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Evaluation of cytogenetic effects of isoproturon on the root meristem cells of Allium sativum.

OBJECTIVE: To validate the use of Allium sativum as a reliable test model for genotoxicity, isoproturon, a substituted phenylalkylurea herbicide, was evaluated on the root meristem cells by this plant system. METHOD: Test concentrations were selected by determining EC50 and root tips were exposed to various concentrations for 6 or 24 hr. EC50 concentration was calculated to be 70. 8 ppm for the root growth. In addition to root growth retardation exposure to isoproturon induced morphological changes like discolouration and stiffness of roots. RESULTS: Exposure to various experimental concentrations of isoproturon (35-280 ppm), including EC50, significantly and dose-dependently inhibited the mitotic index and induced chromosome breaks/mitotic aberrations at 6 or 24 hr. CONCLUSION: The frequency of aberrations was found to be decreased in a dose dependent manner at 24 or 48 hr post exposure, however in comparison of control cells the frequency of aberrations was considerably high which indicates genotoxicity potentials of isoproturon. Further, present study also suggests that Allium sativum is a sensitive, efficient, and reliable test system for measuring the genotoxicity potential of environmental chemicals.

Chromosome Aberrations↗

[Genotoxicity evaluation of paracetamol applying Allium test].

Paracetamol genotoxic potential was evaluated among different concentrations, applying Allium test on Allium cepa. Total number of roots, number of dark roots, the length of the longest root, the average root, and the shortest root were determined. Statistically significant differences among total number of roots (p > 0.05) was observed at concentrations of 50 microg/ml and 100 microg/ml, and highly statistically significant differences at concentrations of 5 microg/ml and 25 microg/ml, while at the highest concentration (400 microg/ml) was observed statistically significant higher number of roots in comparation to all other concentrations of paracetamol and control group. The results of research suggest the concentrations of 5 microg/ml, 25 microg/ml and 400 microg/ml for further evaluation of paracetamol mutagenic potential.

Acetaminophen↗

[Phytochemical and microbiological characterization of two Allium cepa L. extracts in order to include in dermo-cosmetics].

We have obtained two Allium cepa extracts (C1 and C2) which were characterized from chemical and microbiological point of view. The C1 extract is the richest concerning the content in flavonoids, triterpenic acids, amino acids, compounds recognized for their beneficial effects in wound healing. All this data shows us the possibility of using the Allium cepa extracts in the treatment of wounds, scars. As well, the antimicrobial activity of the two extracts was evaluated. The C2 extract was efficient as antimicrobial agent, but there are necessary special preserving conditions.

Amino Acids↗

RBE of d(50)-Be neutrons for induction of chromosome aberrations in Allium cepa onion roots.

RBE/absorbed dose relationship of d(50)-Be neutrons was determined for the induction of chromosome aberrations in Allium cepa onion roots. Neutrons are produced at the cyclotron "Cyclone" by bombarding a thick beryllium target with 50 MeV deuterons. Two biological criteria were selected: (1) mean number of aberrations (mainly breaks) per cell in anaphase and telophase, (2) fraction of intact cells in anaphase and telophase. For the two criteria, RBE increases continuously from about 7 to 12 as the neutron absorbed dose decreases from 0.4 to 0.1 Gy. RBE values for the first criterion are slightly higher than for the second one. This observation is interpreted in terms of the analysis of the distribution of the aberrations in the cells. In logarithmic coordinates, RBE/absorbed dose relationships for the two criteria are almost linear with a slope close to -1/2. RBE values observed for induction of chromosome aberrations in Allium cepa are higher than those generally observed for biological effects related to mammalian cell lethality.

Beryllium↗

Inhibitory effect of adrenaline and hydrocortisone on the growth of Allium cepa roots.

The roots of Allium cepa were allowed to grow in distilled water containing 10(-4) M adrenaline hydrochloride or 2 X 10(-3) M hydrocortisone sodium succinate. Adrenaline inhibited the growth of roots; they decreased in length, number and total dry weight. The total amount of DNA in the roots was reduced much less than that of extracellular root components after adrenaline. Also hydrocortisone treatment resulted in a considerable decrease of the length and dry weight of Allium cepa roots. Both DNA and extracellular root components were influenced.

Cell Division↗

The effect of Allium sp. on the extension of lipolysis and proteolysis in Van herby cheese during maturation.

The aim of this study was to determine the effect of herb (Allium sp.) on biochemical changes of herby cheese produced in Turkey. Raw cows' milk was used for cheese manufacture. Five groups of cheeses, containing 0 (as control), 0.5, 1, 2, and 3% herb, were produced and coded as K, A, B, C, and D respectively. All cheese groups were ripened at 8 degrees C for 90 days. Samples were taken from cheeses after 3, 15, 30, 60 and 90 days, and analysed for lipolysis (as acid degree value) and proteolysis (water-soluble nitrogen, TCA-soluble nitrogen, and PTA-soluble nitrogen). It was found that lipolysis in herby cheeses increased with increasing herb addition, and the increase in lipolysis degree was significant (P < 0.05) in cheese D. Water-soluble N, TCA-soluble N, and PTA-soluble N as indicator of proteolysis degrees were affected significantly (P < 0.05) by increasing herb ratios.

Allium↗

Antioxidative activity and ameliorative effects of memory impairment of sulfur-containing compounds in Allium species.

The antioxidative activity and ameliorative effects on memory impairment by sulfur-containing compounds which occur in Allium vegetables such as onion and garlic were investigated. The antioxidative activities of S-alk(en)yl-L-cysteines and their sulfoxides, volatile alk(en)yl disulfides and trisulfides, and vinyldithiins were examined by using human low-density lipoprotein. It was elucidated that the alk(en)yl substituents and the number of sulfur atoms in the compounds were important for the antioxidative activities. To demonstrate the ameliorative effects on memory impairment, onion extract and synthesized di-n-propyl trisulfide were administered to senescence-accelerated mouse P8. The behavioral experiments showed that onion extract and di-n-propyl trisulfide had highly ameliorative effect of memory impairment. Furthermore, it was found that the hippocampus lipid hydroperoxide in senescence-accelerated mouse P8 was decreased by the administration of di-n-propyl trisulfide. These results suggest that di-n-propyl trisulfide contained in onion ameliorates memory impairment in SAMP8 mouse by its antioxidant effect.

Aging↗

Scavenger activity of Allium psekemense B. Fedtsch.

In the present study the antioxidative properties of the bulbs, leaves and stalks of Allium psekemense B. Fedtsch were investigated. The activities of antioxidant enzymes (superoxide dismutase, catalase, peroxidase, glutathione peroxidase), quantities of malonyldialdehyde, superoxide and hydroxyl radicals and reduced glutathione and also the content of total flavonoids, chlorophylls a and b, carotenoids, vitamin C and soluble proteins were determined. The results indicate that extracts from all plant organs exhibited antioxidant activity. The highest antioxidant activity was observed in the leaves. Furthermore, the ESR signal of DMPO-OH radical adducts in the presence of the leaf phosphate buffer (pH 7) extract was reduced by 54.3%.

Allium↗

Purification and characterization of a mannose-specific lectin from Shallot (Allium ascalonicum) bulbs.

A new mannose-binding lectin was isolated from shallot (Allium ascalonicum) bulbs by affinity chromatography on an immobilized D-mannose column. The lectin (A. ascalonicum agglutinin, AAA) appeared homogeneous by polyacrylamide gel electrophoresis at pH 4.3 and gave a single protein band with an apparent M(r) of 11 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and a single symmetrical peak of 11 kDa by gel filtration on a Sephacryl S-200 HR column, indicating that AAA exists as a monomeric protein at neutral pH under the gel filtration condition employed. However, chemical cross-linking studies revealed that some degree of self-association of the lectin molecules occurs and that the lectin exists in solution as a mixture of monomers and oligomers. Scatchard analysis of equilibrium dialysis data showed the presence of one carbohydrate binding site for Man (alpha 1-3) Man-alpha-O-Me per monomer, with Ka = 1.62 x 10(4) M-1. The carbohydrate-binding properties of the purified AAA were investigated by quantitative precipitation and hapten inhibition assays. Purified AAA precipitated asialofetuin, asialotransferrin, asialothyroglobulin, asialoorosomucoid, as well as their agalacto derivatives, but did not precipitate either sialylated glycoproteins or mucins. AAA also reacted strongly with the highly branched yeast mannan obtained from Saccharomyces cerevisiae. Of the monosaccharides tested only D-mannose was a hapten inhibitor of the AAA-asialofetuin precipitation system, whereas D-glucose, D-altrose, D-talose, N-acetyl-D-mannosamine, and derivatives of D-mannose, including 2-deoxy-, 2-deoxy-2-fluoro-, 3-deoxy-, and 6-deoxy-D-mannose were noninhibitors. These results suggest that the presence of equatorial hydroxyl groups at the C-3 and C-4 positions, an axial hydroxyl group at the C-2 position, and a free hydroxyl group at the C-6 position of the pyranose ring are the most important loci for the binding of D-mannose to AAA. Of the oligosaccharides tested, the best inhibitors were oligosaccharides containing terminal Man(alpha 1-6) [Man(alpha 1-3)]Man groups. Oligosaccharides containing either Man(alpha 1-3)Man or Man(alpha 1-6)Man units were also moderately good inhibitors of the AAA-asialofetuin precipitation system. These results indicate that AAA has an extended carbohydrate-binding site, which is most complementary to a branched mannotriosyl residue, i.e., Man(alpha 1-6)[Man(alpha 1-3)]Man.(ABSTRACT TRUNCATED AT 400 WORDS)

Allium↗

Monoclonal antibody raised against human mitotic cyclin B1, identifies cyclin B-like mitotic proteins in synchronized onion (Allium cepa L.) root meristem.

Cyclin B-like mitotic proteins have been detected in synchronized Allium cepa L. root tip cells by using mouse monoclonal anti-cyclin B1 antibody raised against human cyclin B1. Immunoblot shows two closely placed isoforms of cyclin B-like proteins having an apparent molecular weight around 54 kDa. In vivo [35S]-methionine labelling followed by immunoprecipitation and autoradiography indicates that cyclin B-like proteins are mainly synthesized in the G2 phase of the cell cycle and destroyed in late mitosis. Immunoblotting data depict that the level of cyclin B-like proteins reaches the maximum at the late G2 to early M phase; and it becomes degraded in the late hours of mitosis. Moreover, the cyclin B isoforms are stabilized in colchicine-arrested metaphase cells as already reported in animal cells.

Allium↗

Reorganization and condensation of chromatin in mitotic prophase nuclei of Allium cepa.

This paper studies the process and features of chromosome construction in mitotic prophase cells of Allium cepa. The results showed that a prominent reorganization of chromatin occurred during G2--early prophase. The 250-400 nm thick compact chromatin threads in G2 nuclei began to disorganize into about 30, 100 and 220 nm chromatin fibres which constituted the loosely organized chromosome outlines in early prophase before chromosome condensation. In middle prophase, chromosome condensation was characterized by the formation of many condensed regions (aggregates of chromatin), which increased in size (1-1.5 microns) when prophase proceeded. Meanwhile, the chromatin threads that constituted and connected the condensed regions became increasingly thicker (120-250 nm). In late prophase adjacent condensed regions fused to form cylinder-shaped chromosomes. Based on these observations, we come to the conclusion that the construction of prophase chromosomes is a two-step process, that is, the reorganization and condensation of chromatin. In addition, we report the study of silver-stained, DNA- and histone-depleted prophase chromosomes, describe morphological features of the non-histone protein (NHP) residue in early, middle and late prophase chromosomes, and discuss the roles of NHPs in chromosome construction.

Allium↗

The spatial arrangement of Allium triquetrum chain quadrivalents at metaphase I as reviewed by confocal microscopy.

We examined the three-dimensional arrangement of bivalents and, in particular, a chain of four chromosomes (chain quadrivalent) in the metaphase I spindle of pollen mother cells of Allium triquetrum by confocal microscopy. Firstly, we show by optical sectioning and three-dimensional image reconstruction that the cooriented pairs of centromeres of all seven bivalents lie virtually parallel to each other in the metaphase I spindle, parallel to the long axis of the spindle. Secondly, we likewise show that the four centromeres of the chain quadrivalent are aligned in the metaphase I spindle in, essentially, a two-dimensional array, not in a three-dimensional array, as proposed by some other authors. This two-dimensionality has its basis, we argue, in the principle that poleward directed spindle forces minimise centromere-to-pole distances and therefore align pairs of centromeres connected to opposite poles most axially (vertically) in the spindle. These distances are minimised for the quadrivalent as a whole only when it lies in two dimensions, i.e. in a plane parallel to the spindle axis.

Allium↗

rDNA site number polymorphism and NOR inactivation in natural populations of Allium schoenoprasum.

Nucleolar-organiser activity has been studied by silver staining and by in situ hybridization with an rDNA probe in two populations of Allium schoenoprasum. One population is monomorphic with NORs and rDNA sites terminal on the short arm of pair 8 in all individuals. The other populations is monomorphic for pair 8 NORs but is also polymorphic for NORs on the long arm of pair 7. All plants in this population carry ribosomal cistrons on both chromosomes of pair 7 but 0, 1 or 2 of these sites can be active in rRNA synthesis. Cis-acting nucleolar-suppression affects the pair 7 locus. We suggest that there has been progressive reduction in the number of NORs during the evolution of A. schoenoprasum.

Allium↗

The Ty1-copia group retrotransposons of Allium cepa are distributed throughout the chromosomes but are enriched in the terminal heterochromatin.

The genomic organization and diversity of the Ty1-copia group retrotransposons has been investigated in a monocotyledonous plant, Allium cepa. We used the polymerase chain reaction (PCR) to generate sequences corresponding to a conserved domain of the reverse transcriptase gene of Ty1-copia retrotransposons in this plant. Sequence analysis of 27 of these PCR products shows that they are a highly heterogeneous population, a feature which is common in plants but not in yeast and Drosophila. Slot-blot analysis shows there are 100,000-200,000 copies of Ty1-copia group retrotransposons within the A. cepa genome (2C = 31.7 pg), indicating that they are a significant component of the genome of this plant. In situ hybridization to metaphase chromosomes reveals that Ty1-copia retrotransposons are distributed throughout the euchromatin of all chromosomes of A. cepa but are enriched in the terminal heterochromatic regions, which contain tandem arrays of satellite sequences. This is the first clear evidence for the presence of Ty1-copia retrotransposons in the terminal heterochromatin of plants and contrasts with the distribution of these elements in other plant species.

Allium↗

The inheritance of B chromosomes in Allium schoenoprasum L.

The inheritance of B chromosomes has been investigated. In Allium schoenoprasum from the River Wye, Powys; controlled crosses between plants of known B chromosome constitution were carried out, and the numbers of Bs present in the progenies scored. There was considerable heterogeneity in B chromosome transmission rates across the range of B chromosome numbers, through both the male and female lines, and in crosses involving odd and even numbers of Bs. The mean B number of the progeny was lower than the Mendelian expectation in 41 of the 57 crosses, significantly so in about a third of cases. No progeny had a mean B number significantly higher than expected. The mean transmission rate per B was not significantly different between parents with different numbers of Bs, in either the male or female line, or between plants carrying odd or even numbers of Bs. Transmission through the male line (0.43) was higher than through the female line (0.35), but not significantly so. The overall mean transmission rate per B (0.4), estimated from all crosses, was significantly lower than the expected rate of 0.5. This study provides conclusive evidence that B chromosomes in A. schoenoprasum from the River Wye lack an effective accumulation mechanism and, furthermore, show significant losses during transmission to the progeny of controlled crosses.

Allium↗

Subcellular localization of Cd in the root cells of Allium sativum by electron energy loss spectroscopy.

The ultrastructural investigation of the root cells of Allium sativum L. exposed to three different concentrations of Cd (100 mM, 1 mM and 10 mM) for 9 days was carried out. The results showed that Cd induced several significant ultrastructural changes high vacuolization in cytoplasm, deposition of electron-dense material in vacuoles and nucleoli and increment of disintegrated organelles. Data from electron energy loss spectroscopy (EELS) revealed that Cd was localized in the electron-dense precipitates in the root cells treated with 10 mM Cd. High amounts of Cd were mainly accumulated in the vacuoles and nucleoli of cortical cells in differentiating and mature root tissues. The mechanisms of detoxification and tolerance of Cd are briefly explained.

Allium↗

Constituents and the antitumor principle of Allium victorialis var. platyphyllum.

To search for cytotoxic components from Allium victorialis, MTT assays on each extract and an isolated component, gitogenin 3-O-lycotetroside, were performed against cancer cell lines. Cytotoxicities of most extract were shown to be comparatively weak, though IC50 values of CHCl3 fraction was found to be <31.3-368.4 microg/ml. From the incubated methanol extract at 36 degrees C, eleven kinds of organosulfuric flavours were predictable by GC-MS performance. The most abundant peak was revealed to be 2-vinyl-4H-1,3-dithiin (1) by its mass spectrum. Further, this extract showed significant cytotoxicities toward cancer cell lies. Silica gel column chromatography of the n-butanol fraction led to the isolation of gitogenin 3-O-lycotetroside (3) along with astragalin (4) and kaempferol 3, 4'-di-O-beta-D-glucoside (5). This steroidal saponin exhibited significant cytotoxic activities (IC50, 6.51-36.5 microg/ml) over several cancer cell lines. When compound 3 was incubated for 24 h with human intestinal bacteria, a major metabolite was produced and then isolated by silica gel column chromatography. By examining parent- and prominent ion peak in FAB-MS spectrum of the metabolite, the structure was speculated not to be any of prosapogenins of 3, suggesting that spiroketal ring were labile to the bacterial reaction. These suggest that disulfides produced secondarily are the antitumor principles.

Allium↗

Chromosomal location of a pollen fertility-restoring gene, Rf, for CMS in Japanese bunching onion (Allium fistulosum L.) possessing the cytoplasm of A. galanthum Kar. et Kir. revealed by genomic in situ hybridization.

In a previous study, we developed cytoplasmic male sterile lines of Allium fistulosum possessing the cytoplasm of A. galanthum, a wild species, by continuous backcrossing. Furthermore, we reported the presence of a pollen fertility-restoring gene (Rf) for cytoplasmic male sterility (CMS) in A. fistulosum from segregation of pollen fertility of backcross progenies. In the present study, genomic in situ hybridization (GISH), using genomic DNA of A. galanthum as the probe DNA and that of A. fistulosum as the blocking DNA, was applied to F(1) hybrids between both species and backcross progenies to determine the chromosomal location of the Rf locus. By means of GISH, eight chromosomes from A. galanthum were clearly discriminated from those of A. fistulosum in the F(1) hybrids, and chromosome substitution process through continuous backcrossing was visualized. Interestingly, the chromosome region from A. galanthum, specific to male fertile plants, was detected in one chromosome of BC(4) to BC(7) generations. Based on the karyotype analysis of the male fertile plants, the chromosome was identified as the 5F chromosome. Our results confirm that the Rf locus is located on the 5F chromosome of the male fertile plants. This is the first report that identified the chromosomal location of the pollen fertility-restoring gene in A. fistulosum.

Allium↗