order: Fe>Sr>Zn>Ba>Cr, while Pb and Cd were below the limit of detection. In the polluted area of Veles, the order was as follows: Fe>Zn>Sr>Pb>Ba>Cd>Cr. Our results suggest that quality assurance and monitoring of toxic metals is needed for plants intended for human use and consumption. Medicinal plants should be picked in areas free of any contamination sources.

In vitro antioxidant activity of some Teucrium species (Lamiaceae).

The chemical composition and antioxidant activity of different extracts (diethyl ether, ethyl acetate and n-butanol) obtained from Teucrium species (T. chamaedrys, T. montanum, T. polium) were investigated in this work. Phytochemical screening of the plant extracts proved the presence of flavonoids luteolin, apigenin and/or diosmetin. The chemical composition of extracts was evaluated by HPLC and spectrophotometry. Antioxidant activities of the extracts were evaluated using three complementary in vitro assays: inhibition of DPPH (1,1-diphenyl-2-picrylhydrazyl) radical, inhibition of hydroxyl radicals and protection of beta-carotene-linoleic acid model system. In the first two assays, strong inhibitory activity was shown by T. montanum and T. chamaedrys extracts. In the beta-carotene-linoleic acid model system, extracts from T. polium showed remarkable activity. These findings demonstrated that Teucrium species possess free radical and hydroxyl radical scavenging activity as well as antioxidant activity in vitro.