Effect of osmotic pressure on root growth,cell cycle and cell elongation

Summary The paper reports a study of root growth, the duration of the cell division cycle and cell size, inAllium cepa roots grown in mannitol solution, with osmotic pressures of 0–16 atm., at 25° C, with aeration. Root growth decreases markedly as the osmotic pressure rises, at 12 atm. being about 35% of what it is at 0 atm. The rate of the cell cycle, , expressed as the percentage of cells passing through any given point in the cycle in one hour, is a roughly linear function of the osmotic pressure, and at 12 atm. is reduced to 80% of what it is at 0 atm. The reaction of the cell size to osmotic pressure is similar to that of the growth. The relative values of the two diverge progressively as the osmotic pressure increases and at 12 atm. the elongation of the cells has dropped to 40% of normal.The data obtained agree with those given by the equationG = K · · L, in whichK is a constant, is the rate of the cycle (reciprocal of its duration) andL the average size of the mature epidermal cells.     

Growth components in Allium roots

Summary This paper reports results of a study of root growth, duration of the division cycle, and cell size for different intervals of root length, associated with different phases of growth, namely, from the onset of growth to the achievement of a steady state characterized by a constant rate of growth. The material is roots of Allium cepa.Steady-state kinetics of growth are achieved only when all components of growth reach a constant value. Although the cell flow remains constant from a root length of 8 mm onwards, the steady condition is only observed when the final size of the epidermal cells reaches its maximum and constant value (at about 12 mm of root length).The equation G=24×10^-5 \N\·L is proposed to analyze the growth of Allium roots in its components. G is the growth rate (mm/24 hr), N the number of cells per column in the meristematic ribs, the rate of the cell cycle (the reciprocal of its duration x100), and L the average final size of the epidermal cells (in ).     

Inhibition of bacterial segregation by early functions of phage Mu and association of replication protein B with the inner cell membrane

Summary Infection of Mu-sensitive bacteria with a recombinant phage that carries the EcoRI·C fragment from the immunity end of wild type Mu DNA causes filamentous growth. Transmission electron microscopy revealed that the cell-division cycle was inhibited at, or prior to, the initiation of septation. The filamentation does not occur after infection of Mu-immune bacteria or after infection with a phage carrying the same EcoRI·C fragment, but with an IS1 insertion in gene B of Mu, showing that either gpB and/or some non-essential functions (e.g. kil) mapping downstream from the insertion are required for the inhibition of cell division. These data and previously published evidence suggest that in the killing of E. coli K12 by early Mu functions expressed from the cloned EcoRI·C fragment, two components have to be distinguished: one, a highly efficient elimination of plasmid DNA carrying the early Mu genes, and second, a series of interactions with host functions conducent to an inhibition of cell division. It is suggested that functions normally involved in the SOS reaction participtate in the inhibition of cell division by early Mu functions. Infected bacteria synthesize the replication protein B (M_R 33000) of Mu, which was found by cell fractionation experiments to be associated with the inner cell membrane. The role of this association for filamentous growth and for the integrative replication of the phage is discussed. The recombinant phage might be useful as a tool for the study of the E. coli cell division cycle.     

Synchronized cultures of a cell wall-less mutant of Chlamydomonas reinhardii

Synchronization and synchronous growth of a cell wall-less mutant of Chlamydomonas reinhardii have been described. The following growth conditions were used: A modified Sueokas' high salt minimal medium, 1410 h light-dark cycle, growth temperature 30°C, light intensity 12–18 Klux and dilution of the culture at the end of the dark to a constant cell density of 1.0·10⁶ cells/ml. The time course of increase and distribution of cell volume, cytoplasmic and nuclear division, release of motile cells after the division period and accumulation of DNA, RNA and protein are reported. These mutant cells did not make any sporangium in which the dividing cells were kept as a unit inside a mother cell wall. However, they usually adhered during the period of division, thus making clumps containing 2, 4 and 8 cells. Several of these cell clumps dissolved releasing either single or couples of 2 and 4 cells. After the end of division the cells became flagellated and motile and thereby releasing themselves from the aggregate.Non-Standard Abbreviations AWV average weighed cell volume - MM minimal medium - HSM high salt medium - TCA trichloroacetic acid     

A temperature sensitive mutation that reduces mitotic rate inDrosophila melanogaster

Summary A temperature-sensitive cell autonomous mutation ofDrosophila, l(1)ts-1126 (1–16±2), that affects the rate of cell division is described. When mutant animals are exposed to the restrictive temperature of 29°C during the first and second larval stages, the growth rate of the larvae is retarded. A delay in pupariation occurs during which larvae reach their full size, and the resulting flies are normal. When mutant animals are exposed to restrictive temperature during the third larval stage, growth is also retarded but no delay in pupariation occurs, and the resulting flies are reduced in size. Their small size is due in part to a decreased number of cells and in part to a smaller size of the cells.X-ray induced, marked, homozygousl(1)ts-1126 clones in an otherwise normal animal, are smaller in animals exposed to pulses of restrictive temperature when compared to clones in animals kept at permissive temperature of 22°C. Clone size decreases as pulse length increases. Clones on the wing blade induced 24 h after oviposition are smaller than clones induced at 48 h in animals grown at restrictive temperature. This result is interpreted as an inability of the slower dividingl(1)ts-1126 cells to survive when in competition with wildtype cells. The distribution of survivingl(1)ts-1126 clones in gynandromorphs grown at restrictive temperature supports this conclusion.     

Plant tyrosine decarboxylase can be strongly inhibited by l-α-aminooxy-β-phenylpropionate

The effect of tunicamycin, an inhibitor of N-glycosylation of proteins, on growth and on synthesis of DNA and protein was studied in suspension cultures from Nicotiana tabacum and Catharanthus rosea. In the presence of 0.1–1 g · ml^-1 tunicamycin, cell division and DNA synthesis stopped in cells which had been proliferating logarithmically, but protein formation continued. Cytophotometric determination of the nuclear DNA content in Catharanthus cells showed that a cell-cycle arrest had occurred in G1 phase. Metabolic labelling of cells with the glycoprotein precursors glucosamine or mannose was inhibited, too. The results indicate that one or more glycoproteins are needed for the cell to pass through the G1 phase, as was recently postulated for animal and yeast cells.Abbreviations TCA trichloroacetic acid - TM tunicamycin     

Control of cell size at division in fission yeast by a growth-modulated size control over nuclear division

In the fission yeast Schizosaccharomyces pombe, nutritional reduction of growth rate by supplying poor nitrogen, carbon or phosphate sources causes a decrease in cell size. The effect on cell division following three different nutritional shifts-up has been investigated. In all cases, about 20% of the cells divide at the original cell length, and then cell division stops for a period. Cell division then resumes at the new faster rate, cell length at division being characteristic of the new medium. Further investigation reveals that the first effect of the shift is to inhibit nuclear division rapidly and completely. These results are strongly suggestive of the operation of a cell size requirement for entry into nuclear division. The cell size necessary for nuclear division is set, or modulated, by the prevailing growth conditions. This model is confirmed by a nutritional shift-down, where nuclear division and cell division are stimulated after the shift. Cell length at division falls rapidly until the new shorter length is attained, when a new steady state is assumed at a slower growth rate. The control system is compared with that in bacteria, and its implications for various models proposed for the control of timing of mitosis are discussed.     

Callus production from willow (Salix viminalis L.) protoplasts

Protoplasts were isolated from cell suspensions of Salix viminalis (basket willow) clone 78-0-90 and S. schwerinii clone 77-0-77, using cellulysin and macerase in modified Woody Plant medium. For clone 78-0-90, 6.3 · 10⁶ ± 1.9 · 10⁶ protoplasts were obtained per gram fresh weight. Cell divisions started two days after protoplast isolation and gave rise to callus which has been maintained in culture for up to four years. Protoplast yield from the clone 77-0-77 was lower (less than 10⁶ protoplasts per gram cells), cell division was infrequent and no callus was obtained. Protoplasts were also isolated from the leaves of willow shoot cultures using cellulysin and pectolyase, but these did not show cell divisions.Abbreviations BA benzyladenine - 2,4-D 2,4-dichlorophenoxyacetic acid - MS medium Murashige & Skoog (1962) medium - WP medium Woody Plant medium (Lloyd & McCown 1981)     

A Generalised Age- and Phase-Structured Model of Human Tumour Cell Populations Both Unperturbed and Exposed to a Range of Cancer Therapies

We develop a general mathematical model for a population of cells differentiated by their position within the cell division cycle. A system of partial differential equations governs the kinetics of cell densities in certain phases of the cell division cycle dependent on time t (hours) and an age-like variable τ (hours) describing the time since arrival in a particular phase of the cell division cycle. Transition rate functions control the transfer of cells between phases. We first obtain a theoretical solution on the infinite domain −∞ < t < ∞. We then assume that age distributions at time t=0 are known and write our solution in terms of these age distributions on t=0. In practice, of course, these age distributions are unknown. All is not lost, however, because a cell line before treatment usually lies in a state of asynchronous balanced growth where the proportion of cells in each phase of the cell cycle remain constant. We assume that an unperturbed cell line has four distinct phases and that the rate of transition between phases is constant within a short period of observation (‘short’ relative to the whole history of the tumour growth) and we show that under certain conditions, this is equivalent to exponential growth or decline. We can then gain expressions for the age distributions. So, in short, our approach is to assume that we have an unperturbed cell line on t ≤ 0, and then, at t=0 the cell line is exposed to cancer therapy. This corresponds to a change in the transition rate functions and perhaps incorporation of additional phases of the cell cycle. We discuss a number of these cancer therapies and applications of the model.     

Uncoupling growth from the cell cycle

How does a Drosophila wing grow to the appropriate size and shape? Although a collaboration of cell division with the patterning of cell fates seems obvious, almost nothing is known about how these two processes are coordinated during development. A recent paper¹ finds that blocking cell division uncouples cell growth from the cell division cycle, displaying remarkable flexibility in the ability of the wing primordia to achieve the right proportions with fewer than normal cells. BioEssays 20: 283-286, 1998.© 1998 John Wiley & Sons, Inc.     

Growth characteristics of flagellated cells of Phaeocystis pouchetii revealed by diel changes in cellular DNA content

The present study reports on effects of different light:dark periods, light intensities, N:P ratios and temperature on the specific growth rate of flagellated cells of Phaeocystis pouchetii in culture. The specific growth rate was estimated by diel changes in cellular DNA content. The cellular DNA content and cell cycle of flagellated cells of P. pouchetii are shown, and the importance of light:dark period in cell division is demonstrated. Diel patterns of the cellular DNA content showed that cell division was confined to the dark period. The cells dealt with more than one division per day by rapid divisions shortly after each other.The specific growth rates (μ_DNA) based on the DNA cell cycle model were in close agreement with specific growth rates (μ_Cell) determined from cell counts. The temperature affected the specific growth rates (multiple regression, p < 0.01) and were higher at 5 °C (μ ≤ 2.2 d⁻¹) than at 10 °C (μ ≤1.6 d⁻¹). Increasing the light:dark period from 12:12 h to 20:4 h affected the specific growth rate of P. pouchetii at the lower temperature tested (5 °C) (multiple regression, p < 0.01), resulting in higher specific growth rates than at 10 °C. At 10 °C, the effect of light:dark period was severely reduced. Neither light nor nutrients could compensate the reduction in specific growth rates caused by elevated temperature. The specific growth rates was not affected by the N:P ratios tested (multiple regression, p = 0.21). The experiments strongly suggest that the flagellated cells have a great growth potential and could play a dominating role in northern areas at increased day length.     

Estimation of parameters in population models for Schizosaccharomyces pombe from chemostat data

The integro-differential growth model of Eakman, Fredriekson, and Tsuehiya has been employed to fit cell size distribution data for Schizosaccharomyces pombe grown in a chemostat under severe product inhibition by ethanol. The distributions were obtained with a Coulter aperture and an electronic system patterned after that of Harvey and Marr. Four parameters—mean cell division size, cell division size standard deviation, daughter cell size standard deviation, and a growth rate coefficient—were calculated for models where the cell growth rate was inversely proportional to size, constant, and proportional to size. A fourth model, one where sigmoidal growth behavior was simulated by two linear growth segments, was also investigated. Linear and sigmoidal models fit the distribution data best. While the mean cell division size remained relatively constant at all growth rates, standard deviation of division size distribution increased with increasing holding times. Standard deviation of the daughter size distribution remained small at all dilution rates. Unlike previous findings with other organisms, the average cell size of Schizosaccharomyces pobme increased at low growth rates.     

The effect of carbon dioxide on the growth kinetics of fructose-limited chemostat cultures of Acetobacterium woodii DSM 1030

The growth of the anaerobic acetogenic bacterium Acetobacterium woodii DSM 1030 was investigated in fructose-limited chemostat cultures. A defined medium was developed which contained fructose, mineral salts, cysteine · HCl and Ca pantothenate (1 mg · 1^–1) supplied in a vitamin supplement. Growth at high dilution rates was dependent on the presence of CO₂ in the gas phase. The ^max was found to be 0.16 h^–1 and the fructose maintenance requirement was 0.1 to 0.13 mmol fructose · (g dry wt)^–1 · h^–1. A growth yield of 61 g dry wt · (mol fructose)^–1, corrected for the cell maintenance requirement and for incorporation of fructose carbon into cell biomass, was determined from the fructose consumption. A corresponding growth yield of 69 g dry wt · (mol fructose)^–1 was calculated from the acetate production assuming that fructose fermentation was homoacetogenic. A Y_ATP of 12.2 to 13.8 g dry wt · (mol ATP)^–1 was calculated from these growth yields using a value of 5 mol ATP · (mol fructose)^–1 as an estimate of the amount of ATP synthesised from fructose fermentation. The addition of yeast extract (0.5 g · 1^–1) to the medium did not influence the ^max or cell yield. After prolonged growth under fructose-limited conditions the requirement of the culture for CO₂ in the gas phase was reduced.Abbreviations YE yeast extract - IC inorganic carbon - D fermenter dilution rate : h^–1 - M_X maintenance requirement for X: mmol X · (g dry wt)^–1 · h^–1 - X may be fructose (Fruct), fructose consumed in energy metabolism (Fruct [E]), acetate (Ac) - ATP CO₂, NH _inf4 ^sup+ or Pi - qX specific rate of utilisation or consumption of X: mmol X · (g dry wt)^–1 · h^–1 - V fermenter volume: litre - rC · Cell, fermenter cell carbon production: mmol C · h^–1 - Y_X yield of cells on X: g dry wt · (mol X)^–1 - Y _infx ^supmax the yield corrected for cell maintenance: g dry wt · (mol X)^–1 - S_ATP stoichiometry of ATP synthesis from fructose: mol ATP · (mol frucose)^–1 - x cell concentration: g dry wt · 1^–1 - specific growth rate : h^–1 - ^max maximum specific growth rate: h^–1     

Suspension feeding in Bithynia tentaculata (Prosobranchia,Bithyniidae), as affected by body size,food and temperature

The suspension feeding of Bithynia tentaculata was tested in laboratory experiments. The animals were fed in 1-1 aerated glass beakers, and filtration rates were calculated from changes in cell concentrations during the 6-h experiment. Temperature influenced the filtering rate, with minimum values of 5ml · ind^–1 · h^–1 at 5° C and maxima of 17.2 ml · ind^–1 · h^–1 at 18° C. Three food species of different size, motility and cell surface characteristics (Chlamydomonas reinhardii, Chlorella vulgaris and Chlorogonium elongatum) did not affect filtration rates. Suspension feeding increased with increasing food concentrations up to 12 nl · ml^–1, above which feeding rate was kept constant by lowering the filtering rates. Even the smallest animals tested (<4 mm body length) were found to be feeding on suspended food at a rate of 2.7 ml · ind^–1 · h^–1, and increasing rates up to 8.4 ml were found in the 6–7 mm size class. All size classes of Bithynia showed a circannual fluctuation of their filtration rates. The ecological consequences of Bithynia's ability to switch between two feeding modes, grazing and suspension feeding, are discussed.     

Growth and productivity of the psychrophilic marine diatoms Thalassiosira antarctica Comber and Nitzschia frigida Grunow in batch cultures at temperatures below the freezing point of sea water

Summary The diatoms Nitzschia frigida and Thalassiosira antarctica grow exponentially even at temperatures between-4 and -6°C and a salinity between 73 and 100 Under these conditions the light saturation of growth is reached in continuous light at a scalar quantum irradiance of between 7 mol·m^–2·s^–1 and 10 mol · m^–2 · s^–1. The increase in salinity retards growth more than a decrease in temperature. For N. frigida the limit of growth is at -8°C (S = 145%.). At increasing quantum irradiance, the chlorophyll content per unit cell volume decreases, whereas there is a significant increase in the carbon content of the exponentially growing cells. In addition, there is hardly any change in the protein content. The results show that both species of diatom can survive in ice without forming resting spores and even grow at extremely low temperatures.     

Living cell reaction process forl-isoleucine andl-valine production

Summary A new process (Living Cell Reaction Process) forl-isoleucine production using viable, non-growing cells ofBrevibacterium flavum AB-07 was optimised using ethanol as the energy source and -ketobutyric acid (-KB) as precursor.l-valine also could be produced from glucose at high yield by this process. This process differs from the usual fermentation method in that non-growing cells are used, and the production ofl-isoleucine andl-valine were carried out under conditions of repressed cell division and growth. Minimal medium missing the essential growth factor, biotin was employed as the reaction mixture for the production ofl-isoleucine andl-valine. The productivity ofl-isoleucine andl-valine were 200 mmol·l^–1 · day^–1 (molecular yield to -KB: 95%) and 300 mmol · l^–1 · day^–1 (molecular yield to glucose: 80%) respectively. The content ofl-isoleucine andl-valine in total amino acids produced in the each mixture were 97% and 96% respectively.     

Modulating zucchini cotyledon plate meristem activity by interactions between the cycline-dependent kinase inhibitor roscovitine and cytokinins

The inhibitors of cytokinin N-glucosylation are known to influence the growth of some plant objects including cotyledons. The use of the plate meristem of zucchini cotyledon as an experimental system allowed us to study for the first time the way in which the changes in the cell division are integrated in this growth reaction. Roscovitine, a potent inhibitor of cytokinin N-glucosylation and cycline-dependent kinases, did not show to have an effect on the meristem activity when applied in 100 μM to cultivated zucchini cotyledons, and acted as an inhibitor in concentrations higher than 400 μM. A 200 μM roscovitine stimulated both palisade cell division and growth. In different seed batches, 400 μM roscovitine acted as a stimulator or an inhibitor. A much stronger stimulating effect on growth and cell division was observed after application of benzyladenine (BA, 10 μM). In contrast to BA, roscovitine provoked a formation of principally flat lamina. In combined treatments, it lowered the stimulating effect of BA; 400 μM roscovitine combined with BA severely suppressed the growth and division activity. This cellular behavior and changes in cotyledon growth could be due to the roscovitine-provoked changes in endogenous cytokinin levels via the inhibition of cytokinin N-glucosylation. Roscovitine-caused stimulation of cell growth and division is stronger in the marginal meristem than that registered in central regions of the cotyledon blade. In this region it also changed the pattern of cell division and lowered the adhesion between the clusters, which enhanced the appearance of local ruptures of the cotyledon edges. The first palisade layer of the plate meristem of cultured zucchini cotyledons, the natural mono-layer of proliferating palisade cells, may be used for screening the inhibitors of cycline-dependent kinases and cytokinin N-glucosylation with regard to their effects on cell division and growth.     

Effects of Chromosome Underreplication on Cell Division in Escherichia coli

The key processes of the bacterial cell cycle are controlled and coordinated to match cellular mass growth. We have studied the coordination between replication and cell division by using a temperature-controlled Escherichia coli intR1 strain. In this strain, the initiation time for chromosome replication can be displaced to later (underreplication) or earlier (overreplication) times in the cell cycle. We used underreplication conditions to study the response of cell division to a delayed initiation of replication. The bacteria were grown exponentially at 39°C (normal DNA/mass ratio) and shifted to 38 and 37°C. In the last two cases, new, stable, lower DNA/mass ratios were obtained. The rate of replication elongation was not affected under these conditions. At increasing degrees of underreplication, increasing proportions of the cells became elongated. Cell division took place in the middle in cells of normal size, whereas the longer cells divided at twice that size to produce one daughter cell of normal size and one three times as big. The elongated cells often produced one daughter cell lacking a chromosome; this was always the smallest daughter cells, and it was the size of a normal newborn cell. These results favor a model in which cell division takes place at only distinct cell sizes. Furthermore, the elongated cells had a lower probability of dividing than the cells of normal size, and they often contained more than two nucleoids. This suggests that for cell division to occur, not only must replication and nucleoid partitioning be completed, but also the DNA/mass ratio must be above a certain threshold value. Our data support the ideas that cell division has its own control system and that there is a checkpoint at which cell division may be abolished if previous key cell cycle processes have not run to completion.     

Gas vesicle formation and buoyancy regulation in Pelodictyon phaeoclathratiforme (Green sulfur bacteria)

Gas vesicle formation and buoyancy regulation in Pelodictyon phaeoclathratiforme strain BU1 (Green sulfur bacteria) was investigated under various laboratory conditions. Cells formed gas vesicles exclusively at light intensities below 5 mol · m^-2 · s^-1 in the stationary phase. No effect of incubation temperature or nutrient limitation was observed. Gas space of gas vesicles occupied always less than 1.2% of the total cell volume. A maximum cell turgor pressure of 330 kPa was determined which is comparable to values determined for cyanobacterial species. Since a pressure of at least 485 kPa was required to collapse the weakest gas vesicles in Pelodictyon phaeoclathratiforme, short-term regulation of cell density by the turgor pressure mechanism can be excluded.Instead, regulation of the cell density is accomplished by the cease of gas vacuole production and accumulation of carbohydrate at high light intensity. The carbohydrate content of exponentially growing cells increased with light intensity, reaching a maximum of 35% of dry cell mass above 10 mol · m^-2 · s^-1. Density of the cells increased concomitantly. At maximum density, protein and carbohydrate together accounted for 62% of the total cell ballast. Cells harvested in the stationary phase had a significantly lower carbohydrate content (8–12% of the dry cell mass) and cell density (1010–1014 kg · m^-3 with gas vesicles collapsed) which in this case was independent of light intensity. Due to the presence of gas vesicles in these cultures, the density of cells reached a minimum value of 998.5 kg · m^-3 at 0.5 mol · m^-2 · s^-1.The cell volume during the stationary phase was three times higher than during exponential growth, leading to considerable changes in the buoyancy of Pelodictyon phaeoclathratiforme. Microscopic observations indicate that extracellular slime layers may contribute to these variations of cell volume.