The plant Ranunculus acris (L.) is widespread. It is used in folk medicine. The use of herbal medicines can have adverse effects on humans and requires studies of herbal raw materials for ...genotoxicity. The goals of the study were to examine the toxic and genotoxic effects of aqueous solutions of R. acris flowers at concentrations of 0.625%, 1.25%, 2.5%, 5% and 10% using the Allium test and to determine the numbers of flavonoids and polyphenols and the antioxidant activity of the extracts. It has been shown that aqueous solutions of buttercup flower extract have a depressing effect on mitosis in the cells of the root meristem of Allium cepa (L.) and cause chromosomal abnormalities (chromosomal stickiness, chromosome lagging, nuclear buds and bridges in anaphase).
A general report on the use of the Allium test as cytotoxicological and genotoxicological assay is proposed, with particular emphasis about the standardization of the test in several common ...applications. The intraspecific variation in Allium cepa has been overlooked, as in most investigations no mention is made about origin and denomination of the onion cultivar used. A standardization of the used material would allow a better generalization of the results, since we cannot be sure that all cultivars would give the same response. A more frequent use of transmission electron microscopy (TEM) investigation is proposed. Even if relatively time consuming and not available in all laboratories, it may help to better understand the mechanism of cytotoxicity, since many morphological characters may appear similar but be arisen from different processes observable only with TEM. About statistical testing, tests other than chi-squared may be used in case of a lower amount of data. The most commonly used statistical tests are the parametric tests ANOVA and Student's t, and the non-parametric tests Kruskal-Wallis and Mann-Whitney U, for analysis of variance. Tests should be used also to assess the minimal sample dimension for obtaining significance, since data collection (microscope observation) appears to be one of the main bottle necks of the test. Also the use of the Allium test for testing liposomes and other nanovectors for drug delivery is proposed, in order to assess the cytotoxicity of these types of medium and the possible increase in cytotoxicity of the associated drug.
Triphenyl phosphate (TPhP) is a tri-ester of phosphoric acid and phenol. It is used in products such as polyvinyl chloride, hydraulic fluids, polymers, photographic films and printed circuit cards as ...a flame retardant. It is also widely used in consumer products like electric and electronic devices, textiles and building necessaries. TPhP can diffuse into its surrounding environment easily, because it does not form a chemical bond with polymeric materials. Therefore, bio-monitoring of such compounds is needed for possible environmental and human health effects. In this study, we evaluated genotoxicity and cytotoxicity of TPhP on Allium cepa on the cells of root tips. A 10 mg/mL stock solution of TPhP was dissolved in DMSO and dilutions of 2, 4, 6, 8 and 10 mg/mL was made in distilled water. Onion bulbs, rooted in test tubes, were exposed to these concentrations of TPhP for 24 h. Distilled water was used as a negative control, 0.7% hydrogen peroxide was used as positive control, and 0.5% DMSO was used as solvent control. Significant inhibition of onion root growth was observed following treatment with the 6, 8 and 10 mg/mL TPhP ranges in comparison with the negative and solvent control groups (p < 0.05). Furthermore, in the TPhP treatment groups, total chromosome aberration ratios were significantly high in comparison with the controls (p < 0.05). These results suggest that TPhP have cytotoxic and genotoxic effects on A. cepa root tips.
Background. Environmental contamination with radioactive materials of geogenic and anthropogenic origin is a global problem. A variety of mutagenicity test procedures has been developed which enable ...the detection of DNA damage caused by ionizing radiation. In the present study, we investigated the Allium test (the widely used plant-based genotoxicity bioassay) for the detection of genetic damage caused y-radiation. Received data indicate that this method is can be used for environmental biomonitoring. Aim of the present study was the investigation of y-radiation of the Allium test assay. Materials and methods. The bulbs of the Stuttgarter Riesen onion (Allium cepa L., 2n = 16) were kept in a cool dark place (+4–5 °C). For each sample and control were taken 3 bulbs. Distilled water was used as a control. Y - irradiation of the bulbs was carried out with the Eu152 for 3 days in doses of 0.3, 1 and 3 Gy. After 3 days, the roots were cut and fixed in Clark’s and to be placed in sealable test-tubes for long-time storing in 80% ethanol. The roots were simultaneously hydrolyzed and colored in ceramic crucibles in acetoorcein solution were boiled in the flame of an alcohol lamp. The slides prepared from squashing the root meristems with one drop of 45% acetic acid. The number of dividing cells was determined in 1000 examined cells in the field of view, with phase and chromosome aberration. Results. The appearance of giant cells, C-mitoses, nuclear buds, fragments, lagging and bridges, disturbances in the work of the fission spindle and micronuclei in irradiation at doses of 3 and 1 Gy were noted. Conclusion. As a result of the study, it was shown that y - irradiation of the onion test object (Allium cepa L.) with doses of 3, 1 and 0.3 Gy led to the appearance of cytogenetic disorders, the proportion of which significantly differed from the control.
Data from the present study indicate that the neonicotinoid insecticides Nuprid 200 SL (with the active substance imidacloprid) and Calypso 480 SC (with the active substance thiacloprid) reduce the ...mitotic activity by inhibiting cell division and increasing chromosomal aberrations in the cells of the root apical meristem of Allium cepa L. The tested solutions of pesticides cause a wide range of anomalies associated with disorders in forming the cell's division apparatus and the integrity of chromosomes. The comparative analysis of their mutagenic action shows a higher genotoxic potential of Calypso 480 SC. The established indices of chromosomal abnormalities for Calypso 480 SC and Nuprid 200 SL are 1.07 ± 0.38% and 0.70 ± 0.41% respectively. The mitosodepressant effect and a large number of chromosomal aberrations and mitotic abnormalities in Allium cepa cells when treated with Nuprid 200 SL and Calypso 480 SC are evidence of their high toxic potential and the significant risk of environmental pollution by their use in agriculture. Parts of this work was published in abstract form and presented as an oral presentation at the IV International Agricultural, Biological & Life Science conference, Edirne, Turkey, 2022 August 29-31, 1 and as a poster presentation at the 5th Balkan Scientific Conference on Biology, Plovdiv, Bulgaria, 2021 April 15-16 2.
The objective of this study was to explore the mutagenic and cytotoxic effects of Napoleon 4EC pesticide used in Turkey to control insect pest by using two standard assays. The Allium cepa test was ...used for determined the cytotoxic effects of this pesticide. For this test, onion seeds were exposed to Napoleon 4EC (100, 200, and 400 ppm) for 24, 48, and 72 hours. For each test group root tip cells were stained with Feulgen and five slides were prepared for each concentration and counted microscopically. The concentrations Napoleon 4EC was compared with the value for the negative control using Dunnet-t test, 2 sided. The results indicated that mitotic index was clearly decreased with increasing the concentration of Napoleon 4EC in each treatment group as compared to the controls. The percentage of mitotic phases has been markedly impacted. Five different doses of the pesticide (50, 100, 200, 400, 800 μg/plate) were examined with Ames test using Salmonella typhimurium strains TA98 and TA100 with and without S9 metabolic activation for mutagenic activity. Ames test results showed a dose dependent effect, but not twice the negative control for S. typhimurium TA98 and TA100, with or without S9 mix except 800 μg/plate doses. In 800 μg/plate doses, colony numbers are two-fold increase according to colony number of control group. So, this places the this compound as a weak mutagen according to the parameters.
Having started in the 1930-40s, the industrial development of the Kola North has caused elevated environmental pollution of the area’s water bodies. The pollutants contained in dust emissions, ...dissolved substances and waste waters require their genotoxicity to be estimated using in vivo assays. This article addresses the effect of pollutants entering the water bodies of the Murmansk region together with mining waste, which leads to a decrease in mitotic activity and an increase in chromosomal abnormalities in the roots of Allium cepa L. The evaluated waters showed an effect of reducing the mitotic index and the appearance of chromosomal aberrations; this may be associated with the presence of compounds in the water, such as mining waste.
Origanum vulgare L. has been proven to be the strongest herbal antiseptic in the world, native to the Mediterranean region, but is widely naturalized elsewhere in the temperate Northern Hemisphere. ...This study aimed to estimate the phytotoxic effect of three different concentrations of oregano essential oil (O. vulgare) on three selected plant species namely, wheat, tomato and mint using biotest germination and effects on seedling growth, as well as its toxicological properties using Allium test. Our results revealed that oregano essential oil exhibits allelopathic effect on selected species. All three tested concentrations of oregano essential oil caused a significant inhibition of Allium cepa L. root growth, as well as a reduction in the mitotic index values in A. cepa meristem cells. O. vulgare essential oil demonstrated phytotoxic and antiproliferative effects. Further research is needed to confirm our results.
Background. Bottom sediments (BS) of the Yenisei River contain xenobiotics of the chemical and radiation origins, which may adversely affect life functions of aquatic organisms. Previous assays of BS ...samples using the Allium test showed contradictory responses of the endpoints of onion bulbs (stimulation and inhibition) to the chemical and radionuclide composition of the BS. The Allium test with germinated seeds has not been used so far to test samples of the Yenisei BS, although previous studies showed sensitivity of this test object to exposure to low-dose gamma radiation. Purpose. The purpose of the study was to assess the possibility of using two modifications of the Allium test (A. cepa bulbs and seeds) to evaluate toxicity of samples of the Yenisei River BS containing relatively high activity concentrations of artificial radionuclides. Material and Methods. Toxicological laboratory experiments were conducted using three samples of the Yenisei BS containing high activity concentrations of the artificial radionuclide 137Cs; artificial BS prepared according to the standard protocol were used as the control. Activity concentrations of BS radionuclides were measured using a gamma-spectrometer coupled to a hyper-pure germanium detector (Canberra, U.S.). The test object was onion Allium cepa L. cv. Stuttgarter Riesen (bulbs and seeds). Root growth was chosen as the endpoint to determine BS toxicity. Results. Experiments with radioactive BS from the Yenisei River demonstrated significant stimulation of root growth in the modification with onion bulbs and inhibition of root growth in the modification with onion seeds. In previous research, stimulation of root growth was observed in onion bulbs exposed to Yenisei BS containing lower activity concentrations of 137Cs. For the first time, inhibition of root growth of onion seedlings was observed in assays with BS containing a wide range of 137Cs activity concentrations. Conclusion. Comparison of two modifications of the Allium test (bulbs and seeds) for assessing the toxicity of BS showed that only the growth of onion seedling roots used as an endpoint has sufficient sensitivity to the content of the artificial radionuclide 137Cs in BS and makes it possible to evaluate the biological effects of radioactivity. Funding. The study was funded by State Assignment of the Ministry of Science and Higher Education of the Russian Federation (project No. 0287-2021-0019).
Lanthanum (La) and cerium (Ce) are one of the most abundant rare earth elements (REEs). In spite of quite extensive studying of the effects of these lanthanides on biota, some contradictions remain ...in the results. Also little is known about the effect of lanthanum and cerium on plant cells and their mitotic cycle, especially in soils. In this study, the effects of La and Ce in solutions and soil samples on root growth, mitotic index (MI) and frequency of aberrant cells (FAC) were assayed using one of the most convenient objects for testing of cytotoxicity – onion Allium cepa L. Bulbs were germinated on media containing La and Ce in concentrations 0–200 mg/l and 0–50 mg/l respectively for solutions and 0–200 mg/kg for soil samples. After 5 days of germination in solutions, a significant decrease in root elongation and MI in apical meristem cells are shown. We have also observed an increase in the number of cells with aberrations at 50 mg/l La and Ce concentration. The number of observed stickiness and disturbed metaphase has increased significantly. Soil samples turned out to be less toxic compared to the solutions probably due to the decreased availability of REEs. In spite of this, significant cytotoxicity of soil samples containing the highest concentration of La and Ce (200 mg/kg) is observed. The latter may indicate the importance of considering the cytotoxicity of soils containing high lanthanides concentrations – in extraction and production areas and actively fertilized fields.
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•La and Ce inhibit cell division in onion root tips.•La and Ce increase the frequency of aberrant cells.•Cytotoxic effects of Ce is observed at lower concentration compared to La.•Toxic effects of La and Ce are less pronounced in the soil than in the solution.•La and Ce cause the disturbed metaphase and chromosome stickiness.
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