Mitodepressive effects ofChelidonium alkaloids on root tip cells ofAllium cepa L

A comparison of the effects of a water extract of the fresh vegetation apex ofChelidonium majus L. var.majus, with those of berberine sulphate and of chelidonine hydrochloride is reported. The water extract of fresh stems and leaves ofChelidonium majus and the berberine sulphate solution had marked mitodepressive effects on onion root tip cells. Chelidonine hydrochloride had similar but less marked effects. On the basis of the results obtained it can be assumed that the most active group of substances with cytostatic effects, hindering the cells from entering mitosis, are protoberberine bases contained in the latex ofChelidonium majus. Within the range of the investigated concentrations the extract of fresh stems and leaves was less toxic for the cells than berberine sulphate. The data ascertained provide evidence that the mechanism of the cytostatic action of chelidonine, of berberine, as well as of the extract of fresh stems and leaves ofChelidonium, majus differs from the mechanism of the action of colchicine. Of the testedChelidonium alkaloids only chelidonine produced a partial inactivation of the mitotic spindle.     

Cytological effects of papaverine hydrochloride on root tips ofAllium cepa L.

Papaverine hydrochloride, which is an alkaloid found in opium, inducedc-metaphase,c-anaphase, non-congression, diagonal anaphase, chromosome and chromatid bridges, lagging of chromosomes, micronuclei and multinucleated cells in root tips ofAllium cepa. It is the only one amongst the alkaloids of opium which caused a high increase in MI after a long period of treatment with some low concentrations.     

Effect of morphine sulphate on mitosis ofAllium cepa L. root tips

The cytological effect of one of the alkaloid of Opium, namely morphine sulphate onAllium cepa root tips from a qualitative and quantitative point of view were studied. It was found that morphine sulphate caused partial effect on spindle formation and also showed a mito-depressive effects particularly after long treatment.     

Differentiation of metaxylem cell line in the root ofAllium cepa L.

Summary The differentiation processes of the metaxylem cell line in the root ofAllium cepa are characterized by amplification phenomena of repetitive DNA sequences mainly localized in heterochromatic regions of metaphase chromosomes. Moreover, these sequences are heavily methylated. This paper presents additional results on variation in endogenous DNA methylation in different developing root segments. The results show that methylation is higher in apical meristematic cells than the differentiating segments; contrastingly, total RNA synthesis seems to be correlated with undermethylation. Addition of labelled methyl groups to DNA by eukaryotic methylase, DNA digestions with different restriction enzymes specific for methylated sites and HPLC analysis confirmed the above results. Moreover, variation in methylation levels during differentiation occur not only at the internal cytosine of the-CCG-sites, but also at external cytosine. Furthermore, methylation affects other sites containing the trinucleotides-CXG-. In conclusion, root differentiation inAllium cepa seems to be correlated with gene activation modulated by the methylation/demethylation of particular DNA sequences.     

On the properties of fluorescing compounds in guard and epidermal cells of Allium cepa L.

Onion guard cells, in contrast to those of Vicia and Pisum, do not require an alkaline treatment in order to fluoresce. Fluorescing compounds of Allium cepa L. were characterized using in-vivo microspectrophotometry; furthermore, invitro chemical analysis for epidermal tissue, intact guard and epidermal cells, and isolated guard-cell protoplasts was performed. The emission intensity (_max 520 nm) decreased when intact onion guard cells were excited with 436 nm light, but increased (_max 470 nm) when excited at 365 nm. This photodecomposition at 436 nm is typical of flavins or flavoproteins whereas an increase in fluorescence intensity with excitation at 365 nm may be explained by the presence of other substances. The presence of flavins could not be unambiguously confirmed from these results. Indeed, the absorption spectra of the vacuolar area of guard cells did not show the peak at 445 nm which is characteristic for flavins. Furthermore, there was no decrease of absorption at the excitation wavelengths of 440 and 330 nm. Since spectral data indicate the presence at high amounts of flavonoids in guard and epidermal cells, this may reduce the sensitivity for the detection of flavins in guard cells. Using thin-layer chromatography and high-performance liquid chromatography together with hydrolytic procedures, flavonol glycosides with kaempferol and quercetin as aglycones substituted with sulphate and glucuronate were identified. Further studies on guard-cell metabolism should consider the presence of flavonoids in stomata of onion and other plants.Abbreviations GCP guard-cell protoplast - HPLC high-performance liquid chromatography - TLC thin-layer chromatography     

Microtubules and coated vesicles in guard-cell protoplasts ofAllium cepa L.

Protoplasts were prepared from the guard cells ofA. cepa. Epidermal peels taken from expanding green leaves and largely free of mesophyll were treated with Cellulysin, and protoplasts were harvested after 18 h of digestion. That the protoplasts were derived from guard cells was ascertained from their characteristic vacuolar autofluorescence and from observations showing that all other epidermal cells are killed in the peeling procedure. The protoplasts proved to be a good system with which to view the cell cortex and inner surface of the plasmalemma. The lysis of cells adhering to polylysine-treated, Formvar-coated grids, followed by negative staining in uranyl acetate, showed that many microtubules normally present in ordered arrays in situ remain closely applied to the inner surface of the plasmalemma in protoplasts. In addition, numerous vesiculate elements including coated vesicles and/or pits are present amongst the microtubules. Similar vesicles are evident in thin sections of fixed, embedded guard cells and protoplasts. The significance of these structures in the cell cortex is discussed.     

The screening of the enzyme and isoenzyme patterns in seeds ofAllium cepa L.

The screening of enzyme patterns in seeds ofAllium cepa cv. Všetatská revealed the presence of the following enzymes: alcohol dehydrogenase, lactate dehyd ogenase, NAD+- and NADP+-glyceraldehyde-3-phosphate dehydrogenase, glucose-6-phosphate dehydrogenase NAD+- and NADP+-malate dehydrogenase, NADH₂- and NADPH₂-tetrazolium reductase catalase, Superoxide dismutase, acid and alkaline phosphatase, L-leucine aminopeptidase, glutamate dehydrogenase, non-specific esterase, and cholinesterase. Altogether 17 enzymes were detected in onion seeds, nine of which had more than three isoenzymes, NAD+-malate dehydrogenase had 8, and non-specific esterase 9 isoenzymes. The demonstration of cholinesterase and Superoxide dismutase activities is remarkable.     

The effect of lead on the cell cycle in the root meristem ofAllium cepa L.

Summary Allium cepa (L.) adventitious roots were treated with lead (2.5 mg of Pb²⁺ [from Pb(NO₃)₂] per dm³) for 30–72 h. The cell cycle was studied by pulse labeling with [³H]thymidine. Mitotic activity kinetics, occurrence of disturbed mitoses (c-mitoses), and level of DNA synthesis were examined. It was found that lead prolonged the cell cycle and that cells in two phases of the cycle, G₂ and S, differed in their sensitivity to lead. Cells in G₂ were more sensitive; lead lengthened their cycle by 216% and disturbed the course of cell division by causing c-mitoses. Cells in S phase were less sensitive. Their cell cycle was longer by 55%. They went through their G₂ phase without major disturbances, mitosis in these cells was normal. During treatment ofA. cepa with lead, its destructive effects on cells were exerted only during the first few hours (around 6 h) of incubation. That is when the inhibition of mitotic activity, numerous disturbances of cell division, a decline in the number of cells synthesizing DNA, and a lower level of DNA synthesis were observed. As the incubation continued, the above processes were found to return to normal. In the discussion, data are presented supporting the hypothesis that during the initial period of exposure ofA. cepa to lead, this metal enters both the root apoplast and symplast, exerting a destructive effect on cells, while later, lead penetrates only into the root apoplast, and in this way remains harmless to cells.     

Induction of sister chromatid exchanges by heavy metal salts in root meristem cells ofAllium cepa L.

Four heavy metal salts, nickel sulphate, mercuric chloride, cadmium sulphate and zinc sulphate, were tested for induction of sister chromatid exchange (SCE) in root meristem cells ofAllium cepa. A simple modified Feulgen staining procedure was employed for SCE-analysis. Maleic hydrazide and paraquat were included for comparison. An evaluation of genotoxicity of the above test chemicals made on the basis of SCE-assay was found positive for all the test chemicals with exception of zinc sulphate which gave a weak positive result.     

Antifungal saponins from bulbs of white onion,Allium cepa L.

Three saponins, named ceposide A, ceposide B, and ceposide C were isolated from the bulbs of white onion, Allium cepa L. Elucidation of their structure was carried out by comprehensive spectroscopic analyses, including 2D NMR spectroscopy and mass spectrometry, and chemical evidences. The structures of the compounds were identified as (25R)-furost-5(6)-en-1β,3β,22α,26-tetraol 1-O-β-d-xylopyranosyl 26-O-α-d-rhamnoyranosyl-(1 → 2)-O-β-d-galactopyranoside (ceposide A), (25R)-furost-5(6)-en-1β,3β,22α,26-tetraol 1-O-β-d-xylopyranosyl 26-O-α-d-rhamnoyranosyl-(1 → 2)-O-β-d-glucopyranoside (ceposide B), and (25R)-furost-5(6)-en-1β,3β,22α,26-tetraol 1-O-β-d-galactopyranosyl 26-O-α-d-rhamnoyranosyl-(1 → 2)-O-β-d-galactopyranoside (ceposide C). The isolated compounds, alone and in combinations, were evaluated for their antimicrobial activity on ten fungal species. Antifungal activity of all three saponins increased with their concentration and varied with the following rank: ceposide B > ceposide A–ceposide C. We found a significant synergism in the antifungal activity of the three ceposides against Botrytis cinerea and Trichoderma atroviride, because growth of these fungi was strongly inhibited when the three saponins were applied in combination. In contrast, Fusarium oxysporum f. sp. lycopersici, Sclerotium cepivorum and Rhizoctonia solani were very little affected by saponins.     

Effects of successive applications of fosetyl-Al on growth ofAllium cepa L. associated with endomycorrhizal fungi

The effect of successive applications (0 to 4) of fosetyl-Al on the growth of mycorrhizal (M) and nonmycorrhizal (NM) onion plants and on the residual quantity of the fungicide in the root was studied for 6 weeks under growth chamber conditions. Fosetyl-Al was applied as foliar spray at a concentration of 3 mg ml^–1. The successive applications of fosetyl-Al did not affect the dry mass of root or shoot of M plants. NM plants sprayed only once with fosetyl-Al had a higher root mass than plants sprayed 0, 2, 3 and 4 times. The quantity of phosphorous acid, the only residue detected, was positively correlated with the number of fosetyl-Al applications. It showed a positive partial correlation coefficient with the shoot (r=0.41) and the root dry mass (r=0.40) of NM and M plants.Contribution no 344, Station de recherches, Agriculture Canada, Sainte-Foy.     

A contribution to the standardization of methods for the preparation of seed proteins ofAllium cepa L.

The study of certain conditions for the extraction of seed proteins ofAllium cepa revealed that the best extractibility of proteins is obtained by the use of a buffered physiological solution at 20 °C in comparison with TRIS-glycine buffer at 5 °C. Using potassium phosphate buffer with 0.01 M mercaptoethanol and 0.4 M NaCl, an amount of proteins by up to 25 per cent higher passes into solution as compared with the physiological solution, but these extracts are unsuitable for the electrophoretic separation in polyacrylamide gels. The defatting of the seed meal under low temperature did not affect the qualitative composition of the protein complex studied, the addition of a 1 per cent soluble starch to the polyacrylamide gel. improved its resolution.     

A relationship between callose and ectodesmata in epidermal cells of Allium cepa L.

Summary The outer wall of adaxial epidermal cells of onion bulb scales shows punctate callose deposits. The abundance of these callose deposits is highly variable, and seems to correlate with the distribution of ectodesmata. It is proposed that callose is involved in the regulation of ectodesmata as a waterflow regulator and/or molecular filter, thus influencing the cell-environment interaction.Abbreviations LS Linsmaier-Bednar and Skoog - 2,4-D 2,4-ichlorophenoxy-acetic acid - HRGP Hydroxy-proline rich glycoproteins     

Chromosomal variations in the callus tissues ofAllium tuberosum andA. cepa

Summary Chromosome studies ofAllium tuberosum andA. cepa were made from one month to eighteen months old calluses. Different types of chromosomal variations like aneuploid number ranging from 28 to 31, tripolarity, lagging, micronuclei, haploid number etc. were noted inA. tuberosum, whereas inA. cepa the cells showed high chromosome numbers such as 32, 64 or more. The normal chromosome number (2n=16) occurred rarely. The selective pressure of the culture media may have led to the manifestation of the genetic control of differential response to chromosome behaviour and growth in the two species of the same genus.     

Differentiation of metaxylem cell line in the root ofAllium cepa

Summary The first step of differentiation in the root segments ofAllium cepa containing metaxylem cells in different stages of differentiation were studied by DNA reassociation curves and compared to meristem cell extracted DNA. Upon sonication of DNA samples to about 400 base pairs, the reassociation profiles of the heat denatured DNA, were spectrophotometrically followed at two different concentrations. The kinetic complexities,i.e., the number of base pairs per haploid genome of a given sequence and its redundancy were calculated. Differences were found at the level of highly and medium repetitive sequences, thus demonstrating that some DNA reassociation classes may undergo amplification during root development.     

Differentiation of metaxylem cell line in the root ofAllium cepa

Summary DNA was extracted from three root segments ofAllium cepa: i) an apical portion 500 m long from the tip (meristem); ii) a second portion 4 mm long (I root segment containing metaxylem cells in the initial stages of differentiation); iii) a third portion 6 mm long (II root segment containing metaxylem cells in further stages of differentiation). A mixture of homologous 18 S and 25 S³H-rRNA was used for invitro DNA-rRNA hybridization. The following percent saturation values were detected in the three samples: 0.08 in meristem DNA (samplea), 0.129 in I root segment DNA (sampleb), and 0.105 in II root segment DNA (samplec).Thermal denaturation of DNA and the derivative curves of the melting profiles evidenced five DNA families which were differently represented in the three DNA samples. DNA elution by thermal chromatography on hydroxyapatite followed by hybridization with³H-rRNA, revealed that ribosomal cistrons melt between 90 and 91 °C, corresponding to a G-C content of 50.7%. Moreover, the amount of the DNA family containing ribosomal cistrons was greater in sampleb andc, in sampleb to a greater extent, as compared with samplea. On the other hand, one DNA family melting at a higher temperature (92–93 °C) was drastically increased in samplec.Buoyant density profiles of unsonicated DNA showed no peaks in the three DNA samples. Upon somcation, a heavy shoulder was observed in the profile of sampleb. As the density of ribosomal cistrons and that of shoulder were very similar, it seems possible that the two fractions contain many DNA sequences in common.The present studies demonstrate that the proportion of ribosomal cistrons and other DNA families does not keep constant during the development of the metaxylem cell line.     

Effect of trifluralin on the histology and respiration ofAllium cepa root tips

Trifluralin was found to induce isodiametric enlargement ofAllium cepa root tip cells in the elongation zone. It enhanced the endogenous oxygen uptake of the excised root tips. The herbicide was found to have no marked effect on the dry weight of the root tips. It is suggested that the herbicide may exert its effect onAllium cepa root tips through its enhancement of ethylene production.     

Restoration of mitotic and differentiation processes in the root apices ofAllium cepa L. treated with cyanein and griseofulvin

The roots ofAllium cepa L. were treated with water solutions of cyanein and griseofulvin for 24 and 48 h respectively, thereafter cultured for 48 h in the medium without the antibiotics, and the reversibility of the inhibition caused by the antibiotics was evaluated. Changes in the mitotic index of meristematic cells were followed in squash preparations of root apices. In addition to the cytologic observations, the differentiation of the primary meristem was followed in longitudinal sections. After the treatment with reversible doses of cyanein and griseofulvin respectively, differences were found in both the effects of the antibiotics during the treatment and the restoration of the inhibited processes after the treatment. In the cells treated with cyanein the restoration of mitotic activity was instantaneous but less intensive and in|complete especially in the case of the 48 h treatment. After the treatment with griseofulvin which temporarily interfered also with the differentiation of the primary meristem, the restoration of the inhibited processes was delayed but intensive after both application times, 48 h after the treatment no symptoms of the preceding inhibition could be observed.