Vicia Faba Root Cells Research Articles

Genotoxic effects and mitosis aberrations of chromium (VI) on root cells of Vicia faba and its molecular docking analysis

Like other heavy metals, Cr (VI) is a powerful carcinogen and mutagen agent. Its toxic effects on plants are well considered. In order to elucidate its adverse effects, the present work aims to study the mitosis aberrations of Cr (VI) on the Vicia faba root-cells and its molecular docking analysis to understand the genotoxicity mechanisms. In-vivo, Vicia faba plants were exposed to 50 and 100 μM Cr (VI) for 48 h. In-silico, molecular docking and molecular dynamics simulation were used to study the interactions between dichromate and tubulin tyrosine ligase T2R-TTL (PDBID: 5XIW) with reference to Colchicine (microtubule inhibitor). According to our results, Cr (VI) affects growth and cell division and also induces many mitosis aberrations such as chromosome sticking, anaphase/telophase bridges, lagging chromosomes and fragmentation during all phases of mitosis. On the one hand, Cr (VI) reduces mitotic index and promotes micronuclei induction. The in-silico results showed that dichromate establishes very strong bonds at the binding site of the tubulin tyrosine ligase T2R-TTL, with a binding affinity of −5.17 Kcal/Mol and an inhibition constant of 163.59 μM. These interactions are similar to those of colchicine with this protein, so dichromate could be a very potent inhibitor of this protein's activity. TTL plays a fundamental role in the tyrosination/detyrosination of tubulin, which is crucial to the regulation of the microtubule cytoskeleton. Its inhibition leads to the appearance of many morphogenic abnormalities such as mitosis aberrations. In conclusion, our data confirm the highest genotoxicity effects of Cr (VI) on Vicia faba root-cells.

EVALUACIÓN DE LA GENOTOXICIDAD DE UNA SOPA INSTANTÁNEA COMERCIAL UTILIZANDO LA PRUEBA DE MICRONUCLEOS EN VICIA FABA Y RATÓN CD-1

Due to the high market demand that instant soup has among the Mexican population, the question arose whether it may cause any mutagenic effects after its consumption. The aim of this study was to analyze the cyto-genotoxic effects caused after the exposure of this food product compounds to meristemal root cells of Vicia faba and to peripheral blood cells of CD-1 mice. To determine this effect, it was performed both analysis of micronuclei as well as toxicity degree by using these two biological models. The interpretation of our statistical significance tests (ANOVA and Dunnett’s multiple comparison p<0.05), allowed us to conclude that nor the soup chemical compounds neither the packaging induce clastogenic damage but they cause a slight delay on cell division to V. faba; nevertheless the compounds can cause clastogenic damage and cytotoxicity to peripheral blood cells of CD-1 mice.

Do cancer cells in human and meristematic cells in plant exhibit similar responses toward plant extracts with cytotoxic activities?

We examined the effect of water extracts of Persea americana fruit, and of the leaves of Tabernamontana divericata, Nerium oleander and Annona cherimolia (positive control) on Vicia faba root cells. We had confirmed in our previously published data the cytotoxicity of these plant extracts on four human cancer cell lines: liver (HepG-2), lung (A549), colon (HT-29) and breast (MCF-7). Vicia faba roots were soaked in plant extracts at dilutions of 100, 1,250, 2,500, 5,000, 10,000, 20,000ppm for 4 and 24h. All treatments resulted in a significant reduction in the mitotic index in a dose dependant manner. Root cells treated with T. divericata, N. oleander and A. cherimolia exhibited a decrease in prophase cell percentage, increase in micronuclei and chromosomal abnormalities as concentration increased. The P. americana treatment showed the highest cytotoxic effect on cancer cells, prophase cell percentage increased linearly with the applied concentration and no micronuclei were detected. This study shows that root tip assay of beans can be used in initial screening for new plant extracts to validate their use as candidates for containing active cytotoxic agents against malignant cells. This will greatly help in exploring new plant extracts as drugs for cancer treatment.

Polycyclic Aromatic Hydrocarbons, Heavy Metals, and Genotoxicity of the Suburban Soils from Guangzhou, China

During the past few decades, urban and suburban developments have grown at unprecedented rates and extents with unknown consequences for ecosystem function. The problem of soil pollution as a result of the accelerating development of Guangzhou in China is becoming great concerns. In the present study, gas chromatograph coupled mass spectrometry (GC-MS), inductively coupled plasma mass spectrometry (ICP-MS) and inductively coupled plasma atomic emission spectrometry (ICP-AES) were employed to determine the 16 US Environmental Protection Agency (EPA) priority polycyclic aromatic hydrocarbons (PAHs) and the heavy metals (As, Cr, Cu, Pb, Cd, Hg, and Se) of soils collected from suburban areas of Guangzhou. The genotoxicity of these soils was screened with micronucleus (MN) assay in Vicia faba root cells. The concentrations of the pollutants in the soils were (dried weight): ΣPAHs (230.6–1263 ng·g−1), As (2282.6–36064 μg·kg−1), Cr (7109–64699 μg·kg−1), Cu (7047–56388 μg·kg−1), Pb (9675.9–93739 μg·kg−1), Cd (68.5–847.3 μg·kg−1), Hg (85.4–549.2 μg·kg−1), and Se (219.2–968 μg·kg−1), which fell in the moderately polluted range. However, six out of nine soil-exposed groups had a significant increases of MN frequencies observed in the V. faba root cells compared with the negative group (P < 0.05, P < 0.01), indicating that they had potential genotoxic risks. Bringing together the chemical analyses with the biological effects observed in this study, the genotoxic response could at a certain degree be explained by both the soil PAHs and heavy metals. Our results suggested that apart from chemical analysis, bioassays like the MN assay of V. faba root cells should also be included in a battery of tests to assess the eco-environmental risks of urban and/or urbanization in the developing areas on the soils.

Mutagenicity, Genotoxicity, and Estrogenic Activity of River Porewaters

We investigated mutagenicity, genotoxicity, and estrogenic activity in the porewaters of two river basins in southern Italy that had different features. Three samples from each site were collected in different seasons from 7 sites for a total of 21 samples. Mutagenicity was measured with the Ames test with and without metabolic activation (S9) using Salmonella typhimurium TA98 and TA100 strains. Genotoxicity was measured with two tests: one involved a chromophore that detected DNA damage in Escherichia coli PQ37 (SOS chromotest), and the other measured micronuclei formation in the root cells of Vicia faba. Estrogenic activity was measured with a yeast-based estrogen receptor assay and an MCF-7 cell-based, estrogen-sensitive proliferation assay. We also applied chemical analyses to detect alkylphenols, pesticides, natural and synthetic hormones, and heavy metals. The porewaters of both river sediments showed mutagenic/genotoxic activity on V. faba test and Ames test, the latter both with and without S9 liver fraction. The SOS chromotest without metabolic activation was not sufficiently sensitive to detect genotoxicity of the porewaters, but the SOS DNA repair system in E. coli PQ37 was activated in the presence of S9 mix. Good correlations were found between mutagenicity/genotoxicity and the concentration of cadmium and between estrogenic activity and the presence of copper. This study assessed the chemical concentrations of some bioavailable pollutants in porewater and detected the overall effects of multiple pollutants that contributed to mutagenicity, genotoxicity, and estrogenic activity of these two basin porewaters, thus increasing our understanding of the environmental consequences of polluted aquatic ecosystems.

Evaluation of the genotoxic potential of reactive black 5 solutions subjected to decolorizing treatments by three fungal strains

The genotoxic potential of solutions of the textile dye “Reactive Black 5” that were subjected to decolorizing treatments with the fungal strains Coriolopsis polyzona MUCL33483, Penicillium sp. MUBA001 and Pycnoporus sp. MUBA002 was tested. The genotoxicity of the solutions was determined by evaluation of micronuclei formation in Vicia faba root cells and calculation of a damage index (MNID). Non-treated Reactive Black 5 solutions (50–1000ppm) caused a statistically significant increase in micronuclei formation and, by then, in damage index. Solutions of dye treated with C. polyzona MUCL33483 and Pycnoporus sp. MUBA002 showed color loss, probably due to enzymatic breakdown of the colorant, but maintenance or even an increase in genotoxicity. On the other hand, the Penicillium sp. strain MUBA001 caused decolorization of the dye, apparently by adsorption on mycelia, and, for solutions that initially contained 50ppm of colorant, an elimination of the genotoxicity was observed after three weeks of treatment.

Lead-induced DNA damage in Vicia faba root cells: Potential involvement of oxidative stress

Genotoxic effects of lead (0–20μM) were investigated in whole-plant roots of Vicia faba L., grown hydroponically under controlled conditions. Lead-induced DNA damage in V. faba roots was evaluated by use of the comet assay, which allowed the detection of DNA strand-breakage and with the V. faba micronucleus test, which revealed chromosome aberrations. The results clearly indicate that lead induced DNA fragmentation in a dose-dependant manner with a maximum effect at 10μM. In addition, at this concentration, DNA damage time-dependently increased until 12h. Then, a decrease in DNA damages was recorded. The significant induction of micronucleus formation also reinforced the genotoxic character of this metal. Direct interaction of lead with DNA was also evaluated with the a-cellular comet assay. The data showed that DNA breakages were not associated with a direct effect of lead on DNA. In order to investigate the relationship between lead genotoxicity and oxidative stress, V. faba were exposed to lead in the presence or absence of the antioxidant Vitamin E, or the NADPH-oxidase inhibitor dephenylene iodonium (DPI). The total inhibition of the genotoxic effects of lead (DNA breakage and micronucleus formation) by these compounds reveals the major role of reactive oxygen species (ROS) in the genotoxicity of lead. These results highlight, for the first time in vivo and in whole-plant roots, the relationship between ROS, DNA strand-breaks and chromosome aberrations induced by lead.

Eco-toxicity and metal contamination of paddy soil in an e-wastes recycling area

Paddy soil samples taken from different sites in an old primitive electronic-waste (e-waste) processing region were examined for eco-toxicity and metal contamination. Using the environmental quality standard for soils (China, Grade II) as reference, soil samples of two sites were weakly contaminated with trace metal, but site G was heavily contaminated with Cd (6.37 mg kg −1), and weakly contaminated with Cu (256.36 mg kg −1) and Zn (209.85 mg kg −1). Zn appeared to be strongly bound in the residual fraction (72.24–77.86%), no matter the soil was metal contaminated or not. However, more than 9% Cd and 16% Cu was present in the non-residual fraction in the metal contaminated soils than in the uncontaminated soil, especially for site G and site F. Compared with that of the control soil, the micronucleus rates of site G and site F soil treatments increased by 2.7-fold and 1.7-fold, respectively. Low germination rates were observed in site C (50%) and site G (50%) soil extraction treated rice seeds. The shortest root length (0.2377 cm) was observed in site G soil treated groups, which is only 37.57% of that of the control soil treated groups. All of the micronucleus ratio of Vicia faba root cells, rice germination rate and root length after treatment of soil extraction indicate the eco-toxicity in site F and G soils although the three indexes are different in sensitivity to soil metal contamination.

In vitro evaluation of the anti-mutagenic effect of Origanum majorana extract on the meristemetic root cells of Vicia faba

The present investigation, the antimutagenic potential of Origanum majorana was evaluated in Vicia faba root meristematic cells. The root tip cells of Vicia faba were treated with sodium azide at 250 and 350 μg/ml for 6 h and Origanum majorana was given at 50, 100 and 200 μg/ml for 20 h, prior to sodium azide treatment. The tips were squashed after colchicine treatment and the cells were analyzed for chromosome aberration and mitotic index. Origanum majorana induces chromosomal aberration in Vicia faba root tip cells in an insignificant manner, when compared with untreated control. Sodium azide alone induces chromosomal aberrations significantly with increasing concentrations. The total number of aberrations was significantly reduced in root tip cells pretreated with Origanum majorana. This study revealed that Origanum majorana has antimutagenic potential against sodium azide induced chromosomal aberrations in Vicia faba root meristematic cells. In addition, it showed mild cytotoxicity by reducing the percentage of mitotic index in all O. majorana treated groups, The antimutagenic potential of Origanum majorana is effective at 50 μg/ml in Vicia faba root meristematic cells.

Effect of Static Electric Fields in Root Cells of Vicia faba (Fabaceae)

Serial electron microscopic sections were prepared from half-ripened meristematic root cells of Vicia faba (Fabaceae) which had been exposed gradually to 700, 1000, 2500, 3500, and 5000 V/m static electric fields during seven days with and without Zn and Cd electrodes. At the end of five weeks, wall loosenings and very small nuclei were observed in those root cells which were exposed to static electric currents from the lower side of the medium without electrodes, while abnormalities in cell formation, e.g., two cells with one nucleus, and GER occurrence were present in an electrolytic (Cd upward and Zn downward) medium. The cells exposed to a static current from the upper side of the medium had small nuclei and abnormal cell divisions in the electrolyte, but in a non-electrolyte very large nuclei and thicker cell walls were observed, the cytoplasm was dense with GER, pinocytosis was seen filled with mitochondria, and protoplast formation with big nuclei was seen in exocytosis.

Genotoxicity of drinking water disinfectants in plant bioassays

The genotoxicity of two widely used drinking water disinfectants, sodium hypochlorite (NaClO) and chlorine dioxide (ClO(2)), and a new disinfectant, peracetic acid (PAA, CH(3)-CO-COOH), was evaluated in three short-term plant tests: (1) induction of anaphase chromosome aberrations in the root cells of Allium cepa, (2) micronucleus induction in the root cells of Vicia faba, and (3) micronucleus induction in Tradescantia pollen cells. The study was carried out in the laboratory by directly exposing the plants to several concentrations of the disinfectants in redistilled water at unadjusted (acid) and adjusted (neutral) pHs. Both 0.1 and 0.2 mg/l NaClO induced chromosome aberrations in the Allium cepa test at acid pH, but concentrations up to 0.5 mg/l of all the disinfectants were negative at neutral pH. Concentrations ranging from 0.1 to 0.5 mg/l NaClO, ClO(2,) and PAA induced micronuclei in Vicia faba at acid pH, while 1-2 mg/l NaClO and ClO(2) and 0.5-2 mg/l PAA gave positive responses at neutral pH. Most of concentrations of ClO(2) produced positive responses in the Tradescantia micronucleus test. In general, the highest levels of genotoxicity were observed under acid conditions; at acid pH, significant effects were induced by low concentrations of ClO(2) and PAA. Since the test concentrations of disinfectants are typical of those encountered in the biocidal treatment of tap water and similar concentrations are consumed daily by a large number of people, the genotoxicity of these compounds may constitute a significant public health concern.

Genotoxicity of surface water treated with different disinfectants using in situ plant tests.

Disinfection of surface drinking water, in particular water chlorination, results in many by-products with potential genotoxic and/or carcinogenic activity. In the present study, we evaluated the genotoxicity of surface water after treatment with different disinfectants by means of in situ plant genotoxicity assays (micronucleus and chromosomal aberration tests) which can detect both clastogenic and aneugenic effects. The study was carried out at a pilot plant using lake water after sedimentation and filtration. This water supplied four stainless steel basins: three basins were disinfected with sodium hypochlorite, chlorine dioxide, and peracetic acid and the fourth basin containing untreated lake water was used as a control. Plants were exposed in situ in the basins. The study was carried out using water collected in different seasons over a period of about 1 year in order to assess the treatments in different physical and chemical lake water conditions. The micronucleus test in root cells of Vicia faba (Vicia faba/MCN test) revealed genotoxicity in many samples of disinfected water. The micronucleus test in Tradescantia pollen cells and the chromosome aberration test in root cells of Allium cepa showed genotoxic effects only in some disinfected samples, but also revealed genotoxicity in raw water. The results of the study indicated that the Vicia faba/MCN test was the most sensitive plant assay for disinfected water and that peracetic acid disinfection produced similar or lower genotoxicity than sodium hypochlorite or chlorine dioxide treatment.

Sister Chromatid Exchanges Induced by Heavy Metals in Vicia Faba

The induction of sister chromatid exchanges (SCE) by chloride and nitrate salts of nickel, cobalt, cadmium and zinc were studied in meristematic root cells of Vicia faba. Salts of nickel, cobalt and cadmium significantly increased the frequency of SCE, whereas chloride and nitrate salts of zinc did not increase the frequency of SCE significantly above the spontaneous level. The reported data demonstrate that the induction of SCE in Vicia faba may represent a valuable bioindicator for detecting the cytogenetic damage of heavy metals.

Arrest of Mitotic Cycle and Induction of Chromosomal Aberrations by Aflatoxin B2 in Root Cells of Vicia faba L.

Image cytometric measurements demonstrated a dose-dependent effect of aflatoxin B2 (AFB2) treatments on the components of mitotic cell cycle in Vicia faba L. root meristematic cells. The most evident effect appears to be the accumulation of cells in the G0/G1 phase at the expense of other phases of the cycle (S phase, G2 phase, M phase). These results indicate that this toxin acts as an inhibitor of cell cycle progression at the G1 transition point. In addition, a dose-dependent increase in a fraction of cells having <2C DNA and >4C DNA results from the AFB2 treatments. The inhibition of mitotic activity induced by AFB2 treatments is associated with a reduction in seedling growth. Cytological examination of dividing cells revealed an abundance of dose-dependent chromosome abnormalities produced by the applied treatments of AFB2. Chromosomal abnormalities associated with stickiness of chromosomes or due to an action on the mitotic apparatus are the dominant abnormalities induced by this toxin. Some of these abnormalities, particularly chromosome lagging and multipolar ana-telophase configurations, may account for the formation of cells with <the 2C DNA value or more >4C DNA value. However, true clastogenic chromosomal aberrations including chromosome breaks and ring chromosomes at metaphase and chromosomal bridges at ana-telophase were scored in substantial proportion of cells. The induction of whole chromosome breaks at metaphase is congruent with the indication by the cytometric measurements that AFB2 acts on the G, transition point. The capacity of this toxin to induce clastogenic aberrations may be regarded as an indication of its genotoxic potential. This is also indicated by the formation of micronuclei in interphase cells.

The alkaline comet test on plant cells: A new genotoxicity test for DNA strand breaks in Vicia faba root cells

The alkaline comet assay (single cell gel electrophoresis assay) is a sensitive method for the detection of DNA damage. This paper describes the first application of this assay to plant cells for genotoxicological assessment. Germinating Vicia faba (field bean) seedlings were kept in water with either methyl methanesulfonate (MMS), ethyl methanesulfonate (EMS), mitomycin C (MMC), cycloheximide (CH), cadmium chloride (CdCl 2), potassium dichromate (K 2Cr 2O 7), or chromium trichloride (CrCl 3). Nuclei were isolated from the root cells and evaluated for the extent of DNA migration. With the exception of cycloheximide, all agents induced a significant increase in DNA migration. These results indicate that the comet assay may be a valuable tool for monitoring DNA damage in plant systems. However, there was a significant heterogeneity in the extent of DNA migration within and between seedlings, which may be intrinsic to the assay or indicative of sampling problems.

Clastogenicity of lanthanides — induction of micronuclei in root tips of Vicia faba

The clastogenic property of two lanthanides, Praseodymium and Neodymium, was evaluated by scoring the frequencies of micronucleated cells in the interphase and chromosome/chromatid aberrations at ana-telophases in root tips of Vicia faba L. The significant increase in the frequency of micronucleated cells coupled with the increase in the frequency of chromosome/chromatid bridges and lagging chromosomes/chromatids suggest that these two are clastogenic for Vicia faba root cells.

Induction of chromosome damage by aniline and its C-hydroxylated metabolites in the root cells of Vicia faba

The induction of sister-chromatid exchanges (SCEs) and chromosome aberrations (CAs) by aniline hydrochloride (AH) and its C-hydroxylated metabolites, o-, m- and p-aminophenol, in the root cells of Vicia faba was examined. AH induced CAs, but not SCEs. AI1 the C-hydroxylated metabolites of aniline induced both SCEs and CAs. However, the treatment of cells with these metabolites at concentrations that did not cause significant increases in CAs resulted in significant increases in SCEs. These results seem to suggest that the substance that induced CAs in root cells treated with AH was not the C-hydroxylated metabolites of aniline.

Induction of chromosome damage by benzo[ a]pyrene, 2-aminofluorene and cyclophosphamide in the root cells of Vicia faba

The induction of sister-chromatid exchanges (SCEs) and chromosome aberrations (CAs) by benzo[ a]pyrene (BP), 2-aminofluorene (2-AF) and cyclophosphamide (CP) in the root cells of Vicia faba was examined. BP and 2-AF induced CAs, but not SCEs. CP induced both SCEs and CAs.

Underrepresentation of nuclear DNA sequences in differentiating root cells ofVicia faba

Data from cytological and biochemical analyses are presented on the behaviour of nuclear DNA during the differentiation ofVicia faba root cells. From the terminal 10.5 mm of the root, three segments were dissected by cutting transversely the root at 0.5 (segments I, meristematic cells), 4.5 (segment II, both meristematic and differentiating cells) and 10.5 mm (segment III, differentiating and/or differentiated cells) from the tip. Cytophotometric determinations of Feulgen absorptions in cell nuclei of the three root segments, carried out in preparations subjected to hydrolysis curve, revealed a lesser amount of nuclear DNA in differentiating cells when compared to the meristematic ones. Analyses of the reassociation kinetics of the DNAs extracted separately from the three root segments showed differences in the frequency of highly repeated sequences, which amount to 11.0, 8.6, and 7.5% of the total DNA in segments I, II, and III, respectively. Density gradient centrifugations in CsCl revealed a lighter satellite in the DNAs from segments I and II (ca. 5.4 and 3.8% of the total DNA, respectively) and no satellite in the DNA from segment III. It is suggested that underrepresentation of repeated DNA sequences occurs in differentiating cells and is a determining factor of the discharge of a cell from the mitotic activity.

Changes in the polyadenylated messenger RNA population during differentiation of Vicia faba root cells

Using cDNA probes, the polysomal polyadenylated RNA populations of Vicia faba meristematic, elongating, and mature root cells were analysed and compared, with respect to complexity, abundance distribution, and sequence representation. These cells express 12 300, 15 800, and 15 200 sequences, respectively, of an average size of 1300 nucleotides, distributed in three frequency classes. Transition from the meristematic to the elongating stage is coordinated with the disappearance of 25–30% of the abundant RNA species (2000 copies per cell) and with the appearance of new transcripts corresponding to 3000–3500 genes expressed at an intermediate level (50–60 copies per cell) and belonging to the rare class (about 4 copies per cell). These new transcripts represent 12.5% of the mass of the mRNA during the elongating stage and are quantitatively modulated during the transition to the mature stage. Thus 80 to 85% of the polysomal polyadenylated RNA species expressed in elongating or mature cells are common to the three cell types.