Meristematic Cells Of Vicia Faba Research Articles

Cyto 2011—Overview of the XXVI ISAC Congress Proceedings Issue of Cytometry

THIS issue of Cytometry presents the third occasion on which a call has been issued to encourage authors to submit their work to the journal for publication as part of a special Congress Proceedings Issue of Cytometry. An incentive for submission was that accepted papers would be guaranteed an oral presentation at the XXVI ISAC Congress in Baltimore—CYTO 2011. The topics of the accepted papers range from proteomewide modeling to CD4 testing in resource-limited settings. The research articles do not speak to a given theme, rather they provide a snap-shot of the diversity of interest in cytometry with proffered examples of some of the latest trends in the field of cytometry. The Proceedings Issue of Cytometry also incorporates two reviews. The first review, by Darzynkiewicz et al., has been scheduled for publication in alignment with the conference, as it speaks to a topical area—‘‘click chemistry’’—an increasingly used conjugation and assembly technology with attendant advantages of biocompatibility, selectivity, and yield (1). The second review, presented at CYTO 2011, provides an update on cytometric challenges in veterinary andrology. In parallel with the discovery of the structure of DNA, 1953 also saw the publication of a study by Howard and Pelc (2) on the patterns of radiolabel incorporation by meristematic cells of Vicia faba. This landmark paper proposed the existence of a cell division cycle demarcated by preand postgaps for DNA synthesis. Their discovery set in process the widespread application of labeling methods for the analysis of the eukaryotic cell cycle, leading to the recognition of the universality of its major regulators (3) and exploitation in studies on anticancer drug-induced perturbations of cell cycle kinetics. In this issue, Darzynkiewicz et al. overview the Cytometry of DNA replication and RNA synthesis from its historical backdrop to recent advances based on ‘‘Click Chemistry.’’ The review covers fluorescence quenching methods, immunocytochemical detection through to the recent incorporation of EdU for the click chemistry detection of DNA damage via histone H2AX phosphorylation analyzed by flow or laser scanning cytometry. K.B. Sharpless, the 2001 Nobel Prize winner for Chemistry, introduced the concept of click chemistry for reactions that fulfill useful criteria including being modular and providing very high yields with favored physiologically stable, single reaction products (4). The concept drives applications for addition reaction conditions using preferred solvents and simple routes for product isolation— key issues for the cytometrist. Since the first reports of sexed sperm, flow technology efforts have been largely focused on improving sample throughput resulting in routine high-purity sorting of Xor Y-chromosome-bearing sperm (5). However, a more refined assessment of fertilizing potential of veterinary semen samples by flow cytometry has yet to exploit the full potential of the technology. Petrunkina and Harrison in Cambridge, UK have worked on the determinants of sperm quality and fertility in domestic species and recently provided an exact mathematical analysis of the systematic mis-estimation of semen-derived cell subpopulations by flow cytometry (6). The current review addresses cytometric solutions presenting recent developments, advantages and limitations together with the aspects of sperm physiology that could benefit from multicolor flow cytometric procedures. Advantageous biophotonic characteristics of molecular probes underpin many cytometric studies. The detection of lu-

Red cabbage anthocyanin extract alleviates copper-induced cytological disturbances in plant meristematic tissue and human lymphocytes

Red cabbage is a source of health beneficial substances with antioxidant and antigenotoxic properties. HPLC analysis specifying the content of the investigated extract indicated that mainly anthocyanins (ATH) were responsible for its abilities. Cytological research was conducted with two experimental models: plant tissues--meristematic cells of Vicia faba, and animal tissue elements--human lymphocytes. Positive influence of ATH extract on mitotic activity of Vicia cells exposed to Cu(2+) stress, and inhibitory effect of ATH on cytotoxic actions of Cu(2+) on lymphocytes were demonstrated. In all experimental series with ATH application in combinations with Cu(2+), mitotic index (MI) were higher than those obtained for only Cu(2+) stressed tissues. Preincubation in ATH before Cu(2+) stress had the best effect. Similarly, after ATH applications in all tested series decrease in frequency of micronuclei (MN) appearance was noticed in comparison with only Cu(2+) stressed material. In the case of Vicia cells ATH acted effectively even applied after Cu(2+) stress. It suggests that this ATH mixture not only prevents and limits but also heals the cytological injury caused by Cu(2+) stress.

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

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

The effect of continuous gamma irradiation on formation of sister chromatid exchanges (SCEs) and chromosomal aberrations in meristematic cells ofVicia faba

Germinated seeds ofVicia faba were continuously irradiated at low dose rate of gamma rays (0.05 Gy h-1) up to a total accumulated dose of 2 Gy. The FPG (fluorescence plus Giemsa) technique of differential chromatid staining was used to monitor the frequency of sister chromatid exchanges (SCEs) in irradiated root tip meristem cells. The results of the experiments have demonstrated that SCE frequency is raised by continuous gamma irradiation only in plant cells containing BrdU in the chromosomal DNA. No effect concerning SCE formation was recorded at continuous irradiation of meristematic cells of Vicia faba with native, i. e. BrdU-nonsubstituted, DNA. In contrast to SCEs, a significant increase was found in the yield of chromosomal aberrations in all variants of irradiation.

Chromatin-free compartments and their contents in anaphase chromosomes of higher plants

Chromatin-free compartments and their contents were observed in chromosomes of root tip meristematic cells of Vicia faba and Allium cepa . The central chromatin-free compartment of chromosome was usually larger than the peripheral ones. In tranverse section, every chromosome showed peripheral chromatin-free compartments though their number and size varied. In longitudinal section, the central chro matin-free compartments were found to be distributed intermittently along the chromosomal axis. The contents, which showed a granular-fibrillar nature and linked up to the rest of the chromosome, were seen in the central and at least some of the peripheral chromatin-free compartments. A cytochemical study revealed that these contents consisted of ribonucleoprotein granules and fibrils. The contents are a special constituent of the chromosome. The chro matin-free compartments in a chromosome seemed connected with one another and their contents may form a network system which plays a role in the maintenance and stabilization of the higher-order structure of chromosome.

Origin of nucleolus-like bodies found in the nucleoplasm and cytoplasm of Vicia faba meristematic cells

Cytohistochemical staining and RNase-gold labelling have been applied to root-tip meristematic cells of Vicia faba to study the origin and biological significance of 2 types of inclusions: one seen in the nucleoplasm and the other in the cytoplasm of early telophase cells. They have been termed “dense bodies” and “cytoplasmic nucleolus-like bodies” (NLB), respectively. Both types of inclusions respond positively to silver staining and ribonucleoprotein (RNP) staining in a similar fashion to nucleolus. Interestingly, the dense bodies label heavily with the RNase-gold complex, as does the nucleolus, while the cytoplasmic NLB have no affinity with the label. In most cases, the dense bodies label more heavily than the nucleolus. Light microscope surveys reveal that the dense bodies sometimes appear to be released from the surface of the nucleolus. On the other hand, prenucleolar material showing the same silver staining and RNP preferential staining characteristics as the dense bodies begin to accumulate on the surface of chromosomes in mid-anaphase. This material does not label with RNase-gold. These data are discussed in terms of the hypothesis that the dense bodies are derived from the nucleolus by direct budding or fragmentation, and the cytoplasmic NLB are composed of prenucleolar material that failed to attach to chromosomes.

A Scanning Electron Microscopic Observation on the Nucleolus in the Root Meristematic Cells of <italic>Vicia faba</italic> (Broad Beans)

Journal Article A Scanning Electron Microscopic Observation on the Nucleolus in the Root Meristematic Cells of Vicia faba (Broad Beans) Get access Keizo MARUYAMA, Keizo MARUYAMA Biological Laboratory, Yoshida College, Kyoto UniversitySakyo-ku, Kyoto, 606 Japan Search for other works by this author on: Oxford Academic PubMed Google Scholar Mitsuro OKUDA Mitsuro OKUDA Biological Laboratory, Yoshida College, Kyoto UniversitySakyo-ku, Kyoto, 606 Japan Search for other works by this author on: Oxford Academic PubMed Google Scholar Journal of Electron Microscopy, Volume 31, Issue 3, 1982, Pages 253–256, https://doi.org/10.1093/oxfordjournals.jmicro.a050365 Published: 01 January 1982 Article history Published: 01 January 1982 Received: 28 January 1982 Accepted: 31 May 1982

Účast exogenní DNK v reparačních procesech meristematických buněkVicia faba poškozených monofunkěním alkylaěním ěinidlem ethylmethan sulfonátem

The effect of exogenous DNA of syngeneic origin on the course of reparation of meristematic cells ofVicia faba primary roots followed after treatment with monofunctional alkylating agent, ethyl methanesulphonate (EMS), was tested. Time course of alternations in mitotic activity of investigated cellular population and the dynamics of formation of postmetaphase chromosomal aberrations was evaluated. A reparation of damaged cells was significantly supported by syngeneic DNA; its application induced an increased incidence of cellular division already in the early intervals of the repair which was accompanied by concomitant decrease of microscopically detectable rupture in the chromosomes. The study performed on the localization of induced damages occurring in metaphase chromosomes pointed out an increased sensitivity of small chromosomes ofVicia faba to EMS. Similarly, a reparative action of syngeneic DNA was exhibited by significant decrease of aberrations frequency, predominantly in the same chromosomal group. Per cent representation of individual types, not affected by the action of syngeneic DNA, was established by detailed classification of induced aberrations. In both cases, isochromatide breaks were found of greatest predominance.

The effect of adenosine triphosphate on mitomycin C-induced aberration yield in Vicia faba.

The effect of adenosine triphosphate (ATP) on the yield of mitomycin C-induced chromosome aberrations was examined with the root meristematic cells of Vicia faba. ATP content of root tips, as determined by luciferin-luciferinase assay, increased by over twofold after treatment in an ATP solution. The results indicated that ATP, given as a pre- or post-treatment, reduced the aberration yield. The equal efficacy of both pre- and post-ATP treatment in the modification of aberration yield may suggest a significant biochemical role played by ATP during the recovery period. The suggestion is further made that ATP serves as an energy source for a metabolic process which regulates repair of damaged chromosomes.

The effect of BUdR on the cycle of irradiated and unirradiated meristematic cells of Vicia faba

The cell cycle time of the meristematic cells of Vicia faba was approximately 20 hr in both the control cells and in the cells treated with TdR (1·6 × 10−5 m ) for 24 hr. Cells treated for 24 hr with an equimolar concentration of BUdR showed prolongation of their cell cycle time to about 24 hr. The main detectable change in the cell cycle was the lengthening of the S phase which was increased to 13 hr from 8 hr which was the value for the S phase of the control and TdR-treated cells. BUdR-treated cells exhibited more radiation-induced damage, measured as micronuclei, than control or TdR-treated cells. The radiosensitization effect was manifested as an increase in the percent of damaged cells but not as an increase in the damage per damaged cell. An attempt to correlate the BUdR-induced changes in the cell cycle with the BUdR-induced increase in radiation response was presented.

The effect of maleic hydrazide and exogenous DNA on chromosomes ofVicia faba

The mitotic activity of merstematic cells ofVicia faba, the frequency of chromosomal abnormalities and their interchromosomal distribution are evaluated in dependence on the concentration of maleic hydrazide (MH) and on the recovery period. The influence of exogenous DNA of different genetic origin on the course of repair of primary root cells damaged by MH was also studied. Isologous DNA which exhibited a strong repair effect in authors' previous experiments was quite ineffective in the case of maleic hydrazide. Heterologous DNA, on the other hand, had to some extent a parallel effect with MH in breaking down the structural integrity of chromosomes and increased the frequency of aberrations induced by maleic hydrazide in meristematic cells ofVicia faba.

Contribution to the study on the reparative effect of exogenous DNA in the irradiated meristem ofVicia faba

The present report is focused on the study of participation of exogenous DNA in the process of postirradiation reparation of meristematic cells ofVicia faba primary roots. It is aimed at comparison of the positive reparative effect of isologous DNA with postirradiation action of heterologous DNA in its native, thermally denatured and DNAase-degraded forms, or DNA degraded by ultrasound, and with the effect of other biologically important macromolecules (RNA, histone, heparin, and dextran sulphate). For this purpose, the roots ofVicia faba irradiated by 150 r exposure were cultivated in solutions containing the above substances for an appropriate time interval. In squash slides both mitotic activity of the investigated cell population and frequency of postmetaphase chromosomal aberrations induced by radiation were evaluated. It was shown that a stimulation of cell division and reparation of chromosome damages were supported exclusively by isologous DNA. On the contrary, exogenously applied heterologous DNA increased postirradiation frequency of aberrations; maintenance of native structure of applied DNA was an essential condition for the above effect. Other macromolecules investigated on the course of postirradiation reparation ofVicia faba meristematic cells were without effect.

Nativita makromolekuly isologní DNK jako podmínka jejího reparačního efektu při radiačním poškození

The effect of isologous DNA on the course of postirradiation reparation of meristematic cells ofVicia faba primary roots was studied in detail. A considerable interest was devoted to determinations of fundamental qualitative and quantitative conditions of the above effect of isologous DNA. Main criteria of the effect were both mitotic activity of irradiated cellular population and dynamics of chromosome aberrations induced by radiation. One set of experiments compared the course of reparation as occurred in regard to applied dose of ionizing radiation in native isologous DNA, DNA denaturated by heat and degraded by DNAase, and post-irradiation reparation of induced damages was favorably affected by native isologous DNA only. Another set of results evaluated the dependence of positive reparative effect of native isologous DNA on the length of the molecule demonstrating that in the process of reparation the presence of a complete DNA macromolecule was not essential. The last experimental group was focused on observations on the dependence of the rate of native isologous DNA effect on concentration of applied solution of the macromolecule.

Nativity of the macromolecule of isologous DNA as a condition for its reparative effect on radiation damage

The effect of isologous DNA on the course of postirradiation reparation of meristematic cells ofVicia faba primary roots was studied in detail. A considerable interest was devoted to determinations of fundamental qualitative and quantitative conditions of the above effect of isologous DNA. Main criteria of the effect were both mitotic activity of irradiated cellular population and dynamics of chromosome aberrations induced by radiation. One set of experiments compared the course of reparation as occurred in regard to applied dose of ionizing radiation in native isologous DNA, DNA denaturated by heat and degraded by DNAase, and post-irradiation reparation of induced damages was favorably affected by native isologous DNA only. Another set of results evaluated the dependence of positive reparative effect of native isologous DNA on the length of the molecule demonstrating that in the process of reparation the presence of a complete DNA macromolecule was not essential. The last experimental group was focused on observations on the dependence of the rate of native isologous DNA effect on concentration of applied solution of the macromolecule.

Contribution to the study on the reparative effect of exogenous DNA in the irradiated meristem ofVicia faba

The present report is focused on the study of participation of exogenous DNA in the process of postirradiation reparation of meristematic cells ofVicia faba primary roots. It is aimed at comparison of the positive reparative effect of isologous DNA with postirradiation action of heterologous DNA in its native, thermally denatured and DNAase-degraded forms, or DNA degraded by ultrasound, and with the effect of other biologically important macromolecules (RNA, histone, heparin, and dextran sulphate). For this purpose, the roots ofVicia faba irradiated by 150 r exposure were cultivated in solutions containing the above substances for an appropriate time interval. In squash slides both mitotic activity of the investigated cell population and frequency of postmetaphase chromosomal aberrations induced by radiation were evaluated. It was shown that a stimulation of cell division and reparation of chromosome damages were supported exclusively by isologous DNA. On the contrary, exogenously applied heterologous DNA increased postirradiation frequency of aberrations; maintenance of native structure of applied DNA was an essential condition for the above effect. Other macromolecules investigated on the course of postirradiation reparation ofVicia faba meristematic cells were without effect.

Action of Ypenyl [5-bis-(2-chloroethyl)-aminomethyluracil] on the chromosomes ofVicia faba

An investigation was carried out on ability of Ypenyl,i.e. 5-bis-(2-chloroethyl)-aminomethyluracil to induce structural disturbances of chromosomes and changes of mitotic activity of meristematic cells ofVicia faba. The cytostatic employed induced chromosomal aberrations mostly of the chromatide type, the greater part of which were localized in the group of small chromosomes. The frequency of aberrations was proportional to the concentration of the substance used as well as to the duration of treatment. After a 24 hour treatment with 0·01mm solution of Ypenyl the maximum incidence of aberrations was found after 8 hour recovery in water. Longer recovery periods resulted in a gradual decrease in the number of aberrant post-metaphases in the cell population studied. Increasing concentration of Ypenyl led to the decrease in mitotic activity. Short recovery periods caused a considerable decrease in the mitotic index, which was compensated as late as 16 hours after treatment with the cytostatic.

Влияние извне поданной ДНК разного происхождения на митоз и хромосомы облученных меристематических клеток Vicia 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.

Metabolic DNA in heart and skeletal muscle and in the intestine of mice.

STUDIES of the incorporation of labelled thymidine into the DNA of heart muscle and skeletal muscle in adult animals have shown significant values which indicate regular renewal of some or all of the DNA in these non-dividing organs1,2. Autoradiography has shown labelled muscle nuclei in smooth muscle and in heart muscle and therefore subsidiary cells, such as lymphocytes, cannot be held responsible for the labelling. The existence in plant materials of two separable fractions of DNA with different rates of turnover has been shown by Sampson et al.3, and Sampson and Davies4 have presented good evidence for the location of a fraction (DNA1) with a high rate of turnover in differentiating cells and of a stable fraction (DNA2) in meristematic cells of Vicia faba. The results of experiments designed to compare the specific activities of the DNA fractions with a molecular weight below 4 × 106 with those above 4 × 106 in heart, skeletal muscle and intestine are reported here.

NUCLEOLONEMA AND PARS AMORPHA IN ROOT MERISTEMATIC CELLS OF VICIA FABA

In osmium-fixed, Epon-embedded root meristematic cells of Vicia faba stained according to the Feulgen – methylene blue procedure, the interphase nucleolus is seen to consist of a metachromatic cabbage-green colored material in which are embedded numerous unstained vacuole-like structures of varying size. By using an orange Wratten filter in the illumination system, it is observed that the metachromatically stained nucleolar material is, in fact, made up of two components, each segregated into zones distinguishable by differences in staining intensity. The first more lightly stained component is found in the central and bordering regions of the nucleolus as well as in a number of areas of varying width extending radially in between them. The second more intensely stained component is observed in areas of varying length and width usually occupying the intermediate regions of the nucleolar mass. Under electron microscopy, the first component is seen to consist of densely packed fibrils and granules some 150 Å in diameter; in places, those constituents are assembled into seemingly coarse, thread-like structures approximately 0.1 μ in diameter. The second component, on the other hand, is made up mainly, if not exclusively, of tightly packed fibrils 60 to 100 Å in diameter. Judging from their topographical distribution and ultrastructural characteristics, it would seem as if the two nucleolar components just described correspond respectively to what is usually referred to as nucleolonema and pars amorpha in animal nucleoli. In addition to nucleolonema and pars amorpha, two ultrastructurally and topographically distinct types of vacuole are recognizable within the interphase nucleolus of Vicia faba. The vacuoles of the first type are usually large or medium-sized, contain loosely and uniformly scattered fibrils and granules approximately 150 Å in diameter, and are observed exclusively within the nucleolonematic zones of the nucleolus. The vacuoles of the second type are small, contain only loosely dispersed fibrils, 60 to 100 Å in diameter, and are confined to the pars amorpha of the nucleolus.

EVIDENCE FOR THE EXISTENCE OF TWO TYPES OF NUCLEOLAR RIBONUCLEOPROTEIN IN ROOT MERISTEMATIC CELLS OF VICIA FABA

In Helly-fixed root meristematic cells of Viciafaba, the toluidine blue – molybdate staining procedure reveals the existence within the interphase nucleus of two metachromatically distinct types of nucleolar ribonucleoprotein. The ribonucleoprotein of the first type stains bluish-green and is associated with the nucleolar matrix. The ribonucleoprotein of the second type takes on a bright purple color and is present inside the nucleolar vacuoles. Evidence is presented suggesting that, during interphase, the nucleolus is the site of active synthesis and of accumulation inside its vacuoles of the second type of ribonucleoprotein.