Evaluation of cytological effects of Zn2+ in relation to germination and root growth of Nigella sativa L. and Triticum aestivum L

The effects of different treatments with zinc sulfate (Zn(2+)) on the cytology and growth of Nigella sativa and Triticum aestivum were investigated. Five concentrations of zinc sulfate ranging from 5 to 25mg/l were applied for 6, 12, 18, and 24h. The treatments reduced the germination percentages of N. sativa seeds and T. aestivum grains and inhibited the root growth of both plants. Concentrations higher than 25mg/l of Zn(2+) applied for 24h were toxic for both plants. The non-lethal concentrations of Zn(2+) showed an inhibitory effect on cell division in root tips of both plants and caused a decrease in their mitotic index values. The reduction in MI in root tips of T. aestivum was more evident than that of N. sativa. All treatments changed the frequency of mitotic phases as compared with the control values. The total percentage of abnormalities in N. saliva was more than that in T. aestivum. Zn(2+) treatments produced a number of mitotic abnormalities in dividing cells in root tips of both plants resulting from its action on the spindle apparatus such as C-metaphases, lagging chromosomes and multipolar anaphases and telophases. Also, Zn(2+) induced vacuolated nuclei and irregular prophases. The induction of chromosomal stickiness and chromosomal aberrations such as bridges and breaks indicates its action on the chromosome. These abnormalities (chromosome breaks and chromosomal bridges at ana-telophases) indicate true clastogenic potential of the ions tested.     

Protective roles of exogenous polyamines on chromosomal aberrations in <Emphasis Type="Italic">Hordeum vulgare</Emphasis> exposed to salinity

The effects of exogenous polyamines (PAs): spermine (Spm), spermidine (Spd), cadaverine (Cad) and putrescine (Put) on mitotic activity and chromosomal aberrations in root meristem cells of Hordeum vulgare L. (barley) seeds exposed to salinity were analyzed. The PAs significantly inhibited cell division in distilled water. Furthermore, most of these PAs (except for Spd) caused a significant increase in the frequency of chromosomal aberrations as compared to control group. Seeds treated with Put caused the highest percentage of mitotic abnormalities in total. The negative effect of salinity on mitotic index and the frequency of chromosomal aberrations increased with increasing salt concentration. PAs studied could not be successful in ameliorating of the negative effect of salinity on mitotic activity. Particularly, exposure to Cad and 0.40 M NaCl caused a complete block of cell division in total. However, most of the PA studied showed a perfectly performance in alleviating the detrimental effects of increasing salinity on chromosomal aberrations.     

Mitotic cycle kinetics of root meristems of isolated barley embryos and intact seedlings. Labelling of nuclei by3H- thymidine and its cytogenetic consequences

The duration of the mitotic cycle and its individual phases was estimated in root meristems of isolated barley embryos and intact barley seedlings by means of pulse labelling with³H-thymidine and construction of labelled mitoses curve. The duration of the whole mitotic cycle in the cell population of root meristems of isolated barley embryos cultivated in the aerated liquid complete medium is 12.2 h. The mitotic cycle time of root meristems of intact barley seedlings, oultived in Petri dishes on wet blotting paper is 9.2 h. Most of root meristem cells belong to the fraction of rapidly proliferating cells, but this fraction exerts a high degree of variability by itself. Pulse treatment by³H-thymidine in our experimental conditions (74 kBq ml^-1 - or 2 μCi ml^-1, exposure 0.5 h) did not induoe any chromosomal aberrations in unlabelled cells and only a very low frequency of chromosomal aberrations in labelled cells. Measuring the cell population kinetics by pulse labelling with³H-thymidine can be used simultaneously with the study of induction of ohromosomal aberrations by mutagens.     

Evaluation of malathion-induced cytogenetical effects and oxidative stress in plants using Allium test

The present study emphasized to explore the toxicity effect of malathion on plants using Allium test. The experiments explored the mitotic inhibition, growth and activity of antioxidant enzymes in roots of Allium cepa at different concentrations (50, 125, 250 and 375 ppm) of malathion under different exposure periods (3, 9 and 18 h). The results revealed that all concentrations of malathion were capable to decline the root growth. Malathion-induced mitotic alterations varying from reduction in mitotic index (MI), relative division rate (RDR) and phase distribution along with large number of chromosomal aberrations. These changes were of varying degree depending on the concentration and treatment period. The roots treated with malathion (375 ppm) showed significant (p ≤ 0.05) higher levels of superoxide dismutase and catalase than the control, while the activity of peroxidase was significantly (p ≤ 0.05) low. At 375 ppm malathion, malondialdehyde content was significantly high (p ≤ 0.05) that was increased with the treatment period. Findings concluded that variations in mitotic index, chromosomal aberrations, alterations in malondialdehyde content and activities of antioxidant enzyme could serve as the useful indicators for monitoring the effects of malathion exposures in the real scenario. Superoxide dismutase and catalase enzyme play vital roles in the antioxidant defence mechanisms under malathion toxicity. According to the data on the malondialdehyde content show malathion to be capable of producing superoxide radicals indirectly, and to result in membrane damage and oxidative stress.     

The effect of lead on Allium cepa L.

The effect of lead on Allium cepa L. at concentrations of 0.1, 1.0, 10, 50, 100 and 200 ppm were studied. Analysis focused on root growth, frequency of mitosis in a meristematic zone, and chromosomal aberrations. It was observed that lead reduces root growth and the frequency of mitotic cells in meristematic zones, and increases the frequency of aberrant cells. The intensity of the effects is a function of lead concentration.     

Chromosome damage in rat pulmonary alveolar macrophages following ozone inhalation

To determine whether ozone is clastogenic at environmentally relevant exposure levels, rats were exposed for 6 h to 0.0, 0.12, 0.27, or 0.80 ppm ozone. The alveolar macrophages were isolated from animals sacrificed 28 h after the end of the exposure. The mitotic index and frequency of chromosome aberrations were determined. No change in the mitotic index was detected following 0.12 ppm ozone exposure. A significant decrease in mitotic index was observed after exposure to 0.27 ppm ozone; a significant (4-fold) increase in the frequency of dividing macrophages was detected following exposure to 0.8 ppm ozone. Only chromatid-type aberrations were observed. There was a significant increase in the frequency of cells with chromatid gaps and in the frequency of cells with chromatid deletions. Animals exposed to 0.27 ppm ozone had the highest proportion of cells with chromatid deletions (0.172) relative to background level (0.028). No exchanges or chromosome-type aberrations were detected in any of the animals. These data suggest that ozone, at relatively low levels, is clastogenic in macrophages from exposed rats.     

Chromotoxic effects of exogenous hydrogen peroxide (H<Subscript>2</Subscript>O<Subscript>2</Subscript>) in barley seeds exposed to salt stress

The effect of exogenous hydrogen peroxide (H₂O₂) on mitotic activity and chromosomal aberrations in root tip meristems of barley (Hordeum vulgare L. var. Tokak 157/37) germinated under salinity was analyzed. The inhibitory effect of salinity on mitotic index and the frequency of chromosomal aberrations increased with increasing salt concentration (0.00 control, 0.35, 0.40, 0.45 M, molal NaCl). The frequency of chromosomal aberrations of seeds germinated in medium with 0.40 M NaCl after pretreatment with H₂O₂ (30 μM, micromolal) was significantly higher than the control group. The highest concentration of NaCl (0.45 M) together with H₂O₂ caused total inhibition of germination. In this study, the intention was to determine the performance of H₂O₂ in alleviating detrimental effect of salt stress on mitotic activity and chromosomal aberrations. However, H₂O₂ did not reduce the detrimental effect of NaCl on these parameters. Also, it caused higher chromotoxic effect compared to those of control groups.     

The influence of exogenous DNA of different origin on the mitosis and chromosomes of irradiated meristematic cells ofVicia faba

In this paper we compare the influence of heterologous and isologous DNA on the radiation damage repair of primary root meristematic cells ofVicia faba. Roots, irradiated by exposure of 150 r were cultivated at different time intervals either in tap water, or in a solution of heterologous or isologous DNA. In comparing mitotic activity of meristematic cells it was found that both types of DNA studied enhance the recovery of irradiated cells. The frequency of postmetaphase chromosomal aberrations of irradiated cells was influenced also by post-irradiation action of exogenous DNA. While heterologous DNA exhibited synergical effect with radiation in the sense that it increased the post-irradiation incidence of aberrations in all time intervals studied, isologous DNA had a strong repair effect—the application caused a significant decrease of the percentage of post-metaphase aberrations. Both kinds of DNA caused changes in the relation of chromosome to chromatid aberrations; a higher percentage of chromatid aberrations was registered. The study of the distribution of aberrations between large and small chromosomes ofVicia faba showed that the post-irradiation application of heterologous DNA increases damage of small chromosomes while isologous DNA caused an increased repair ability in this chromosomal group.     

Role of some growth regulators on cytogenetic activity of barley under salt stress

The effects of gibberellic acid (GA₃), kinetin (KIN), benzyladenine and ethylene (E) on mitotic activity and chromosomal aberrations in root tips of barley seeds (Hordeum vulgare L. cv. “Bülbül 89”) germinated under salt stress were investigated. It was determined that all of these plant growth regulators (PGRs) decreased mitotic index in root tips of barley seeds germinated at 20 °C and in distilled water. Furthermore, some of the PGRs studied increased significantly the frequency of chromosomal aberrations. The frequency of chromosomal aberrations in seeds treated with E and KIN was considerably higher than in the seeds germinated under nonstress conditions. The inhibitory effect of salt stress on mitotic index increased with increasing salt concentration (0.30, 0.35, 0.40 and 0.45 molal, m). GA₃ and KIN pretreatments showed a successful performance in ameliorating the negative effects of increasing salinity on mitotic activity. The number of chromosomal aberrations also increased with increasing NaCl concentration. However, most of the PGR pretreatments studied alleviated the detrimental effects of increasing salinity on chromosomal aberrations. KIN pretreatment at 0.30 and 0.35 m salinity could not rescued the cytogenetic activity of salt stress on this parameter.     

Evaluating cadmium toxicity in the root meristem of <Emphasis Type="Italic">Pisum</Emphasis><Emphasis Type="Italic">sativum</Emphasis> L.

The effect of cadmium (Cd) was studied on root tips of Pisum sativum L. Seeds of P. sativum were treated with a series of concentrations ranging from 0.125, 0.250, 0.500 and 1.000 mM for 6 h. The effect of Cd was analyzed by studying the percentage seed germination, radicle length (RL), mitotic index (MI) and chromosomal aberrations (CAs) in root tip. The results revealed that Cd had significant impeding effect on the root meristem activity of P. sativum at 0.500 and 1.000 mM as noticed by reduction in seed germination percentage and RL compared to control. Furthermore, it also reduced MI in dose-related manner compared to control. Additionally, the variation in the percentage of mitotic abnormalities was observed. The overall percentage of aberrations generally increased with increasing concentrations of Cd. Among these abnormalities laggards, bridges, stickiness, precocious separation and fragments were most common. The obtained results demonstrated that the Cd treatment leads to a significant reduction in MI and increase in CAs. Overall results allow us to suggest that the Cd has clastogenic effect on the crop.     

Chromosome Aberrations and Ageing Root Meristems

Changes in mitotic activity and frequency of spontaneous chromosomeaberrations have been determined for primary roots of Zea muys,Pisum sativum, Vicia faba var. green Windsor and var. minor,and Allium cepa. Mitotic activity was found to decline withage in all root apices. In the primary root apices immediately after germination, therewas a high frequency of spontaneous chromosome aberrations whichdeclined with time after germination. However, In the case ofV. faba var. green Windsor, Z mays and P. sativum there wasa return to the high levels of spontaneous chromosome aberrationon ageing of the root apex. root meristems, mitotic activity, chromosome aberrations, ageing     

Cytotoxic and genotoxic effects of dioxacarb by human peripheral blood lymphocytes CAs and Allium test

Dioxacarb (Elecron, Famid) is a phenyl methylcarbamate insecticide and in vitro cytotoxic and genotoxic effects of this pesticide on human peripheral blood lymphocytes and Allium root meristematic cells were investigated by chromosomal aberrations (CAs) and Allium test. Human lymphocytes were treated with 62.5, 125, 250 and 500 ppm doses of dioxacarb for CAs. CA/cell, abnormal cell % and mitotic index % (MI %) data were obtained from these concentrations in 24 and 48 h treatment periods. Dioxacarb did not increase the CA/cell frequency significantly, so this insecticide was not identified as genotoxic. But it was found cytotoxic especially at 250 and 500 ppm concentrations because of the reduced the MI % and increased the abnormal cell %. In Allium test, 25 ppm (EC50/2), 50 ppm (EC50) and 100 ppm (EC50 × 2) concentrations were used for root growth inhibition (EC50 determination) and Allium mitotic index (MI) determination tests. The used concentrations of dioxacarb induced dose-dependent inhibition of MI and root growth on root meristems. Mitotic inhibition of dioxacarb was found significantly higher than for the positive control. These Allium results indicated the high cytotoxicity of dioxacarb. The present study is the first research on cytotoxicity and genotoxicity of dioxacarb by human lymphocyte CAs and Allium test.     

The influence of organic and inorganic chelators on the toxicity of bulk and nanoparticles of zinc oxide during germination and seedling growth of Nicotiana tabacum L.

Nano and bulk-forms of zinc oxide (ZnO) are used extensively in industry and consequently may accumulate in the environment. However, there is little information available on the comparative effects of these forms during the critical early stages of plant life. Furthermore, the role of chelating agents, which affect the bioavailability of metals, in ameliorating plant stress due to exposure to nano and bulk-forms of ZnO is not well characterised. In this study, the effects of different concentrations (0.5, 2.5, 5, 10, 50 and 100 ppm) of nano ZnO (22 nm) and bulk ZnO (natural form, 1000 nm) with and without organic (citrate) and inorganic (EDTA) chelators on germination and seedling growth, and oxidative stress markers of Nicotiana tabacum L. were compared. Chelators (without ZnO) enhanced root growth, whilst ZnO negatively affected seedling growth. ZnO toxicity was often mitigated by adding chelators, especially citrate, although at the highest levels (50 and 100?ppm) of ZnO, toxicity was more pronounced when chelated with EDTA, but was decreased with citrate. Collectively, our findings provide important information regarding the different morpho-physiological and biochemical effects of bulk and nano ZnO and organic and inorganic chelators (citrate and EDTA), which are all prevalent in the environment.     

Significato dei Tegumenti in Cariossidi di Triticum Durum Desf. Trattate Con Raggi X

Abstract

Significance of integuments in caryopses of « Triticum durum » Desf. treated with X-rays. Experiments were carried out on Triticum durum « Cappelli », to test the influence of seed coats on the damage by X-rays. The damage was evaluated in terms of germination, seedling growth and chromosomal aberrations. Seeds in which the embryo was covered (with pericarp and spermoderm), uncovered (without pericarp) and bared (without pericarp and spermoderm), were irradiated at the total exposure of 1000, 2000 and 4000 R of X-rays. The growth of seedlings coming from covered and uncovered embryos, X-rayed at 1000 R, results better than that of bared embryos. The chromosomal aberrations analysis carried out in root meristem cells at anaphase, during the first mitotic cycle, show the highest damage in seedlings coming from uncovered and bared embryos and the type of aberrations was almost chromatidic (B'). We therefore forward the hypothesis that the removal of the integuments (pericarp and spermoderm) could possibly accelerate the process of germination, and could also produce in the embryo a greater sensitivity to X-rays.     

HRS1 acts as a negative regulator of abscisic acid signaling to promote timely germination of Arabidopsis seeds

In this work, we conducted functional analysis of Arabidopsis HRS1 gene in order to provide new insights into the mechanisms governing seed germination. Compared with wild type (WT) control, HRS1 knockout mutant (hrs1-1) exhibited significant germination delays on either normal medium or those supplemented with abscisic acid (ABA) or sodium chloride (NaCl), with the magnitude of the delay being substantially larger on the latter media. The hypersensitivity of hrs1-1 germination to ABA and NaCl required ABI3, ABI4 and ABI5, and was aggravated in the double mutant hrs1-1abi1-2 and triple mutant hrs1-1hab1-1abi1-2, indicating that HRS1 acts as a negative regulator of ABA signaling during seed germination. Consistent with this notion, HRS1 expression was found in the embryo axis, and was regulated both temporally and spatially, during seed germination. Further analysis showed that the delay of hrs1-1 germination under normal conditions was associated with reduction in the elongation of the cells located in the lower hypocotyl (LH) and transition zone (TZ) of embryo axis. Interestingly, the germination rate of hrs1-1 was more severely reduced by the inhibitor of cell elongation, and more significantly decreased by the suppressors of plasmalemma H(+)-ATPase activity, than that of WT control. The plasmalemma H(+)-ATPase activity in the germinating seeds of hrs1-1 was substantially lower than that exhibited by WT control, and fusicoccin, an activator of this pump, corrected the transient germination delay of hrs1-1. Together, our data suggest that HRS1 may be needed for suppressing ABA signaling in germinating embryo axis, which promotes the timely germination of Arabidopsis seeds probably by facilitating the proper function of plasmalemma H(+)-ATPase and the efficient elongation of LH and TZ cells.     

Cytogenetic and microtubule array effects of the zineb-containing commercial fungicide formulation Azzurro(®) on meristematic root cells of Allium cepa L

Zineb [ethylene bis(dithiocarbamate) zinc] is a widely employed foliar fungicide for agricultural and industrial applications. Allium cepa L. is a reliable model for the assessment of xenobiotic genotoxicity and cytotoxicity. We evaluated the effects of the zineb-containing commercial formulation Azzurro(?) (70% zineb) in cell cycle stages of the meristem root cells of A. cepa. The mitotic index (MI), chromosomal aberrations at anaphase/telophase (CAs), micronuclei (MN), and abnormalities in immunodetected microtubule structures, e.g., preprophasic band (PPB), mitotic spindle (MS), and phragmoplast (Phrag), were used as end-points. Azzurro(?) (1 and 10μg/ml) induced a significant increase in the frequency of CAs (P<0.05), and the higher concentration inhibited the MI (P<0.05) compared to control values. The frequency of MN did not differ from control values at any concentration. Treatment with 1μg/ml Azzurro(?) induced a significant increase in the frequency of abnormal PPB (P<0.01), MS (P<0.001), and Phrag (P<0.01) and, at 10μg/ml, enhancements in the frequencies of abnormal MS (P<0.05) and Phrag (P<0.05) were seen. A tubulin immunodetection assay showed that exposure to Azzurro(?) interferes with normal assembly of microtubule structures during mitosis.     

Cytogenetic effects of pulsing electromagnetic field on human lymphocytes in vitro: chromosome aberrations, sister-chromatid exchanges and cell kinetics

Exposure of human lymphocyte cultures to a pulsing electromagnetic field (PEMF; 50 Hz, 1.05 mT) for various durations (24, 48 and 72 h) resulted in a statistically significant suppression of mitotic activity and a higher incidence of chromosomal aberrations. Furthermore, the shorter exposure times (24 and 48 h) did not cause a significant delay in cell turnover (cell proliferation index) or an increase in the baseline frequency of sister-chromatid exchanges (SCE). However, cultures continuously exposed to PEMF for 72 h exhibited significant reduction of the cell proliferation index (CPI) and an elevation of SCE rate. These results suggest that exposure to PEMF may induce a type of DNA lesions that lead to chromosomal aberrations and cell death but not to SCE, except probably at longer exposure times.     

Assessment of Nanotoxicity (Cadmium Sulphide and Copper Oxide) Using Cytogenetical Parameters in <Emphasis Type="Italic">Coriandrum sativum</Emphasis> L. (Apiaceae)

Present investigation deals with cytogenetical consequences (using attributes namely, seed germination, seedling length, mitotic index, mitotic and meiotic abnormalities and pollen grain sterilities) of cadmium sulphide (CdS) and copper oxide (CuO) nanoparticles (NPs) treatment in Coriandrum sativum L. (Family: Apiaceae, spice of commerce). Ethyl methanesulphonate (EMS), a conventional mutagen is used as positive control. Results suggest that both CdS- and CuO-NPs can induce growth inhibition and cause cytological aberrations in both mitotic and meiotic cells in the studied species. EMS also responds similarly as that of NPs. Study highlights that rooted plant species can be effectively used as model for assessment of nanotoxicity considering cytogenetical parameters.     

Bioremediation of municipal sludge by vermitechnology and toxicity assessment by Allium cepa

The aim of this study was to evaluate municipal sludge (MS) for its toxic potential by Allium cepa and also to understand the effect of vermicomposting on the reduction of toxicity, if any. Municipal sludge (MS) and vermicomposted sludge (VS) were evaluated. Elemental analysis of MS showed the presence of heavy metals. Morphological studies of A. cepa roots indicated coiled and wavy roots on exposure to MS but no root abnormality was reported in VS. Under genotoxic studies, inhibition in mitotic index was concentration dependent and the control values of 11.76 gradually reduced to 5.40 at 10% MS leachate whereas mitotic index was increased to 9.48 at 10% VS leachate. Exposure of leachate induced chromosomal aberrations, micronucleus formation and binucleate cells in a dose dependent manner. However, mitotic aberrations were observed significant at 10% MS leachate but they were insignificant at 10% VS leachate. The wet and dry weight of roots, root elongation and chlorophyll contents were reduced as the concentration of leachate increased but VS leachate did not produce considerable reduction. The wet and dry weight of A. cepa roots were 20.312 g and 3.250 g respectively and they were reduced to 10.82 g and 1.68 g respectively at 10% MS leachate but VS leachate showed an increase to 18.127 g and 2.53 g respectively. Total chlorophyll in control, 10% MS leachate and 10% VS leachate were 0.245 g, 0.162 g and 0.214 g respectively. It could be concluded that the MS was toxic to a remarkable extent but vermicomposting of sludge might be beneficial for bioremediation and recommended before land filling.     

Genotoxic effects of the pesticides Rubigan, Omite and Rovral in root-meristem cells of Crepis capillaris L

Three pesticides have been studied for their genotoxicity by the use of assays in the plant Crepis capillaris, aimed at measuring chromosomal aberrations, micronuclei and sister chromosome exchange (SCE). The fungicides Rubigan 12 EC (fenarimol) and Rovral 25 Flo (iprodione) and the insecticide Omite 57 E (propargite) are all widely used nowadays. The aim of our study was to evaluate the genotoxic effects of these pesticides at concentrations corresponding to those applied in agricultural practice. In preliminary experiments we found that these concentrations do not influence cell proliferation and do not inhibit the growth of root meristems. In all experiments formulated commercial products were used. From the results we conclude that the three pesticides did not induce chromosomal aberrations as estimated by metaphase and anaphase analyses. They were also not capable to induce SCE. Rubigan did not induce micronucleus formation even at the highest concentration tested, but Omite and Rovral markedly increased micronucleus formation. The MN response depended on the sampling time and the concentration used, which showed a significant dose-response correlation (r=0.978, P<0.01 and r=0.941, P<0.01, respectively). A greater increase in micronucleus frequency was observed after Rovral treatment, where the highest concentration gave a response 8-10-fold above the negative control. Both pesticides induced high frequencies of lagging chromosomes, even after exposure to the lower test concentrations. The presence of lagging chromosomes is an indication of anti-microtubule activity of the pesticides tested. This effect was more strongly expressed after exposure to the two higher concentrations of Omite and Rovral. In this case a complete destruction of the mitotic spindle was observed, resulting in C-mitoses as well as in numerical aberrations-polyploidy and aneuploidy. The present findings suggest that Omite and Rovral at concentrations comparable to those used in practice can be regarded as potential aneugens.