Plant Test Systems Research Articles

THE SUBSTANCE OF SELECTION OF PLANT TEST SYSTEMS FOR ESTIMATION OF TOXICITY OF SURFACES OF THE LONG TERRITORIES OF GRANITE CAREERS

The analysis of existing criteria for the selection of plant test systems is carried out in order to assess the impact of granite quarry dumps on the soil conditions of adjacent territories. The advantages and disadvantages of bioindicators plants most common in the territories of granite quarry concentration in Ukraine are determined. The hierarchy analysis method has been applied to justify the use of Onium (Allium cepa L.) and Crested Salad (Lepidium sativum L.) for assessing the ecological status of soils in the adjacent territories of granite quarries, which will provide complete and sufficient information not only about the pollution sources, but also to evaluate the impact of such pollution.

English

Considering the vast use of sweeteners by the Brazilian population and the constant need for toxicological studies of food additives, this study aimed to evaluate the cytotoxic and mutagenic potentials of the sodium saccharin and/or sodium cyclamate sweeteners in plant (Allium cepa) and animal (Mus musculus) test systems based on concentrations permitted by the Brazilian laws. In A. cepa, both the sweeteners individually and their combinations concentration and exposure time (ET: 48, 72 and 168 h) dependently exerted cytotoxic and mutagenic effects. Similarly, an increased in micronuclei formation was also observed by the sweeteners in peripheral blood cells in mice. More toxic effects were observed with the combination doses at 168 ET. There may be a synergistic effect and DNA damage with an increasing concentration and ET. In conclusion, the concentrations considered safe by the Brazilian laws had significant cytotoxic and mutagenic activity on the eukaryotic cells. Key words: Allium cepa, Mus musculus, sweeteners, toxicity.

Mutagenic potential of some heavy metals

Heavy metals are a drastically different group in terms of their mutagenic properties. The mutagenic potential of eight heavy metals (Ag, Cd, Cr, Cu, Hg, Mo, Pb, and W) has been studied using a Crepis capillaris L. plant test system. Ag and Cr have been found to be mutagens at the lowest concentrations (up to 10–8 M); Cu and W were not mutagens. A comparative analysis of the results obtained with other plant test systems was performed and close correlations were established. The toxic and mutagenic sequences of these metals were made up.

Toxic Effects of Gas Condensate on Soils

Oil and oil product pollution causes significant changes in the morphological and agronomic properties of soil. Plants can be considered the most convenient objects for soil biomonitoring. Simplicity of accounting effects and the interpretation of results makes appropriate use of plant test-systems for the diagnosis and assessment of toxicity of polluted soils. That is why the study of toxic effect of gas condensate on soils of different granulometric composition is necessary. The aim of the work is study of phytotoxicity of gas condensate of polluted soils in laboratory conditions. The authors have chosen gray and brown forest soils of Lviv region for investigation. The soil for research was collected from the root layer of natural areas at depths up to 20 cm. The uniform pollution of soil by different concentrations of gas condensate was simulated in laboratory conditions. The evaluation of soil phytotoxicity was conducted on the basis of test reactions of cress-salad: the length of the root and height of the stalk. Phytotoxic effect was calculated in percentage of the length of the root and height of the stalk. The results of the research revealed that gas condensate pollution of soils has a negative impact on growth of cress-salad stems. Inhibition of plant growth and development was proportion to the degree of pollution of gas condensate. Pollution concentration of 1-2 % has almost no effect on seed germination of cress-salad, but inhibits the development of shoots and roots. It is established that the gas condensate has a greater toxic effect on root length in brown forest soil and on the height of the stem in gray forest soil. Thus, gas condensate pollution of brown and gray forest soils inhibits the growth of root length and stem height of cress-salad. Gas condensate polluted soils, which differ in granulometric composition, have different phytotoxicity on growth parameters of root length and stem height cress-salad. Pollution concentration of 1-2 % gas condensate virtually no effect on seed germination cress-salad, but it inhibits the development of shoots and roots. It was researched that gas condensate has a greater toxic effect on root length in brown forest soil and on the height of the stem in gray forest soil.

Mutagenic potential of some heavy metals

Heavy metals are drastically different group from the point of view of its mutagenic properties. The mutagenic potential of eight heavy metals (Ag, Cd, Cr, Cu, Hg, Mo, Pb, W) was investigated using plant test-system Crepis capillaris L. Ag and Cr were mutagens at the lowest concentrations (up to 10-8M), Cu and W were not mutagens. Analysis of the results obtained with other plant test-systems was done and good correlations were established. Toxic and mutagenic sequences of these metals were made up.

Investigation on the effects of guava (Psidium guajava L.) infusions on germination, root tips and meristematic cells of Latuca sativa.

Guava (Psidium guajava L.) is a plant often employed in popular medicine. Recently several studies have alerted about the toxicity of substances present in medicinal plants, which can pose risks to the human health. In this sense, the present work aimed to investigate the phytotoxic, cytotoxic and genotoxic action of three guava varieties - Paluma, Pedro Sato and Roxa ("purple") - on the plant test system Lactuca sativa L. Thus, macro- and microscopic evaluations were carried out for five infusion concentrations (2.5, 5.0, 10.0, 20.0 and 40.0 g.L(-1)) prepared from each variety. Distilled water was used as negative control. Chromatographic and spectroscopic analysis by HPLC-PAD indicated that the chemical composition of the infusion of Roxa is different than that of the infusions of the varieties Paluma and Pedro Sato. It was observed that seed germination and root growth in L. sativa exposed to infusions decreased with increasing infusion concentration, regardless of the tested cultivar. For the mitotic index, no statistical differences were observed. On the other hand, a significant increase in the frequency of cell cycle alterations was verified, especially for the highest concentrations tested. The cytogenotoxic effect was significant. Therefore, guava should not be used indiscriminately in popular medicine.

Cytotoxicity of aqueous extracts of Rosmarinus officinalis L. (Labiatae) in plant test system.

This study investigated the cytotoxic activity of Rosmarinus officinalis L. (rosemary) aqueous extract on the cell cycle of Allium cepa. To this end, crude aqueous leaf extracts at four concentrations, 0.02, 0.04, 0.06 and 0.08 mg/mL, were tested on A. cepa meristematic root cells, at exposure times of 24 and 48 h. Slides were prepared by the crushing technique, and cells analyzed throughout the cell cycle, totaling 5,000 for each control group and concentration. The four concentrations tested, including the lowest and considered ideal for use, at all exposure times, showed a significant antiproliferative effect on the cell cycle of this test system and presented a high number of cells in prophase. Our results evidenced the cytotoxicity of rosemary extracts, under the studied conditions.

Protective Effects of Quercetin against Dimethoate-Induced Cytotoxicity and Genotoxicity in Allium sativum Test.

The present investigation was directed to study the possible protective activity of quercetin—a natural antioxidant against dimethoate-induced cyto- and genotoxicity in meristematic cells of Allium sativum. So far there is no report on the biological properties of quercetin in plant test systems. Chromosome breaks, multipolar anaphase, stick chromosome, and mitotic activity were undertaken in the current study as markers of cyto- and genotoxicity. Untreated control, quercetin controls (@ 5, 10 and 20 μg/mL for 3 h), and dimethoate exposed groups (@ 100 and 200 μg/mL for 3 h) were maintained. For protection against cytogenotoxicity, the root tip cells treated with dimethoate at 100 and 200 μg/mL for 3 h and quercetin treatment at 5, 10, and 20 μg/mL for 16 h, prior to dimethoate treatment, were undertaken. Quercetin was found to be neither cytotoxic nor genotoxic in Allium sativum control at these doses. A significant increase (P < 0.05) in chromosomal aberrations was noted in dimethoate treated Allium. Pretreatment of Allium sativum with quercetin significantly (P < 0.05) reduced dimethoate-induced genotoxicity and cytotoxicity in meristematic cells, and these effects were dose dependent. In conclusion, quercetin has a protective role in the abatement of dimethoate-induced cyto- and genotoxicity in the meristematic cells of Allium sativum that resides, at least in part, on its antioxidant effects.

Investigation of Sensitivity of the Allium cepa Test as an Alert System to Evaluate the Genotoxic Potential of Soil Contaminated by Heavy Metals

Soil is one of the compartments most affected by the accumulation of pollutants from anthropic sources. The present study allowed the identification of the sensitivity of the Allium cepa test system to evaluate solubilized soils from two points in the area contaminated by heavy metals, as well as a point of reference. They are all located in the municipality of Triunfo, state of Rio Grande do Sul, Brazil. The parameters used for evaluation were germination index, mitotic index, chromosomal aberrations (CA), and index of mutagenicity index (IMUT) presented by A. cepa. Significant responses of CA were observed in the two samples of contaminated soil, but IMUT was significant only for soil 3. The toxicity and cytotoxicity indexes did not show significant responses. The results indicate that the A. cepa plant test system was sensitive to investigate the genotoxicity of the soil samples and can be used as an alert in studies on soil contamination. It was partially concordant with the mutagenic responses already detected for the Salmonella/microsome assay in previous studies.

Genetic effects of root extracts of Glycyrrhiza glabra L. in different test systems

The antimutagenic and geroprotective activities of root extracts of Glycyrrhiza glabra have been demonstrated both on plant test systems--Allium fistulosum L., Allium cepa L., Vicia faba L. and on animals--Vistar rats. The possibilities of the mobilization of Glycyrrhiza glabra root extracts as antimutagenic agents are discussed.

Investigation of cytotoxic and mutagenic effects of Malpighia glabra L. (barbados cherry) fruit pulp and vitamin C on plant and animal test systems

Fruits are important sources of nutrients in human diet, and Barbados Cherry (Malpighia glabra L.) is of particular interest due to its high content of antioxidants. Diets rich in fruits and vegetables protect individuals against diseases and cancer, but excessive intake of vitamins may act as pro-oxidant and generate changes in DNA. To evaluate the effect of different in natura (BAN) and frozen (BAF) Barbados Cherry pulp concentrations and synthetic vitamin C in liquid form (VC) on the chromosome level and the cell cycle division, root meristeme cells of Allium cepa L. and bone marrow cells of Wistar rats Rattus norvegicus, were used as test system. In Allium cepa L., BAN, at the highest concentration (0.4 mg.mL-1) and BAF, at the lowest concentration (0.2 mg.mL-1), inhibited cell division, and there was recovery of cell division after the recovery period in water only for BAN. In the Wistar rats, all treatments with Barbados Cherry, either acute or subchronic, were not cytotoxic or mutagenic; only the highest concentration of VC increased significantly the rate of chromosomal abnormalities. The data obtained are important to reinforce the use of Barbados Cherry fruit in the diet.

About the action of metabolites of plant growth-promoting rhizobacteria Bacillus subtilis on plant salt tolerance (I)

To find out the mode of plant tolerance enhancement against salinity by plant growth-promoting rhizobacteria Bacillus subtilis, metabolites of strains FZB24 and FZB41 were studied in a test system with tomatoes under the influence of high salinity. The culture filtrate (CF) from the fermentative transitional phase, containing the whole range of produced metabolites by B. subtilis, showed to a certain extent tolerance-increasing action at dilution of 0.1% in the test plants with the parameters length, fresh mass and dry mass of shoots and roots as well as leaf area after 7-day treatment and subsequent plant cultivation under high salt stress. Afterwards, the CF was fractionated with adsorber resin and high performance liquid chromatography, and these fractions, as well as fractions from a CF after 19-h fermentation, were also tested with axenic-cultivated tomato seedlings. Fractions with different proteins and peptides, produced by B. subtilis, showed partly activities depending on concentration with regard to plant growth stimulation, including tolerance enhancement against salt stress. Subsequently, also an extract from B. subtilis culture with special concentrated peptides was examined in the axenic plant test system and showed similar activity depending on concentration. The observed effect of the bacterial metabolites is discussed as one part of the mechanism for plant growth stimulation and at the same time salt tolerance, increasing action of the rhizobacterium by its root colonization and interaction with the plant metabolism.

Multi-walled carbon nanotubes (MWCNT): Induction of DNA damage in plant and mammalian cells

Increasing use of multiwalled carbon nanotubes (MWCNT) necessitates an improved understanding of their potential impact on environment health. In the present study we evaluated the genotoxicity of MWCNT on plant and mammalian test systems. Genotoxic responses such as chromosomal aberrations and DNA strand breakages were studied in Allium cepa, human lymphocytes, mouse bone marrow cells and pBR322 plasmid DNA. Results showed that MWCNT could cause chromosomal aberrations, DNA fragmentation and apoptosis in Allium root cells that could be correlated with the internalization of MWCNT in the plant cells. In human lymphocytes significant genotoxic response was observed at the concentration 2μg/ml. Higher concentrations led to a decrease in values of the tail DNA percent that may be due to the formation of crosslinks. Annexin V-FITC–PI staining indicated only a small percentage of cells were undergoing apoptosis. Genotoxic effects were shown by micronuclei (MN) frequencies in experiments on mouse bone marrow cells. In the cell free DNA system (plasmid pBR322), a strong correlation between DNA strand break and concentration was observed. Based on the findings of the present study MWCNT may have significant impact on genomic activities.

Можливість використання рослинних тест-систем для біомоніторингу нафтозабруднених ґрунтів

Problems and perspectives of technogenic soils pollution have been analyzed. the Potentials of using plant test systems for biomonitoring oil polluted soils based on literature and authors data are evaluated. Clarification of these issues is a theoretical basis for interpreting the results of ecological and toxicological monitoring of oil polluted areas. Keywords: phytotoxicity, oil polluted soils, plants test systems, biomonitoring, biotesting.

Chromosome aberrations induced by curcumin and aloin in Allium cepa L. root meristem cells

To date, a variety of plant compounds have been studied extensively with regard to anti-mutagenic, anti-carcinogenic, and anti-genotoxic activity. Nevertheless, very few of them have been studied in terms of clastogenicity. In the present study the clastogenic activity of curcumin (the active component of turmeric) and aloin (the active component of aloe) was evaluated at doses of 5, 10, 20, 30, 50, and 100 µg/mL for 4, 8, and 12 h. Both curcumin and aloin had significant dose- and time-dependent clastogenic effects on the test plant system (Allium cepa L.). Compared to curcumin, aloin was more clastogenic. Clastogenic activity was obvious at very low concentrations of curcumin and aloin at all time points. The mitotic index was significantly lower at higher concentrations; however, the exact mechanism of curcumin and aloin clastogenicity remains unknown in plant test systems.

Antimitotic effects of the biopesticide oxymatrine

Abstract Biopesticides offer a more sustainable solution to pest control than synthetic alternatives. Naturally plant products, called secondary metabolites include thousands of alkaloids, terpenoids, phenolics and minor secondary chemicals. Sophora flavescens Aiton (Leguminosae) is an ancient Chinese herb that grows wild in China. About two dozen alkaloids have been identified in Ku Shen, with matrine and oxymatrine being the major compounds. Ku Shen extracts have been formulated as pesticides either alone or in mixtures with conventional synthetic pesticides. In this study we have analyzed the probable cytotoxic and genotoxic effects of oxymatrine on plant test system for determining the safety of use as biopesticide. This alkaloid has reduced the mitotic index in Allium root meristem in higher concentrations but hasn’t show distinct genotoxic effects. In conclusion oxymatrine can be proposed as safe at this end point for using as biopesticide while the plant physiological system can ruled out certain e...

Avaliação da atividade mutagênica e antimutagênica da Aloe vera em teste de Allium cepa e teste de micronúcleo em linfócitos humanos binucleados

Algumas das substâncias presentes nos vegetais podem ter efeitos mutagênicos, enquanto outras podem atenuar ou anular estes efeitos. Estima-se que mais de 200 substâncias ativas façam parte do gel de Aloe vera, sendo que os polissacarídeos chegam a 30%, e muitos dos benefícios terapêuticos, nutricionais e cosméticos são a eles atribuídos. O objetivo deste trabalho foi determinar o efeito mutagênico e antimutagênico da solução de Aloe vera, em teste de Allium cepa e teste de micronúcleos em linfócitos humanos binucleados. Para as soluções, as folhas de Aloe vera foram trituradas, filtradas e esterilizadas. O teste de Allium cepa foi constituído de oito tratamentos e o de linfócitos, de cinco (com quatro repetições). Observou-se que na dose usual, a solução de Aloe vera não foi mutagência para o sistema de teste vegetal e nem para o humano. Já na dose dez vezes mais concentrada provocou um efeito citotóxico e mutagênico em Allium cepa. Em células vegetais a solução foi antimutagênica apenas quando colocada depois do paracetamol, enquanto que nas células humanas, essa ação se manifestou quando a solução foi utilizada ao mesmo tempo com o paracetamol.

Effect of the aqueous extract of Banana Fruits Peal Musa paradisiaca on Mitosis in Plant and Mammalian cells

The aqueous extract of banana fruits peal was tested for its effect on mitosis . The root tips of Allium cepa were used as plant test system and the bone marrow cells of the albino mice Mus musculus were used as mammalians test system in vivo .Root tips of Allium cepa were treated for four hours with five concentrations of the extract (5 , 10 , 20 , 40 ,60 mg / ml.).The Metaphase was arrested in all the treatments , the highest percentage ( 100 % ) was recorded in the first concentration , the last concentration caused stickiness and clumping of the chromosomes. The treatments did not cause significant difference in the mitotic index. The peals extract (5 mg /ml) was compared with the extracts of fruits bulb, leaves and roots of banana plant, it was found that the extract of fruits peal is the best considering the highest percentages of arrested Metaphase in the root tips cells. The albino mice Mus musculus were injected intraperitonial with the peals extract ( 0.01 , 0.02, 0.04, 0.06, 0.08 mg / gm body weight), the percentages of arrested Metaphase in the bone marrow of these animals were comparable to the recorded percentages when the animals were injected with colchicine ( 0.01 mg / gm b.w.) .This study revealed the antimitotic activity of the aqueous extract of banana fruits peal on both the plant and mammalian cells in vivo. Studies will be conducted to investigate the effect of the extract and its components on the proliferation of cancer cells in vitro and in vivo.

Effect of the aqueous extract of Banana Fruits Peal Musa paradisiaca on Mitosis in Plant and Mammalian cells

The aqueous extract of banana fruits peal was tested for its effect on mitosis . The root tips of Allium cepa were used as plant test system and the bone marrow cells of the albino mice Mus musculus were used as mammalians test system in vivo .Root tips of Allium cepa were treated for four hours with five concentrations of the extract (5 , 10 , 20 , 40 ,60 mg / ml.).The Metaphase was arrested in all the treatments , the highest percentage ( 100 % ) was recorded in the first concentration , the last concentration caused stickiness and clumping of the chromosomes. The treatments did not cause significant difference in the mitotic index. The peals extract (5 mg /ml) was compared with the extracts of fruits bulb, leaves and roots of banana plant, it was found that the extract of fruits peal is the best considering the highest percentages of arrested Metaphase in the root tips cells. The albino mice Mus musculus were injected intraperitonial with the peals extract ( 0.01 , 0.02, 0.04, 0.06, 0.08 mg / gm body weight), the percentages of arrested Metaphase in the bone marrow of these animals were comparable to the recorded percentages when the animals were injected with colchicine ( 0.01 mg / gm b.w.) .This study revealed the antimitotic activity of the aqueous extract of banana fruits peal on both the plant and mammalian cells in vivo. Studies will be conducted to investigate the effect of the extract and its components on the proliferation of cancer cells in vitro and in vivo.

Effect of gamma‐sterilization and autoclaving on soil organic matter structure as studied by solid state NMR, UV and fluorescence spectroscopy

SummarySterilized soil is often used, for example in degradation studies, sorption experiments, microbiological tests and plant test systems, to distinguish between microbial processes and abiotic reactions. The most commonly used technique for sterilization is autoclaving of the soil. Another technique is irradiation with high‐level gamma radiation (γ‐radiation). One major drawback of sterilization procedures is the possible alteration of the structure of soil components, for example the organic matter. A change in the chemical structure of the soil organic matter can cause different reactions in the above‐mentioned experiments and hence interfere with the aim of clearly distinguishing between biotic and abiotic processes. Two soils (Gleyic Cambisol and Orthic Luvisol) were sterilized by two γ‐irradiation procedures (4 kGy hour−1 for 9 hours and 1.3 kGy hour−1 for 27 hours) and repeated autoclaving at 121°C. Gentle physical aggregate fractionation of the sterilized soils revealed a decrease in the aggregation of the soil, which was reflected in an increase of the clay fraction. Subsequent analysis of the aqueous phase revealed much more dissolved organic matter (DOM) in the γ‐sterilized and autoclaved soils than in the untreated soils. Ultraviolet (UV) and fluorescence spectra of the DOM showed a decrease in the aromaticity and polycondensation of the dissolved organic carbon (DOC). 13C cross‐polarization/magic‐angle spinning nuclear magnetic resonance (13C‐CP/MAS NMR) spectra of the unfractionated soils and their respective soil fractions before and after sterilization showed that the most important change occurred in the carbohydrate and N‐alkyl region, the main components of microorganisms. In general, the impact of the sterilization method was stronger for autoclaving. The γ‐sterilized soils and fractions displayed both fewer and smaller changes in the soil organic matter.