Investigating the underlying mechanism of cadmium-induced plant adaptive response to genotoxic stress.

Plants as sessile organisms have developed some unique strategies to withstand environmental stress and adaptive response (AR) is one of them. In the present study Cadmium (Cd)-induced AR was evaluated to ameliorate the genotoxicity of a known chemical mutagen ethyl methanesulphonate (EMS) based on cytotoxicity, genotoxicity and oxidative stress in two model plant systems Allium cepa L. and Vicia faba L. Priming the plants with cadmium chloride (CdCl2, 25 and 50 µM) reduced the genotoxicity of EMS (0.25 mM). Cd-induced AR was evident by the magnitude of adaptive response (MAR) values calculated for cytotoxicity, genotoxicity and biochemical parameters. In addition the involvement of some major metabolic pathways and epigenetic modifications in AR was investigated. Metabolic blockers of protein kinase cascades, DNA repair, oxidative stress and de novo translation interfered with the adaptive response implying their role in AR whereas, inhibitors involved in post-replication repair and autophagy were ineffective implicating that they probably have no role in the AR studied. Moreover to find the role of DNA methylation in AR, methylation-sensitive comet assay was carried out. Simultaneously 5-methyl- 2'-deoxycytidine (5mdC) levels were quantified by HPLC (high performance liquid chromatography). AR was eliminated in cells treated with a demethylating agent, 5-aza- 2'deoxycytidine (AZA). Results implied a contribution of DNA hypermethylation. To the best of our knowledge this is a first report correlating DNA methylation to Cd-induced adaptive response in plants undergoing genotoxic stress.

A metabolomic, transcriptomic profiling, and mineral nutrient metabolism study of the phytotoxicity mechanism of uranium.

Uranium (U) is a nonessential element that is readily adsorbed and retained in plant roots, causing root damage plants, rather than being translocated to other parts of the plant. The phytotoxicity mechanism of U is poorly understood. In this study, Vicia faba, a model plant for toxicological research, was selected as experimental material to investigate the phytotoxicity mechanism of U. In this study, the effects of U on the growth and development, methonome, transcriptome and mineral nutrient metabolism of V. faba were studied under different U treatments (0-25 µM) by integrating metabolomics, transcriptomic, and mineral nutrient metabolism analysis techniques. The results showed that U accumulation in roots and aboveground parts reached 164.34-927.90 µg/pot, and 0.028-0.119 µg/pot, respectively. U was mainly accumulated in the cell wall of roots, which damaged the root microstructure and inhibited root growth and development. In terms of mineral nutrient metabolism, U treatment (0-25 µM) led to changes in mineral metabolic profiles of seedlings. In total, 612 different metabolites were identified in nontargeted metabolomics, including 309 significantly upregulated metabolites and 303 significantly downregulated metabolites. Using RNA-seq, 4974 differentially expressed genes (DEGs) were identified under the high-concentration U treatment (25 µM), including 1654 genes significantly upregulated genes and 3320 genes significantly downregulated genes. Metabolic pathway analysis showed that a high concentration of U led to an imbalance of mineral nutrient metabolism in plants and changes in the metabolism and transcriptome pathway of plants, including alterations in the function of plasmodesmata and auxin signal transduction pathway. The latter finding may potentially explain the toxic effect of U on plant roots.

[Influence of Four Kinds of PPCPs on Micronucleus Rate of the Root-Tip Cells of Vicia-faba and Garlic].

In order to determine the degree of biological genetic injury induced by PPCPs, the genotoxic effects of the doxycycline (DOX), ciprofloxacin (CIP), triclocarban (TCC) and carbamazepine (CBZ) in the concentration range of 12.5-100 mg · L⁻¹ were studied using micronucleus rate and micronucleus index of Vicia-fabe and garlic. The results showed that: (1) When the Vicia-faba root- tip cells were exposed to DOX, CIP, TCC and CBZ, micronucleus rates were higher than 1.67 ‰ (CK1), it was significantly different from that of the control group (P < 0.05), and the micronucleus index was even greater than 3.5; With the increasing concentrations of the PPCPs, the micronucleus rates first increased and then decreased. (2) When the garlic root tip cells were exposed to DOX, CIP, TCC and CBZ respectively, the micronucleus rates were less than those of the Vicia-faba, while in most treatments significantly higher than that of the control group (0.67‰). The micronucleus index was higher than 3.5 in the groups exposed to CIP with concentrations of 25, 50, 100 mg · L⁻¹ and TCC and CBZ with concentrations of 25 mg · L⁻¹; With the increase of exposure concentrations, the micronucleus rate showed a trend of first increasing and then decreasing as well. (3) Under the same experimental conditions, the cells micronucleus rates of the garlic cells caused by the four tested compounds were significantly lower than those of Vicia-faba. (4) The micronucleus index of the root tip cells of Vicia-faba and garlic treated with the four kinds of compounds followed the order of CIP > CBZ > TCC > DOX. These results demonstrated that the four compounds caused biological genetic injury to root-tip cells of Vicia-faba and garlic, and the genetic damage caused to garlic was significantly lower than that to Vicia-faba. The damages caused by the four kinds of different compounds were also different.

Molecular and cytogenetic assessment of Dipterygium glaucum genotoxicity.

The aim of the present study is to assess the genotoxicity of Dipterygium glaucum grows widely in Saudi Arabia desert to produce safety herbal products. This work is considered the first and pioneer report so far due to the lack and poor evaluated reports of the plant species for their mutagensity, genotoxicity and cytogenetics effects. Cytogenetic effects of D. glaucum on mitotic in roots of Vicia faba showed reduction in mitotic activity using three extracts; water, ethanol and ethyl acetate. Chromosomal abnormalities were recorded that included stickiness of chromosomes, chromatin bridge, fragments, lagging chromosome and micronuclei. Protein bands and RAPD analyses of V. faba treated with three D. glaucum extracts revealed some newly induced proteins and DNA fragments and other disappeared. Chemical constitution of the plant species should be identified with their biological activities against human and animal cells like HeLa cancer cell line. We are recommending using additional genotoxicity tests and other toxicity tests on animal culture with different concentrations and also utilizing several drought and heat tolerant genes of the plant species in gene cloning to develop and improve other economical crop plants instead of using the species as oral herbal remedy.

Molecular and structural changes induced by essential oils treatments in Vicia faba roots detected by genotoxicity testing.

Over the last few years, there has been an increased interest in exploiting allelopathy in organic agriculture. The aim of this investigation was to examine the effects of essential oil mixtures in order to establish their allelopathic use in agriculture. Two mixtures of essential oils consisting respectively of tea tree oil (TTO) and clove plus rosemary (C + R) oils were tested. Phytotoxicity and genotoxicity tests on the root meristems of Vicia faba minor were performed. A phytotoxic influence was particularly relevant for C + R mixture, while genotoxicity tests revealed significant results with both C + R oil mixture and TTO. Phenotypic analysis on Vicia faba minor primary roots following C + R oil mixture treatment resulted in callose production, an early symptom attributed to lipid peroxidation. The approach described in this study, based on genotoxicity bioassays, might identify specific DNA damage induced by essential oil treatments. These tests may represent a powerful method to evaluate potential adverse effects of different mixtures of essential oils that might be useful in alternative agriculture. Future studies are focusing on the positive synergism of more complex mixtures of essential oils in order to reduce concentrations of potentially toxic components while at the same time maintaining efficacy in antimicrobial and antifungal management.

Liming can decrease legume crop yield and leaf gas exchange by enhancing root to shoot ABA signalling.

To meet future requirements for food production, sustainable intensive agricultural systems need to optimize nutrient availability to maximize yield, traditionally achieved by maintaining soil pH within an optimal range (6-6.5) by applying lime (calcium carbonate). However, a field trial that applied recommended liming rates to a sandy loam soil (increasing soil pH from 5.5 to 6.2) decreased pod yield of field bean (Vicia faba L. cv. Fuego) by ~30%. Subsequent pot trials, with liming that raised soil pH to 6.3-6.7, reduced stomatal conductance (g(s)) by 63, 26, and 59% in V. faba, bean (Phaseolus vulgaris), and pea (Pisum sativum), respectively. Furthermore, liming reduced shoot dry biomass by 16-24% in these species. Ionomic analysis of root xylem sap and leaf tissue revealed a decrease in phosphorus concentration that was correlated with decreased g(s): both reductions were partially reversed by adding superphosphate fertilizer. Further analysis of pea suggests that leaf gas exchange was reduced by a systemic increase (roots, xylem sap, and leaves) in the phytohormone abscisic acid (ABA) in response to lime-induced suboptimal plant phosphorus concentrations. Supplying synthetic ABA via the transpiration stream to detached pea leaves, at the same xylem sap concentrations induced by liming, decreased transpiration. Furthermore, the g(s) of the ABA-deficient mutant pea wilty was unresponsive to liming, apparently confirming that ABA mediates some responses to low phosphorus availability caused by liming. This research provides a detailed mechanistic understanding of the physiological processes by which lime application can limit crop yields, and questions the suitability of current liming recommendations.

Evaluation of an aerobic treatment for olive mill wastewater detoxification.

Olive mill wastewater (OMWW) is a by-product of the olive oil extraction industry. Its dumping creates severe environmental problems in the Mediterranean countries. The phytoxicity of OMWW is due to the phenolic substances and is evaluated through a genotoxicity method. An aerobic treatment of OMWW was conducted during 45 days. Different concentrations of raw and treated OMWW were tested using the Vicia faba micronuclei test. Results showed that raw OMWW induced significant micronuclei formation at 10% of OMWW dilution. At 20% of dilution, no mitosis was recorded. The 45 days aerobic treatment OMWW showed an important decrease in the genotoxicity and also in the toxicity that was observed at 10% and 20% OMWW dilution. This could be correlated with the biodegradation of 76% of the total phenols. Indeed, qualitative analysis by high performance liquid chromatography shows the disappearance of the majority of phenolic compounds after 45 days of treatment. This study was completed by an agricultural test with V. faba plant. Data showed significant growth yield of 36.3% and 29.9% after being irrigated with 5 and 10 t/ha, respectively. These results supported the positive role of aerobic treatment on OMWW and their capacity to ameliorate the agronomic potential of these effluents.

[Allelopathic effects of extracts from fibrous roots of Coptis chinensis on two leguminous species].

An experiment was carried out to study the allelopathic effects of Coptis chinensis fibrous root extracts (CRE) on the germination and seedling growth of Vicia faba and Pisum sativum in order to alleviate the allelopathic effects and increase land productivity. The seeds of both garden pea (P. sativum) and broad been (V. faba) were germinated in CRE solution of various concentrations, the germination rate, seedling growth and related physiological indexes were measured. The result indicated that there were no significant effects of CRE in low concentrations on seed germination, including both the rate and index, and seed vitality and membrane permeability. With the increment of CRE concentrations, however, the high seed membrane permeability and germination inhibition were observed. For example, the germination rates were reduced by 23.4% (P. sativum) and 9.5% (V. faba), respectively, in CRE solution with 800 mg . L-1. Simultaneously, soluble sugars and the free amino acids in the seeds were lower than those in the control (without CRE) after soaking seeds in CRE solutions. In addition, the seedling growth and nitrate reductase activity were stimulated by CRE at low concentrations in contrast to high concentrations which behaved otherwise and inhibited the nutrient utilization in endosperm. Therefore, the large amount of allelochemicals released from the roots and remains of C. chinensis in soils could inhibit the seed germination and seedling growth of legumes, which may lead to decrease even fail crop yields after growing this medical plant.

Determination of protective role of selenium against aflatoxin B1-induced DNA damage.

Selenium is an essential mineral for a healthy life. Appropriate doses of it may undertake a protective role in the organism. In this study, the protective role of selenium (Se(4+)) against aflatoxin B1 (AFB1)-induced DNA damage was determined using random amplified polymorphic DNA on two plants including Vicia faba and Zea mays. It was observed that the concentrations of 0.1, 0.2 and 0.4 ppm of AFB1 have increased polymorphism value, total chlorophyll inhibition rate (IRc, %) and total protein IR (IRp, %). Unlike protein, chlorophyll contents and genomic template stability were decreased. With the addition of different concentrations (0.8 and 80 ppm) of Se(4+) to the treated samples with AFB1, the values return to normal. An 800-ppm concentration of Se(4+), on the contrary, could not inhibit the toxicity of AFB1 but caused an increase in toxicity level of AFB1/enhanced the toxicity level of AFB1. Results suggested that Se(4+) has an antagonistic effect against AFB1 toxicity and that the degree of antagonistic effect of Se(4+) against AFB1 was related to its concentration.

Sucrose synthase activity and carbohydrates content in relation to phosphorylation status of Vicia faba root meristems during reactivation from sugar depletion.

Carbohydrate starvation of Vicia faba root meristems leads to readjustment of carbohydrate metabolism and blocks the cell cycle in two principal control points (PCP1/2). The cell cycle reactivation is possible after sucrose provision, although with a delay of about 12h. During this period, the cells are sensitive to 6-dimethylaminopurine (6-DMAP) and okadaic acid (OA), inhibitors of protein kinases and phosphatases, respectively. The aim of the present study was to investigate whether those inhibitors are involved in inhibition of cell cycle revival through interference with the activities of two sucrose-cleaving enzymes: sucrose synthase (SuSy; EC 2.4.1.13) and invertase (INV; EC 3.2.1.26). In sugar-starved cells, the in situ activity of both enzymes decreased significantly. Following supplementation of root meristems with sugar, INV remained inactive, but SuSy activity increased. Despite the lack of INV activity, glucose was present in meristem cells, but its content was low in cells treated with OA. In the latter case, the size of plastids was reduced, they had less starch, and Golgi structures were affected. In sugar-starved cells, SuSy activity was induced more by exogenous sucrose than by glucose. The sucrose-induced activity was strongly inhibited by OA (less by 6-DMAP) at early stages of regeneration, but not at the stages preceding DNA replication or mitotic activities. The results indicate that prolongation of regeneration and a marked decrease in the number of cells resuming proliferation (observed in previous studies) and resulting from the action of inhibitors, are correlated with the process of SuSy activation at the beginning of regeneration from sugar starvation.

Antioxidant and prooxidant effects of lanthanum ions on Vicia faba L. seedlings under cadmium stress, suggesting ecological risk.

The present study combined chemical analyses and biological measurements to investigate biphasic effects of La on Cd stress in leaves of Vicia faba seedlings, which were hydroponically cultivated for 15 d in the combination of 6 µM CdCl(2) and 2 to 480 µM La(NO(3))(3), respectively. The results showed that contents of Cd first elevated above and then declined below the 6 µM single Cd treatment when 2 to 30 µM extraneous La were combined. Contents of mineral nutrients altered differentially and became imbalanced. No distinct band was observed in catalase (CAT), guaiacol peroxidase (GPX), or ascorbate peroxidase (APX) patterns, but in superoxide dismutase (SOD) isozymes by the supplementation with 8 to 480 µM of extraneous La. Superoxide dismutase and APX activities changed as a U-shaped curve; however, CAT and GPX changed as an inverted U-shaped curve along with increasing La. Moreover, heat shock protein 70 (HSP 70) production was reduced below the single treatment of Cd at 2 to 8 µM of extraneous La and enhanced thereafter. Thus, La at lower concentrations promoted antioxidation against Cd stress; La at higher concentrations turned to prooxidant effects, implicating potential ecological risk. Heat shock protein 70, combined with the antioxidant enzymes, constitutes an integrative defense system, which can be used to estimate the degree of antioxidation or prooxidation of extraneous La to Cd-induced oxidative stress in the seedlings.

[Genetic effects of root extracts of Glycyrrhiza glabra L. on different test-systems].

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

The influence of selenium on root growth and oxidative stress induced by lead in Vicia faba L. minor plants.

The effect of selenium (Se) on Vicia faba L. minor roots subjected to lead (Pb) stress was studied by investigating root growth, root viability, and antioxidant enzyme activity. The experiments were carried out on plants grown for 2 weeks on Hoagland medium supplied with 50 µM Pb in the form of lead nitrate Pb(NO(3))(2) and/or Se concentrations of 1.5 and 6 µM in the form of sodium selenite Na(2)SeO(3). It was shown that Pb reduced the root growth and caused serious damage in the roots, which was accompanied by metal accumulation in these tissues. The exposition of roots to Pb led to significant changes in the biochemical parameters: the MDA and T-SH content and glutathione peroxidase (GSH-Px) activity increased but the guaiacol peroxidase (GPOX) activity decreased. Moreover, Pb intensified O(2)(·-) production in the roots. Selenium at a lower concentration alleviated Pb toxicity which was accompanied by a decreased O(2)(·-) production in the apical parts of roots and increased the T-SH content and GPOX activity. However, higher Se concentration intensified MDA and T-SH accumulation and GPOX and GSH-Px activity in Pb-treated plant roots. At low concentration, Se improved cell viability whereas at high concentration it was pro-oxidant and enhanced the lipid peroxidation and cell membrane injury.

Biphasic effects of lanthanum on Vicia faba L. seedlings under cadmium stress, implicating finite antioxidation and potential ecological risk.

In the present study, lanthanum (La) as a representative REE was used to explore the mechanisms for alleviation of Cd-induced oxidative damage by extraneous La at appropriate concentrations, and to assess ecological risk of combination of Cd and La at higher concentrations in roots of Vicia faba L. seedlings. The seedlings were hydroponically cultured for 15 d under nutrient solution, 6 µmol L(-1) CdCl(2), and combination of 6 µmol L(-1) CdCl(2) and increasing concentrations of La, respectively. The results showed that the supplementation with low concentrations of exogenous La (<120 µmol L(-1)) led to reduced contents of Cd, Ca, Cu, Zn, Mn or Fe element and increased activities of superoxide dismutase (SOD), catalase (CAT), guaiacol peroxidase (GPX) and ascorbate peroxidase (APX) isozymes as well as heat shock protein 70 (HSP 70) production in the roots. However, the supplementation with higher La (>120 µmol L(-1)) showed the adverse effects. The contents of Cd elevated above the single Cd treatment in the roots, accompanying with the decline of antioxidant isozyme's activities and HSP 70, and increment of carbonylated proteins and endoprotease isozyme's activities. The results also showed that the root growth was not only related to carbonylated proteins, but also to indole acetic acid oxidase activities. Therefore, the supplemented extraneous La contributed to biphasic effects: stimulated antioxidation at lower concentrations and pro-oxidation at higher concentrations against Cd-induced oxidative stress in the roots.

[Genoprotective activities of the oils from leaves and fruits of Fagus orientalis Lipsky].

The antimutagenic activities of the oils obtained from leaves and fruits of Fagus orientalis have been shown in experiments with spontaneous and mutagen- and ageing-induced variability. The aberrations of chromosomes in the meristematic cells of the Allium cepa L., Vicia faba L., Triticum aestivum L., and marrow cells of Vistar rats as well as Arabidopsis thaliana gene mutations have been mobilized as experimental tests.

Lanthanum resulted in unbalance of nutrient elements and disturbance of cell proliferation cycles in V. faba L. seedlings.

Effects of lanthanum (La) on mineral nutrients, cell cycles, and root lengthening have been little reported. The present work investigated these physiological responses in roots of Vicia faba seedlings cultivated in La3+-contained solutions for 15 days. The results showed that the increasing contents of La in the roots and leaves contributed to disbalances of contents of Ca, Fe, Cu, Zn, Mg, Mn, P, and K elements, and potential redistributions of some elements in the roots and leaves. These disbalances might be involved in the subsequent alteration of cell cycle phases in the root tips. Low-dose promotion and high-dose inhibition (Hormetic effects) were demonstrated as the dose responses of G0/G1-, S- or G2/M-phase ratios. The cell cycles were most probably arrested at G1/S interphase by La3+ in the root tips. The fact that the root lengths were not consistent with the changes of cell cycle phases suggested that the cell proliferation activities might be masked by other factors (e.g., cell expansion) under long-time exposure to La3+.

Protective role of Ginkgo biloba on petroleum wastewater-induced toxicity in Vicia faba L. (Fabaceae) root tip cells.

The Melet is one of Turkey's economically important rivers. Most of the petroleum plants are located at fairly nearby of the river This situation is considered as main source of heavy metal pollution in the river. The present study was designed to evaluate the protective role of Ginkgo biloba (GB) on cytotoxicity induced by petroleum wastewater in Vicia faba root tip cells. For this aim, we used the germination percentage, root length, weight gain and micronucleus (MN) frequency as indicators of cytotoxicity. Additionally to the cytological analysis, lipid peroxidation analyses were also performed in V. faba roots. Heavy metal concentrations in wastewater were measured by atomic absorption spectrophotometer (AAS). The V. faba seeds were divided into six groups. They were treated with petroleum wastewater and 10, 20 and 30 microM doses of GB. As a result, the mean concentrations of heavy metals in wastewater were observed in the order: Pb > A > Ni > Cr > Fe > Cu > Zn > Cd. The highest germination percentage was observed in the seeds of the control and positive control groups (in proportion as 98 and 96%, respectively). Wastewater treatment caused a significant decrease in the germination percentage of Group III (in proportion as 44%). The highest root length and weight gain were observed in the seeds of the control and positive control groups at the end of the experimental period. The least root length and weight gain were observed in the seeds of Group III treated with wastewater alone. In the control group, the final weights of all the seeds increased about 4.08 g according to initial weight. The root lengths of the control seeds were measured as 6.80 cm at the end of the experimental period. The final weights of the seeds exposed to wastewater alone increased about 0.90 g according to initial weight. Besides, there was a significantly increase in the MDA levels of the roots exposed to wastewater. Heavy metals in wastewater significantly affected the MDA production indicating lipid peroxidation. But, GB-treatment caused amelioration in indices of the germination percentage, root length, weight gain, MN frequency and lipid peroxidation when compared with group III. Each dose of GB provided protection against wastewater toxicity and its strongest protective effect observed at dose of 30 microM. In vivo results showed that GB is a potential protector against toxicity induced by petroleum wastewater and its protective role is dose-dependent.

Efficiency of propolis extract on faba bean plants and its role against nematode infection.

Propolis is a resinous substances collected by honey bees, It cannot be used as a raw material, so it must be purified by extraction with solvent. This work aimed to study the physiological influence of propolis extract as foliar application or soil drench on faba bean plants and its role against nematode infection. Propolis samples were extracted by using three different solvents (distilled water or 70% ethanol or acetone) (750 and 1000 mg/l). Qualitative tests of the propolis extracts proved that these extracts contain sterols, flavonoids and phenolic compounds. Moreover, few numbers of phenolic acids (coumaric, ferulic, salicylic and benzoic acid) were also detected on TLC plates. All treatments of propolis extract (as foliar application or soil drench) increased total chlorophyll and carotenoid and the magnitude of increase was more pronounced by applying the higher concentration (1000 mg/l). Moreover, alll propolis extracts increased shoot height; root dry weight; number of branches and pods/plant; number of seeds/pod as well as seed index. Acetonic extract was the most effective particularly at higher concentration. Applying propolis extracts as foliar application or soil drench caused an increase in carbohydrate content of the yielded seeds accompanied by a decrease in phytic acid and vicine content. In addition, foliar application of all propolis extracts caused an increasing in protein content and phenolic compounds of the yielded seeds, whereas, soil drench with ethanolic or acetonic extract only at 1000 mg/L increased protein content. The data revealed that the propolis extract as soil drench reduced the juvenile-Meloidogyne sp.-population density per one kg soil and number of root-galls per one gm roots specially at the higher concentration (1000 mg/l). It is worthy to mention that faba bean plants treated with propolis extract either as foliar application or soil drench could overcome the inhibitory influence of nematode infection.

Use of plant genotoxicity bioassay for the evaluation of efficiency of algal biofilters in bioremediation of toxic industrial effluent.

The toxicity and efficacy of an algal-based bioremediation technology were assessed through bioassays for ecological risk of contaminated industrial effluents. The algal bioremoval of heavy metals was evaluated using an in vitro approach. Phytogenotoxicity tests were conducted with Allium cepa and Vicia faba plants to evaluate the genotoxicity of the industrial effluents before and after treatment with different kinds of algal biofilters (BF). Root cells were exposed for 24 h to different dilutions of both raw and treated effluent of a chemical fertilizer factory. Three cytogenetic endpoints were used to assess the mutagenic potencies of the industrial effluent: mitotic inhibition, mitotic chromosome aberrations, and nuclear irregularities in interphase cells. Before algal treatment, the industrial effluent caused strong genotoxic effects represented by severe inhibition in mitotic activity of meristematic cells and high frequency of both chromosome and nucleus abnormalities. After algal treatment, the cytotoxic effects of 30% and 60% concentrations of the treated effluent were comparable to those of 5% and 10% concentrations before treatment, respectively, and the frequency of both chromosome and nuclear abnormalities declined by approximately 50%. Statistical analysis of the data indicates a significant reduction in genotoxicity associated with a remarkable reduction in heavy metal concentrations after bioremediation by algal BF. The Allium and Vicia genotoxicity approach was effective in monitoring bioremediated effluent for toxicity.

Phosphorylation of H2AX histones in response to double-strand breaks and induction of premature chromatin condensation in hydroxyurea-treated root meristem cells of Raphanus sativus, Vicia faba, and Allium porrum.

Histone H2A variant H2AX is rapidly phosphorylated on the induction of DNA double-strand breaks by ionizing radiation and hydroxyurea-mediated replication arrest, resulting in the formation of gamma-H2AX foci along megabase chromatin domains nearby the sites of incurred DNA damage. In an attempt to establish a relationship between species-specific nuclear architecture and H2AX phosphorylation in S/G(2) phase-arrested root meristem cells, immunocytochemical comparisons using an antibody raised against human gamma-H2AX were made among three plants differing with respect to DNA contents: Allium porrum, representing a reticulate type of DNA package, Vicia faba, having semireticulate cell nuclei, and Raphanus sativus, characterised by a chromocentric type of chromatin. Another approach was aimed at determining possible correlations between the extent of hydroxyurea-induced phosphorylation of H2AX histones and the quantities of root meristem cells induced by caffeine to enter aberrant mitotic division (premature chromosome condensation). It was concluded that the higher-order structure of chromatin may contribute to the accessibility of molecular factors engaged in the recognition and repair of genetic lesions. Consequently, in contrast to A. porrum and V. faba, a diffuse chromatin in chromocentric cell nuclei of R. sativus may become more vulnerable both to generate DNA double-strand breaks and to recruit molecular elements needed to arrange the cell cycle checkpoint functions, and thus, more resistant to factors which allow the cells to enter premature chromosome condensation spontaneously. On the other hand, however, caffeine-mediated overriding of the S-M checkpoint control system resulted in the typical appearance of premature chromosome condensation, irrespective of the genomic content of DNA.