Faba Root Research Articles

Effect of the Energy Drink "Bison" on Mitosis And Nucleic Acids Content In Vicia faba

The present investigation had been conducted to study the mutagenic effects of four concentrations of energy drink (50, 80, 90, and 100%) with three exposure times (6, 12, and 24hours)on root meristems of Vicia faba plant. Mitotic index was reduced in all treatments except for two treatments: 90% Bison with two exposure times 6 and 12 h. On the other hand, total abnormalities were highly increased in all treatments. The most common abnormalities were: Stickiness, disturbed, C-metaphase, star shape and lagging chromosomes. In addition, bridges, multipolar, breaks, (micro-multi) nuclii and dinucleated occurred with very low frequencies in some treatments. These results-indicated that Bison had Turbagenic effects. Unbalanced mitotic stages percentages occurred as a result of all treatments compaired with control. Different Bison treatments caused altering of DNA and RNA contents in Vicia faba root tips. The inhibitory effects of most Bison treatments were found to depend on exposure time dose. Hundred percentage Bison treatment for 24h caused the highest total abnormalities (90.32%) and the lowest content of DNA and RNA (4.63 and 1.98 mg\g) respectively compared with respective control 2.12%,9.73 and 3.26 mg\g respectively.These findings indicate the mutagenic potentiality of the energy drink Bison.

Sequential effects of cadmium on genotoxicity and lipoperoxidation in Vicia faba roots

Kinetics of stress responses to Cd exposure (50, 100 and 200 μM) expanding from 12 to 48 h were studied in roots of hydroponically cultivated-Vicia faba seedlings. The heavy metal induced toxicity symptoms and growth arrest of Vicia roots gradually to the Cd concentration and duration of the treatment. The intracellular oxidative stress was evaluated with the H(2)O(2) production. The H(2)O(2) content increased gradually with the sequestered Cd and root growth inhibition. Lipid peroxidation-evidenced by malondialdehyde (MDA) content and Evans blue uptake-and genotoxicity-evidenced by mitotic index (MI) and micronuclei (MCN) values-were concomitantly investigated in root tips. By 12 h, root meristematic cells lost 15% of their mitotic activity under 50 or 100 μM Cd treatment and 50% under 200 μM Cd treatment and led cells with MCN, while the MDA content and Evans blue absorption were not affected. The loss of membrane integrity occurred subsequently by 24 h. The increase in MDA content in root cells treated with 50, 100 and 200 μM Cd was significantly higher than the control. By 48 h, the MDA content increased 134, 178 or 208% in root cells treated with 50, 100 and 200 μM Cd, respectively. The Evans blue absorption was also affected by 24 h in roots when treated with 200 μM Cd and gradually increase by 48 h with the Cd concentration of the treatment. The decrease of mitotic activity triggered by 12 h was even higher by 24 h and the MI reduced to 44, 56 or 80% compared to the control in the three different Cd concentrations tested. The different kinetics of early in vivo physiological and cytogenetic responses to Cd might be relevant to the characterization of its toxicity mechanisms in disrupting primarily the mitosis process.

Lead-induced genotoxicity to Vicia faba L. roots in relation with metal cell uptake and initial speciation

Formation of organometallic complexes in soil solution strongly influence metals phytoavailability. However, only few studies deal with the influence of metal speciation both on plant uptake and genotoxicity. In the present study, Vicia faba seedlings were exposed for 6 h in controlled hydroponic conditions to 5 μM of lead nitrate alone and chelated to varying degrees by different organic ligands. Ethylenediaminetetraacetic acid and citric acid were, respectively, chosen as models of humic substances and low weight organic acids present in natural soil solutions. Visual Minteq software was used to estimate free lead cations concentration and ultimately to design the experimental layout. For all experimental conditions, both micronucleus test and measure of lead uptake by plants were finally performed. Chelation of Pb by EDTA, a strong chelator, dose-dependently increased the uptake in V. faba roots while its genotoxicity was significantly reduced, suggesting a protective role of EDTA. A weak correlation was observed between total lead concentration absorbed by roots and genotoxicity ( r 2=0.65). In contrast, a strong relationship ( r 2=0.93) exists between Pb 2+ concentration in exposure media and genotoxicity in the experiment performed with EDTA. Citric acid induced labile organometallic complexes did not demonstrate any significant changes in lead genotoxicity or uptake. These results demonstrate that metal speciation knowledge could improve the interpretation of V. faba genotoxicity test performed to test soil quality.

Study on adverse impact of e-waste disassembly on surface sediment in East China by chemical analysis and bioassays

The primitive recycling of electronic waste (e-waste) in developing countries is causing serious environmental pollution. The objective of this study was to determine the contamination and toxicity of surface sediment of Nanguan River, which runs through the e-waste recycling area of Taizhou, East China. Surface sediments were collected from Nanguan River, including one from the control site, four near the household workshops, and two near the industrial parks. Levels of polychlorinated biphenyls (PCBs), polycyclic aromatic hydrocarbons (PAHs), and heavy metals/metalloid (Ni, Pb, Cd, Cr, Zn, Cu, Hg, and As), which are most widely detected in e-waste contaminated surroundings, were determined. Acute toxicity and genetic toxicity were evaluated by luminescence inhibition assay in luminescent bacterium Vibrio qingaiensis sp. nov. (Q67) and Vicia faba roots tip micronucleus assay, respectively. The surface sediment has been seriously contaminated by PCBs, PAHs, and heavy metals/metalloid due to the e-waste disassembly process. Significant acute and genetic toxicity of the sediments presented a big threat to the aquatic life and human health through food chain, as the river is an extent water source for local residents. The environmental issue of e-waste recycling is emergent and measures should be taken to mitigate the adverse impacts of e-waste disassembly.

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.

New direct contact approach to evaluate soil genotoxicity using the Vicia faba micronucleus test

A method to assess micronucleus (MN) induction in Vicia faba roots by direct contact exposure to a solid matrix was developed. The procedure comprised a 5-d germination period, as in the well-known method using aqueous extracts. However, the seeds were here sown directly into the test soil whereas a culture period is necessary before exposing seedlings to a liquid medium. One soil under forest and two contaminated soils from areas affected by industrial installations and a coke works were used. Three durations of direct exposure were tested: 2, 5 and 7 d. The optimal duration was evaluated at 2 d to observe maximal MN induction without observing toxicity symptoms. The methodology using aqueous extracts was applied to the same three soils: MN frequency was higher than in the direct contact assay but the ratios of MN frequencies from tested soils in comparison to the negative control were lower. However, for each soil, both the direct contact method and the aqueous extract exposure led to the same risk assessment diagnosis. The evaluation of a concentration range of a polycyclic aromatic hydrocarbons (PAH)-contaminated soil showed a dose-dependent MN frequency when the seeds were allowed to germinate before sowing in the soil: the soil genotoxicity was the highest at intermediate doses. The direct contact method was found to be rapid, sensitive and well suited to the evaluation of soil quality.

Use of Antimicrobial and Genotoxicity Potentiality for Evaluation of Essential Oils as Food Preservatives

In this study, investigations were carried out to assess the efficiency of two plant essential oils; rosemary and oregano as natural food preservatives. The effect of the plant essential oils at concentrations of 0.1%, 0.5% and 1% was studied in the soft cheese against Salmonella enteritidis and Escherichia coli at fridge temprature over a 14-day period. The essential oils performed well in the inhibition of S. enteritidis and E. coli. It is concluded that selected plant essential oils can act as potent inhibitors of both microorganisms in a food product. At the same time, evaluation of their safety as food preservatives was undertaken via monitoring the genotoxic activity of the mentioned essential oils using Vicia faba test. Vicia faba roots were treated with the above concentrations for 3 hours. Results revealed cytotoxic and genotoxic effects of the applied doses. Mitotic index decreased significantly when compared to control. Chromosomal abnormalities and micronuclei were also observed and the effects were dose-dependent. Despite the efficient role of the studied oils as antimicrobial agents, their genotoxicity potential in eukaryotic cells made them unacceptable as food preservatives, particularly at high doses. Therefore, more research in the use of essential oils as food preservatives is needed.

Effects of atrazine on dna damage and antioxidative enzymes in Vicia faba

To evaluate atrazine (2-chloro-4-ethylamino-6-isopropylamino-1,3,5-triazine) ecotoxicology in soil, the effect of atrazine on the activity of antioxidative enzymes (superoxide dismutase [SOD], catalase [CAT], and guaiacol peroxidase [POD]) was investigated in Vicia faba roots. Tissues from each treatment were collected on the days 7, 14, 21, and 28. Compared with the controls, SOD activity in V. faba roots was stimulated by the 2.5 mg/kg treatment and inhibited by the 5 and 10 mg/kg treatments, and CAT and POD activities in the 10 mg/kg treatment were inhibited on the whole. The Olive tail moments of single-cell gel electrophoresis of root cells were enhanced after treatment with different doses of atrazine on days 7, 14, 21, and 28, and significant differences were found compared to the controls. In conclusion, atrazine induces oxidative stress and DNA damage on V. faba.

In situ activities of hexokinase and fructokinase in relation to phosphorylation status of root meristem cells of Vicia faba during reactivation from sugar starvation

The plant cell cycle is equipped with two principal control points: PCP1 in G1 and PCP2 in G2 phase. These checkpoints can arrest the cell cycle in response to carbohydrate starvation, while sugar presence can revive the replication and mitotic activity. The process of cell cycle revival is strongly repressed by okadaic acid (OA) or 6-dimethylaminopurine (6-DMAP), inhibitors of specific protein phosphatases 1 or 2A or kinases (cyclin-dependent kinases), respectively. In the present study, it was investigated whether inhibition of cell cycle revival is performed through interference of the above-mentioned inhibitors with the metabolic pathway of sucrose applied to the cells. Changes of hexokinase (HK) and fructokinase (FK) activities, key enzymes of hexose metabolism, were analyzed in Vicia faba root meristem cells arrested in G1 and G2 phase by carbohydrate starvation as well as in those recovered with glucose or sucrose in the presence of OA or 6-DMAP. It was shown that in the sugar-starved cells, the activity of both enzymes decreased significantly. During cell regeneration with carbohydrates, the activity of HK was induced more by sucrose than by glucose, while FK remained inactive after glucose addition. Moreover, in situ investigation of the activities of HK and FK showed that OA-induced and 6-DMAP-induced repression of the cell cycle revival is connected with the interference of these drugs in the metabolic pathway of sucrose. It was also indicated that stronger OA-induced and 6-DMAP-induced inhibition of the replication and mitosis revival, at the early stages of sucrose regeneration, was correlated with the stronger influence of these inhibitors on HK and FK activities.

Aluminum induces chromosome aberrations, micronuclei, and cell cycle dysfunction in root cells of Vicia faba

Aluminum (Al) exists naturally in air, water, and soil, and also in our diet. Al can be absorbed into the human body and accumulates in different tissues, which has been linked to the occurrence of Alzheimer's disease and various neurological disorders. By using Vicia cytogenetic tests, which are commonly used to monitor the genotoxicity of environmental pollutants, cytogenetic effects of aluminum (AlCl(3)) were investigated in this study. Present results showed that Al caused significant increases in the frequencies of micronuclei (MN) and anaphase chromosome aberrations in Vicia faba root tips exposed to Al over a concentration-tested range of 0.01-10 mM for 12 h. The frequency of micronucleated cells was higher in Al-treated groups at pH 4.5 than that at pH 5.8. Similarly, AlCl(3) treatment caused a decrease in the number of mitotic cells in a dose- and pH-dependent manner. The number of cells in each mitotic phase changed in Al-treated samples. Mitotic indices (MI) decreased with the increases of pycnotic cells. Our results demonstrate that aluminum chloride is a clear clastogenic/genotoxic and cytotoxic agent in Vicia root cells. The V. faba cytogenetic test could be used for the genotoxicity monitoring of aluminum water contamination.

Comparison Between New Wild Plant Trifolium repens and Vicia faba on their Sensitivity in Detecting the Genotoxic Potential of Heavy Metal Solutions and Heavy Metal-Contaminated Soils

The DNA damage and cytotoxicity induced by CdCl2 solutions and soils anthropogenically contaminated with heavy metals were studied using the micronucleus (MN) test. Vicia faba, a plant model widely used in liquid exposure assays, was adapted for direct exposure to a solid phase. In addition, the MN assay was adapted to a new wild plant system, the white clover (Trifolium repens). The results obtained after exposure to CdCl2 solutions confirmed that V. faba root cells were a sensitive model and revealed that T. repens root cells were not appropriate for the detection of micronuclei (although they were highly sensitive to the cytotoxic effect of CdCl2). Concerning the results observed after direct exposure to contaminated soils (solid-phase exposure), the MN frequency scores in V. faba root cells were increased in a statistically significant and dose-related manner compared to the control plants. Regarding T. repens root cells, this solid phase exposure confirmed that this model is not appropriate for use in the micronucleus assay.

Enhancement by methylated oxypurines of the frequency of induced chromosomal aberrations.

Eight methylated oxypurines were tested for their ability to potentiate the chromosome damage produced by thiotepa and maleic hydrazide in bean (Vicia faba) roots and by thiotepa in Chinese hamster cells. Caffeine was the strongest potentiating agent in Vicia faba roots, 8-chlorocaffeine in Chinese hamster cells. In both materials tetramethyluric acid was the least active of the methylated oxypurines tested. Caffeine, theobromine and theophylline were effective potentiating agents but poor inducers of exchange-type aberrations in prophase and the post-synthetic period of interphase. Tetramethyluric acid, in contrast, was a poor potentiating agent but effective inducer of exchange-type aberrations. In Vicia faba, post-treatments with caffeine as potentiating agent reduced by a factor of about 4.5 the concentration of thiotepa required to produce a given aberration frequency.

The essential oils in agriculture as an alternative strategy to herbicides: a case study

In this study we investigated the allelopathic effect of some essential oils extracted from aromatic plants to inhibit weed seed germination. The study was performed to evaluate the potential genotoxic effect of lavender on Vicia faba root meristems by genotoxicity tests (comet assay and micronuclei test) and to evaluate microbial diversity fluctuation by Restriction Fragment Length Polymorphism (RFLP) molecular analysis on ecological, biological and conventional soil. The results showed a phytotoxicity effect of the essential oil for all the concentration tested, while there is not a dose-dependent relation between oil concentrations and the genotoxic effect. A fluctuation of microbial communities in relation to the release of essential oils by Lavandula officinalis was also observed. Then we suggest that the essential oils could be useful as potential bioherbicides as an alternative strategy to the chemical remedy, but further studies are necessary to evaluate their appropriate use in the field.

Protein phosphorylation in Vicia faba root meristem cells during the first steps of leaving principal control points after sucrose application

Before Vicia faba root meristem cells stopped by carbohydrate starvation in principal control points (PCP1 and PCP2) start sucrose induced replication and division they go through a phase of metabolic regeneration. This interval is characterised st great sensitivity to the inhibitors of cyclin-dependent protein kinases and protein phosphatases (PPs). In the present research, changes of phosphoprotein levels in the nucleolus, nucleus and cytoplasm were analysed using okadaic acid and 6-dimethylaminopurine (6-DMAP) during the first period of cell regeneration in sucrose (0-3 h). It was established that when the cells start to leave checkpoints, the balance between protein phosphorylation and dephosphorylation shifts towards the intensified activity of PPs. Furthermore, it was also established that the structures appearing during cell regeneration, which were located around cell nuclei and which contained large amounts of phosphorylated proteins, were plastids. The reactions of protein phosphorylation which took place in the plastids were directly correlated with starch synthesis and were stopped by inactivation of protein phosphatases (PP1 and/or PP2A).

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.

Exposure of Vicia faba and Pisum sativum to copper-induced genotoxicity.

The potential genotoxicity of Cu(2+) was investigated in Vicia faba and Pisum sativum seedlings in hydroponic culture conditions. Cu(2+) caused a dose-dependent increase in micronuclei frequencies in both plant models. Cytological analysis of root tips cells showed clastogenic and aneugenic effects of this heavy metal on V. faba root meristems. Cu(2+) induced chromosomal alterations at the lowest concentration used (2.5 mM) when incubated for 42 h, indicating the potent mutagenic effect of this ion. A spectrum of chromosomal abnormalities was observed in V. faba root meristems, illustrating the genotoxic events leading to micronuclei formation.

Environmental, genetic and cellular toxicity of tenuazonic acid isolated from Alternaira alternata

Alternaria alternata, an important pathogen of many plants, produces tenuazonic acid (TeA) with bioactivity to microbes, plants and animals. TeA is one of the main mycotoxin to humans and other organisms. Using Chlamydomonas reinhardtii, Vicia faba root tip and three mammalian normal cell lines as target materials, environmental toxicity, genetic toxicity and cytotoxicity of TeA were examined. The growth and chlorophyll concentration of C. reinhardtii were inhibited at above 100
g/ml concentration, with EC50(G) of 310.36
g/ml and EC50(Chl) of 294.27
g/ml. The micronuleus test results indicated that MCN‰ was >15‰ only at higher concentrations. TeA inhibited the proliferation and total protein contents of 3T3 mouse fibroblasts (3T3 cells), Chinese hamster lung cells (CHL cells) and human hepatocytes (L-O2 cells) at concentrations ranging from 12.5 - 400
g/ml. Of the three cell lines, 3T3 cells were the most sensitive to the toxin (EC50(24h) = 41.64
g/ml), followed by CHL cells (EC50(24h) = 59.33
g/ml), and L-O2 cells (EC50(24h) = 85.98
g/ml) had the lowest sensitivity.

Red cabbage extract limits copper stress injury in meristematic cells of Vicia faba

Natural phenolic compounds (phenolic acids, flavonoids, tannins, lignans) present in food of plant origin are in the focus of interest due to their prevalence, properties and biological activity. The aim of the presented work was to investigate antioxidant and antigenotoxic effects of the anthocyanin-rich extract from red cabbage leaves (Brassica oleracea rubrum) on the changes induced by toxic Cu2+ concentrations. MeOH extract from red cabbage containing anthocyanin (ATH) and phenolic acid derivatives exhibited strong antioxidant properties. Cu2+ decreased mitotic index (MI) and inhibited proliferative activity of Vicia faba root meristematic cells. The morphology of mitotic chromosomes was changed; “erosion” and pulverization might result from Cu2+ high-cytotoxicity. Numerous micronuclei, chromatid bridges and lagging/lost chromosomes were found in the meristematic cells of V. faba, which indicate the clastogenic effect of Cu2+. The application of the ATH-rich extract lowered the number of disturbances induced by Cu2+. The positive role of the ATH-rich red cabbage extract will be 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.

An improved method for the cell cycle synchronization of Vicia faba root meristem cells

Plant root meristem cells divide asynchronously which makes biochemical analysis of cell cycle regulation particularly difficult. In the present article a high level of cell cycle synchronization in Vicia faba root meristems was obtained by using a rich medium (HNS), special culture conditions and a double-block method with replication inhibitor—hydroxyurea (HU). Two HU concentrations were tested and different periods of the first and the second synchronization, and of cycle recommencement between the first and the second blockage. The level of synchronization was estimated on the basis of 3H-thymidine labeling indices, mitotic, and phase indices and indices determining the percentage of G1 and G2 cells, which were identified by cytophotometric measurements of DNA content in individual nuclei. The highest level of cell cycle synchronization was obtained after double treatment of meristems with 1.25 mM HU (18 and 12 h) separated by 6-h incubation in HNS without HU. During the second postincubation in HNS in subsequent hours: 4, 7, 10, 11, over 90% of cells in the S phase, nearly 70% in G2 phase, 86% in mitosis, and nearly 70% in G1 phase were received, respectively. The use of 2.5 mM HU in a similar experimental procedure caused disturbed divisions.