Vicia Faba Root Tip Cells Research Articles

Cytotoxicity and genotoxicity evaluation of chloroform using Vicia faba roots.

Chloroform is a widely used industrial chemical that can also pollute the environment. The aims of this study were to examine the potential cytotoxicity and genotoxicity of chloroform on plant cells, using the Vicia faba bioassay. Chloroform was evaluated at concentrations of 0.1, 0.5, 1, 2, and 5mg·L-1. The following parameters were analyzed: the mitotic index (MI), micronucleus (MN) frequency, chromosomal aberration (CA) frequency, and malondialdehyde (MDA) content. The results showed that exposure to increasing concentrations of chloroform caused a decrease in MI and an increase in the frequency of MN in Vicia faba root tip cells, relative to their controls. Moreover, various types of CA, including C-mitosis, fragments, bridges, laggard chromosomes, and multipolar mitosis, were observed in the treated cells. The frequency of MN was positively correlated with the frequency of CA in exposure to 0.1-1mg·L-1 chloroform. Furthermore, chloroform exposure induced membrane lipid peroxidation damage in the Vicia faba radicle, and a linear correlation was observed between the MDA content and the frequency of MN or CA. These findings indicated that chloroform exposure can result in oxidative stress, cytotoxicity, and genotoxicity in plant cells.

Preliminary assessment of genotoxic effects induced by radiation from EAST using Vicia faba micronucleus assay

During long-pulse deuterium plasma operations in the Experimental Advanced Superconducting Tokamak (EAST), a mixed radiation field is generated, which is mainly composed of fusion neutrons, gamma rays, and x-rays. More accurate and effective dose monitoring methods have been developed and established to determine the ionizing radiation intensity both for the stable operation of the device and for the radiation safety of personnel. As far as we know, there are few reports about the biological effects of radiation induced by fusion neutrons and γ radiation, which are of vital importance for the assessment of radiation hazards presented by fusion devices, such as EAST, to human beings and the environment. In this study, three positions in the EAST hall were selected to detect genotoxic effects induced by nuclear fusion radiation using a Vicia faba micronucleus (MN) test for the first time. The doses of neutrons and gamma rays at these places were measured by thermoluminescence dosimeters four times between June 2019 and May 2020. The radiation doses decreased as the distances from the EAST device shell gradually increased from S1 to S3. The radiation in the EAST hall resulted in a significant induction of MN in the Vicia faba root tip cells compared to a negative control, which was different from the MN frequency induced by fission neutrons, γ-rays and other kinds of radiation in previous studies. These results indicate the existence of potential genotoxic effects induced by radiation from EAST which is different from other radiation and suggest that personnel should not be permitted to enter the experimental hall during the discharge process, and that radiation protection measures should be taken during necessary maintenance to avoid radiation damage. These newly acquired results will certainly increase our knowledge about the biological effects induced by radiation from nuclear fusion and provide good data support for developing more effective environmental and personnel fusion radiation protection.

Immobilization of heavy metals in vegetable-growing soils using nano zero-valent iron modified attapulgite clay

Nowadays, the problem of heavy metal pollution in vegetables is received wide attention. In this work, attapulgite clay (ATTP), as a cheap and readily available inorganic mineral material, was modified with nano zero-valent iron (nFe0@ATTP) for heavy metal immobilization in soil. Batch experiments were employed to evaluate the optimal remediation performance by ATTP before and after modified with nFe0 through planting Pakchoi (Brassica chinesis L.) in Cd, Cr, and Pb contaminated soil from Changsha. The results showed that amendments can all increase the pH value of soils, and notably decrease the concentration of extractable Cd, Cr, and Pb in soil. The germination rate and root length of Pakchoi were promoted, and the activities of peroxidase (POD), catalase (CAT), and malondialdehyde (MDA) contents were notably reduced besides superoxide dismutase (SOD) activity after treatments with ATTP and nFe0@ATTP. Vicia faba-micronucleus test indicated that the application of amendments reduced the toxicity of heavy metals on the genetic material of Vicia faba root tip cells. The nFe0@ATTP were found to well convert Cd, Cr, and Pb into less bioavailable state in soil, thus blocking heavy metal uptake by plants. This material could be a promising amendment for heavy metals contaminated soil.

Influence of isolated bacterial strains on the in situ biodegradation of endosulfan and the reduction of endosulfan- contaminated soil toxicity

The recently discovered endosulfan-degrading bacterial strains Pusillimonas sp. JW2 and Bordetella petrii NS were isolated from endosulfan-polluted water and soil environments. The optimal conditions for the growth and biodegradation activity of the strains JW2 and NS were studied in detail. In addition, the ability of the strains JW2 and NS to biodegrade endosulfan in soils during in situ bioremediation experiments was investigated. At a concentration of 2 mg of endosulfan per kilogram of soil, both JW2 and NS had positive effects on the degradation of endosulfan; JW2 degraded 100% and 91.5% of α- and β-endosulfan, respectively, and NS degraded 95.1% and 90.3% of α- and β-endosulfan, respectively. Polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) of soil samples showed the successful colonization of JW2 and NS, and the toxicity of the soil decreased, as determined by single-cell gel electrophoresis (SCGE) assays of Eiseniafetida and micronucleus (MN) assays of Viciafaba root tip cells. Furthermore, the metabolic products of the bacterially degraded endosulfan from the in situ experiments were identified as endosulfan ether and lactone. This study provided potentially foundational backgrounds information for the remediation of endosulfan-contaminated soil.

Effect of the Water Extracts of Avocado Fruit and Cherimoya Leaf on Four Human Cancer Cell Lines and Vicia Faba Root Tip Cells

The main objective of this study is to determine the effect of the water extract of Persea americana Mill. (avocado fruit) and Annona cherimolia Mill (cherimoya leaf) on living cells. The antiproliferative properties of avocado fruit water extract (AFWE) and cherimoya leaf water extract (CLWE) were determined using four human cancer cell lines: lung (A549), liver (HepG-2), colon (HT-29) and breast (MCF-7). Cancer cells were incubated with 100 µg/ml of AFWE or CLWE for 48 hours, then the cell viability was measured using colorimetric tetrazolium cleavage test (MTT). Both extracts resulted in more than 90% mortality in treated cells. Vicia faba root tip assay was used to determine the effect of AFWE and CLWE on mitotic index (MI), Micronuclei (MN) formation rate and chromosomal abnormalities. Vicia faba roots were soaked in 100, 1250, 2500, 5000, 10000, 20000 µg/ml of AFWE or ALWE for 4 and 24 h. AFWE and CLWE were mitodepressive and resulted in a significant decrease of MI in a dose dependant manner. CLWE treatment resulted in a decrease in prophase cell percentage and an increase in MN & chromosomal abnormalities. On contrary, the prophase cell percentage was linearly increasing with the applied concentration without micronuclei formation in avocado treatment. Our results strongly indicate that avocado and cherimoya extracts were highly cytotoxic and mitodepressive on cancer and plant cells, respectively.

Impacts of nitrogen and phosphorus on atrazine-contaminated soil remediation and detoxification by Arthrobacter sp. strain HB-5

The relationship of atrazine-degrading bacteria Arthrobacter sp. HB-5 and nitrogen and phosphorus fertilizer to atrazine degradation and detoxification in soil was investigated in a microcosm pot experiment. Treatments of soil containing atrazine (AW) with atrazine plus strain HB-5 alone (A), together with atrazine and strain HB-5 plus nitrogen alone (AN), phosphate alone (AP), and nitrogen and phosphate together (ANP) were used to investigate atrazine degradation and ecotoxicity. Atrazine residues in the soils were determined by high performance liquid chromatography, while soil ecotoxicity was tested by micronucleus (MN) assay of Vicia faba root tip cells. The results showed that degradation of atrazine in soil could be facilitated by the treatment of strain HB-5 as well as strain HB-5 application with the addition of nitrogenous and/or phosphorus fertilizers. The degradation rates varied as the following: ANP > AP > AN > A > AW in different treatments. At 10 days post treatment, degradation efficiency of over 90 % was achieved in all strain HB-5 treatments except AW, but with no statistically significant differences found between treatments. Soil ecotoxicity was significantly reduced along with the degradation of atrazine by strain HB-5, and the ecotoxicity of soils with applied fertilizer was below that of treatments without fertilizer. On the seventh day and later, the MN frequencies of all treatments were decreased in the control levels except for AW. Thus, adjusting soil nutrient contents not only promoted strain HB-5 to remove atrazine in soil but also mitigated soil ecotoxicity effects caused by atrazine. These results are important keystones for future remediation of atrazine-contaminated soils.

Genotoxicity evaluation of surface waters located in urban area of Xi’an City using Vicia faba bioassays

In this study, micronucleus (MCN) and chromosome aberration (CA) tests in Vicia faba root tip cells were carried out in order to assess the water quality and the comprehensive genotoxic potential of surface waters located in the urban area of Xi’an City, China. For these evaluations, water samples from different surface waters (four rivers, two lakes, two biological treatment plant effluents) were collected, the ultra-pure water and methyl methanesulfonate solution was used as the negative and positive control, respectively. In our results, highly significant differences in MCN permillage (average number of micronuclei per 1000 cells), CA frequencies and PI (pollution index) values were found among three rivers and two WWTP effluents, the tested samples from two rivers caused the decrease of mitotic index over 22% compared with the negative control. No significant changes were observed in micronuclei and chromosome aberrations frequencies at one river and two lakes during the period of test (wet season). These results point out a poor state of the water quality and genotoxic activity of the main surface waters in Xi’an City. It is recommended to establish a monitoring program for the presence of genotoxic agents in these surface waters.

Genotoxicity of the sediments collected from Pearl River in China and their polycyclic aromatic hydrocarbons (PAHs) and heavy metals

The accelerated industrialization and urbanization in the last three decades around the Pearl River Delta within Guangdong Province in China have led to serious concerns about the impacts on the aquatic environment. In the present study, the genotoxicity of the sediments collected from the Pearl River was evaluated by micronucleus (MN) assay with Vicia faba root tip cells, and the 16 EPA priority polycyclic aromatic hydrocarbons (PAHs) and heavy metals (HMs, including Cr, Cu, As, Se, Cd, Hg, and Pb) in the sediments were determined respectively by GC-MS, inductively coupled plasma mass spectrometry, and inductively coupled plasma atomic emission spectrometry. The results showed that there were significant increases of MN frequencies observed in the sediment-exposed groups, compared with the negative group (P < 0.05, P < 0.01), indicating that the sediments clearly had genotoxicity to the V. faba root cells. The total concentrations of the priority PAHs (250-13,656 ng g(-1), dry weight) and HMs (As, 22,770-36,639 μg kg(-1); Cr, 39,333-133,343 μg kg(-1); Cu, 36,145-159,270 μg kg(-1); Pb, 51,210-166,642 μg kg(-1); Cd, 475.4-1,818.9 μg kg(-1); Hg, 59.9-460.8 μg kg(-1); and Se, 331.7-1,250.4 μg kg(-1), dry weight) were close to those obtained from other urbanized and industrialized areas, which have been considered moderately polluted. There was a clear positive correlation between MN potency and the molar concentrations of Hg and Pb in the sediments (Hg, r = 0.94; Pb, r = 0.91), suggesting that Hg and Pb were the most important factors that posed the sediments higher genotoxicity to V. faba root cells. Our results suggested that both biological and chemical approaches are necessary to be included in a battery of tests to assess the eco-environmental risks of sediments.

Alleviation Effect of Alginate-Derived Oligosaccharides on Vicia faba Root Tip Cells Damaged by Cadmium

Cadmium has been shown to prevent Vicia faba growth by inhibiting cell mitosis. In this study we investigated the role of Alginate-derived Oligosaccharides (ADO) in alleviating Vicia faba root tip cells damaged by 6 and 8 mg L(-1) CdCl2. Micronucleus assay and chromosomal aberration assay were used to determine mitotic index, micronucleus frequency and chromosomal aberration frequency. The results showed that micronucleus frequency of Vicia faba root tip cells was inhibited under all the ADO concentrations. Especially, the inhibition ratio of 0.125% ADO highly reached 66.11 and 67.17% in 6 and 8 mg L(-1) CdCl2, respectively. Furthermore, the mitotic index increased (p < 0.05) and chromosomal aberration frequency decreased (p < 0.05) under all the ADO concentrations. This indicated that ADO had a significant alleviation effect on Vicia faba root tip cells damaged by cadmium.

Induction of sister chromatid exchanges by streptonigrin, an antibiotic and antineoplastic agent

The frequencies of chromosomal aberrations and sister chromatid exchanges, SCE, were analyzed in lateral root-tip cells of Vicia faba treated with the antibiotics streptonigrin or bleomycin. Whereas both agents were very potent inducers of chromosomal aberrations in all phases of the cell cycle, only streptonigrin was able to increase the frequency of SCE. A 3.5-fold increase was obtained with 5 times 10-5 M streptonigrin. A higher yield of chromosomal aberrations is obtained when nitrogen is bubbled through the streptonigrin solution during treatment, than when treatment is carried out in the presence of air. This modification of the treatment conditions, however, does not alter the frequency of SCE. If the results obtained are compared with information in the literature on the types of lesions produced by bleomycin and streptonigrin, it is suggested that neither single- and double-strand breaks, nor apurinic and apyrimidinic sites, give rise to SCE, and that another unknown type of lesion is responsible for the S-dependent action produced by streptonigrin.

Production of sister chromatid exchanges by X-rays under aerobic and anaerobic conditions

Lateral root-tip cells of Vicia faba were irradiated under aerobic and anaerobic conditions with various doses of X-rays and the frequencies of sister chromatid exchanges (SCE) and chromosomal aberrations analyzed. Whereas the frequencies of X-ray-induced chromosomal aberrations were strongly potentiated by oxygen (oxygen enhancement ratio, OER = 2.4), the induction of SCE was comparatively little affected (OER = 1.4). A comparison between these observations and the information available on the influence of oxygen on various types of DNA lesion induced by sparsely ionizing radiation indicates that SCE is likely to be induced by base damage, rather than by strand breaks, whereas both types of lesion might give rise to chromosomal aberrations.

Analysis of SCEs in Vicia faba chromosomes by a simple fluorescent plus Giemsa technique

Cortés, F. and Andersson, H.C. 1987. Analysis of SCEs in Viciafaba chromosomes by a simple fluorescent plus Giemsa technique. —Hereditas 207: 7–13. Lund, Sweden. ISSN 0018–0661. Received October 3, 1986 A modified fluorescent plus Giemsa (FPG) technique for differential staining of sister chromatids and for studying sister chromatid exchanges (SCE) in root tip cells of Viciafaba is described. This new technique is quick and simple and has the advantage of a high reproducibility. The best quality of the sister chromatid differentiation (SCD) is obtained when seedlings are grown for two rounds of chromosome replication in the presence of 5-bromodeoxyuridine (BrdUrd). The technique stains the bifilarly substituted chromatid light and the unifilarly substituted one dark. However. SCD is also obtained after one round of replication in BrdUrd followed by another in the presence of thymidine. There was no difference in the baseline level of SCE in the two types of experimental protocol. The influence of various concentrations of BrdUrd on the yield of SCEs was investigated, as well as the frequency of SCEs in root tip cells grown in the absence and presence of 5-fluorodeoxyuridine. The results of these experiments are discussed.

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.

Leachates of municipal solid waste incineration bottom ash from Macao: Heavy metal concentrations and genotoxicity

Heavy metals in municipal solid waste incineration bottom ash (MSWIBA) may leach into soil and groundwater and pose long-term risks to the environment. In this study, toxicity characteristic leaching procedure (TCLP) was carried out on the MSWIBA from Macao. Heavy metals in leachates were determined by inductively coupled plasma mass spectrometry (ICP-MS) and inductively coupled plasma atomic emission spectrometry (ICP-AES), and genotoxicity of leachates was also evaluated by micronucleus (MN) assay with Vicia faba root tip cells. The results showed that the concentrations of aluminium (Al), manganese (Mn), cobalt (Co), cadmium (Cd) and mercury (Hg) in the leachates were less than 0.01 mg l(-1), and those of iron (Fe), copper (Cu) and molybdenum (Mo) were less than 0.1 mg l(-1). The concentrations of chromium (Cr), zinc (Zn), selemium (Se), strontium (Sr), barium (Ba) and caesium (Cs) were between 0.11 mg l(-1) and 2.19 mg l(-1). Lead (Pb) concentrations, in particular, reached as high as 19.6 mg l(-1), significantly exceeding the maximum concentration limit (5 mg l(-1) for lead by TCLP). Compared with the negative group, a significant increase of MN frequencies was observed in the leachate-exposed groups (P<0.05). With the increase of heavy metals in the leachates, the toxic effects on the Vicia faba root tip cells increased, implying that heavy metals were the main factors causing the genotoxic effects. Our results suggested that apart from chemical analysis, bioassays like the MN assay of Vicia faba root tip cells should also be included in a battery of tests to assess the eco-environmental risks of bottom ashes before decisions can be made on the utilization, treatment or disposal.

Joint effects of microwave and chromium trioxide on root tip cells of Vicia faba

The mutagenic effects of microwave and chromium trioxide (CrO(3)) on Vicia faba root tip were studied. Micronucleus assay and chromosomal aberration assay were used to determine the mitotic index, the micronucleus frequency and chromosomal aberration frequency of Vicia faba root tip cells induced by microwave and CrO(3). The results showed that the micronucleus frequency decreased, and that the mitotic index and chromosomal aberration frequency showed linear dose responses to CrO(3), in treatment of microwave for 5 s. In microwave of 25 s, the mitotic index decreased, the micronucleus frequency and chromosomal aberration frequency increased with increase of CrO(3) concentration. We concluded that microwave and CrO(3) had antagonistic effect on the mitotic index of Vicia faba root tip cells, but had synergetic effect on micronucleus frequency and chromosomal aberration frequency of Vicia faba root tip cells.

Mutagenic effects of chromium trioxide on root tip cells of Vicia faba

In this study on the mutagenic effects of different concentrations of chromium trioxide (CrO(3)) on Vicia faba root tip, micronucleus assay and chromosome aberration assay were used to determine the mitotic indexes, micronucleus aberration rate and chromosome aberration rate of Vicia faba root tip cells. The results showed that the effects of CrO(3) concentration on the mitotic indexes were complicated. CrO(3) increases the micronucleus aberration rate of Vicia faba root tip cells. It was found that within certain range of CrO(3) concentration the micronucleus aberration rate increased systematically with increased concentration of CrO(3), but that the micronucleus aberration rate decreased at higher level of CrO(3) and that CrO(3) also caused various types of chromosome aberration at a rate which increased systematically with increased concentration of CrO(3). We concluded that CrO(3) has significant mutagenic effect on Vicia faba root tip cells.

Inducing somatic meiosis-like reduction at high frequency by caffeine in root-tip cells of Vicia faba

Germinated seeds of Vicia faba were treated in caffeine solutions of different concentration for different durations to establish the inducing system of somatic meiosis-like reduction. The highest frequency of somatic meiosis-like reduction could reach up to 54.0% by treating the root tips in 70 mmol/l caffeine solution for 2 h and restoring for 24 h. Two types of somatic meiosis-like reduction were observed. One was reductional grouping, in which the chromosomes in a cell usually separated into two groups, and the role of spindle fibers did not show. The other type was somatic meiosis, which was analogous to meiosis presenting in gametogenesis, and chromosome paring and chiasmata were visualized.

Bivariate flow cytometry DNA/BrdUrd analysis of plant cell cycle.

We describe a protocol for flow cytometry analysis of cell cycle in plants using indirect immunolabelling staining and Vicia faba, Pisum sativum and Zea mays root tip cells as model systems. The protocol is based on simultaneous analysis of two fluorescent signals. The first, obtained after staining with propidium iodide, is used to quantify nuclear DNA content. The second, obtained after indirect immunofluorescent staining of bromodeoxyuridine (BrdUrd), is used to quantify the amount of BrdUrd incorporated into nuclear DNA. In an attempt to standardize the procedure, the effects of various conditions for partial DNA denaturation using HCl, as well as of BrdUrd concentration and incorporation time on flow cytometry DNA/BrdUrd content analysis have been studied. Maximum BrdUrd-linked fluorescence was observed after a 30 min pulse with 10 microM BrdUrd and after DNA denaturation with 1.5 N HCl (final concentration) for 30 min at 25 degrees C. Under these conditions, DNA content histograms with relatively small coefficient of variation (< 4%, full peak) could be obtained. To avoid non-specific staining of cytoplasm and cell walls, the protocol involves the use of nuclei isolated from formaldehyde-fixed tissues. Fixed isolated nuclei are stable and may be stored in hexylene glycol 0.75 M at 4 degrees C for prolonged periods prior to actual staining and analysis.

Treatment ofVicia fabaroot tip cells with specific inhibitors to cyclin‐dependent kinases leads to abnormal spindle formation

Many events during cell division are triggered by an evolutionary conserved regulator, the cyclin-dependent kinase (Cdk). Here we used two novel drugs, the purine analogues bohemine and roscovitine, to study the role of Cdks in cell cycle progression and microtubule organisation in Vicia faba root tip cells. Both drugs inhibited the activity of immunopurified Vicia faba and alfalfa Cdc2-kinase. The transcript levels of an A- and B-type cyclin, as well as of the cdc2 genes, declined in treated root tips, while the mRNA level of a D-type cyclin gene was not affected. An observed transient arrest at the G1/S and G2/M regulatory points indicated that inhibition of the Cdc2-kinase had an effect on both transitions. In contrast to the regular bipolar spindle in untreated cell, in drug-treated metaphase cells abnormally short and dense kinetochore microtubule fibres were observed. These microtubules were randomly arranged in the vicinity of the kinetochores and connected the chromosomes. Thus, the chromosomes were not aligned on the metaphase plate but were arranged in a circle, with kinetochores pointing inwards and chromosome arms pointing outwards. gamma-Tubulin, which plays a role in microtubule nucleation, also localised to the centre of the monopolar spindle. The observed abnormalities in mitosis, after inhibition of Cdc2-kinase by specific drugs, suggest a role for this enzyme in regulating some of the steps leading to a bipolar spindle structure.

Effects of dNTPs on the SCE frequency in plant root tip cells

The effects of deoxynucleoside triphosphates (dNTPs) on the frequency of sister-chromatid exchange (SCE) have been studied in plant root tip cells. Treatment with dATP caused a dose-dependent decrease in SCE frequency, while dGTP, caused a dose-dependent increase in SCE frequency both in Hordeum vulgare and in Vicia faba root tip cells. Treatment with dCTP and dTTP significantly increased SCE frequency in H. vulgare root tip cells, but significantly decreased SCE frequency in V. faba root tip cells. The SCE induced by treatment with dGTP could be totally reversed by treatment with an equal concentration of any of the other three dNTPs (dATP, dCTP and dTTP) both in H. vulgare and in V. faba. Treatment with a mixed equal concentration of dATP, dCTP, dGTP and dTTP did not alter the SCE frequency. These results suggest that dNTP pool imbalance is an SCE-affecting factor in plant cells.