Studies on the control of cell division in a green alga,Ankistrodemus gracilids (Chlorococcales)

The proliferation patterns of the green alga,Ankistrodesmus gracilis were investigated by electron microscopy. The multichotomically dividing phase, namely the type I phase was characterized by multichotomical nuclear division, whereas the dichotomically dividing phase, i.e., the type II phase was related to dichotomical nuclear division. Type I phase was controlled by a lower cell density and a diurnal light-dark rhythm of LD 14:10. Type II phase was induced by a cell density of more than 1.0×10⁷ cells/ml. Independent of the 2 types of proliferation patterns, the diurnal light-dark rhythms related to the autospore release from a mother cell. The diurnal light-dark rhythm of LD 14∶10 induced the scattered release of autospores. On the other hand, a rhythm of LD 18∶6 induced the release of a couple of tightly adhered autospores from a mother cell which had been observed apparently as a type II cell by light microscopy.     

Studies on the control of cell division in a green alga, Ankistrodesmus gracilis (Chlorococcales) III. Induction synchrony with controlled division patterns

When stagnant cells of Ankistrodesmus gracilis obtained froma standard culture were inoculated into the basal medium atcell densities lower than 1.0 ? 10⁷ cells/ml, cell proliferationoccurred stepwise at time intervals of about 30 hr. At a densityof 5.5 ? 10⁴cells/ml, the increase in cell number per step wasabout 2.7-fold. When inoculated into a glutamine medium thetime interval was 24 hr, and the average increase of cell numberwas about 4-fold. When cells were preincubated at about 5.0? 10⁵ cells per ml in the basal medium for 30 hr, then transferredinto a glutamine or arginine medium at about 7.0 ? 10⁶ cells/ml,synchronous division occurred about 18 hr later with binaryfission or about 33 hr later with multiple fission, respectively. (Received May 16, 1979; )     

DNA Levels in Dividing and Developing Plastids in Expanding Primary Leaves of Avena sativa

The amounts of plastid DNA in the primary leaves of 4-d-oldlight- and dark-grown seedlings of Avena sativa were measuredby microspectrofluorometry using the DNA-fluorochrome DAPI (4',6-diamidino-2-phenylindole). In the light-grown primary leaves (40–45 mm long) therewas a marked increase in DNA level per plastid from 10.2 to18.5 ? 10^–15 g between 2.0 mm and 10 mm from the leafbase, resulting from the rate of plastid DNA synthesis beinghigher than the rate of plastid division. Beyond 30 mm the plastidDNA level was reduced to 14 ? 10^–15g due to chloroplastdivision rates being higher than the rate of plastid DNA synthesis,while from 20 mm plastid DNA levels were constant at 2.2 ? 10^–12g per cell, which corresponds to 16000 plastome copies per cell. Observations of dark-grown leaves establish that, in Avena,light is not necessary for plastid division and the dark-grownleaf cells accumulate higher amounts of plastid DNA than light-grownleaf cells. Plastid nucleoids showed a change of distribution after completionof plastid DNA synthesis in light-grown leaves. A change inthe distribution of plastid nucleoids was also observed duringthe greening of etioplasts of dark-grown leaves while plastidDNA level remained constant. Such changes in plastid nucleoiddistribution appear to be independent of plastid DNA synthesisand correlate with the formation of grana stacks. Key words: Avena sativa, microspectrofluorometry, plastid DNA     

A Cryomicroscopic Study of Cylindrocystis brebissonii De Bary and Two Species of Micrasterias Ralfs (Conjugatophyceae, Chiorophyta) during Freezing and Thawing

The changes in morphology of the unicellular algae Cylindrocystisbrebissonii and two species of Micrasterias during freezingand thawing were observed on a light microscope fitted witha temperature controlled stage. At slow rates of cooling extensiveshrinkage of the protoplast was observed. The response of thecell wall varied with cell-type. In C. brebissonii plasmolysiswas not observed and the cell wall and protoplast shrank together.In Micrasterias the cell wall did not contract and a distinctplasmolysis was observed. Following freezing to and thawingfrom –25?C cells of C. brebissonii were non-viable butremained osmotically responsive. Cooling at faster rates inducedintracellular ice formation in all cell-types. The criticalrate of cooling varied with cell-type and was determined bycell volume and suface area. Intracellular gas bubbles wereobserved during thawing following both rapid and slow cooling. Following cooling in dimethylsulphoxide cells of C. brebissoniiwere protected against freezing injury. The recovery on thawingfrom –196?C being determined by the rate of cooling, anoptimum rate of 1?C min^–1 was observed. During slow ratesof cooling (<2?C min^–1) cells remained unshrunken,at faster rates (10?C min^–1) the loss of cell viabilitywas related to osmotic shrinkage during cooling rather thanto nucleation of intracellular ice. Intracellular ice formationwas observed only following significantly faster rates of cooling(>20?C min^–1). Key words: Cylindrocystis, Micrasterias, cryomicroscopy, freezing injury     

Light dependency of the mating process in Closterium acerosum

Sexual cell division and activation of gametangial cells forconjugation in Closterium acerosum were induced by light. L₂₀₀cells conjugated at maximum level under the following conditions;(i) a light intensity higher than 1,000 lux in a 16-hr lightand 8-hr dark regime and (ii) an illumination time longer than12 hr at 3,000 lux. L₂₀₀ cells also conjugated under continuousillumination at 3,000 lux. The action spectrum for the activation of gametangial cellshad peaks around 450, 611 and 665 nm. 3-(4'-Chlorophenyl)-l,l-dimethylurea (CMU) inhibited the accumulationof carbohydrates and sexual cell division at 10^–5 M andthe activation of gametangial cells for conjugation at 10^–4M. (Received August 15, 1977; )     

Initial Anatomical Changes Associated with Tuber Formation on Single-node Potato (Solanum tuberosum L.) Cuttings: A Re-evaluation

Cell division and cell expansion during early stages of tuberdevelopment were studied using developing axillary buds on single-leafcuttings from potato (Solanum tuberosum L.). Cuttings takenfrom plants induced to form tubers, by short day (SD) treatment,were compared with cuttings from non-induced (long day, LD)plants. In the apical zone of the buds, cell division occurredfrom the first day after cutting, in both LD and SD cuttings.The planes of these divisions were transverse, associated withelongation of the buds. At day 5, a new orientation of celldivision was observed in the subapical zone of SD cuttings only.These divisions were longitudinal, associated with radial growth.Cell expansion occurred in both SD and LD cuttings, and wasnot uniquely related to the onset of tuber formation. Copyright1999 Annals of Botany Company Solanum tuberosum L., potato, tuber formation, cell division, cell expansion.     

Inhibitory effect of fusicoccin on cell division

Rapid interactions in cell division and cytodifferentiationare induced by hormone treatments in dark-cultured explantsof Jerusalem artichoke. Fusicoccin, at concentrations between10^–6 and 10^–5 M, markedly inhibited the division-promotingactivity induced by plant hormones. Further, fusicoccin-treatedmeristematic root tips of Vicia faba and Allium cepa showeda rapid decrease in the mitotic index. Fusicoccin seems to inhibitsome hormone-sensitive processes required during the inductionand regulation of cell division. (Received March 28, 1979; )     

Induction of Cell Division Synchrony and Variation of Cytokinin Contents through the Cell Cycle in Tobacco Cultured Cells

Cell division synchrony was induced in tobacco {Nicotiana tabacum)cultured cells by several treatments. Very high synchrony throughouttwo cell cycles was induced by aphidicolin treatment (inhibitorof DNA polymerase , 10 µg/ml) and by treatment with lowtemperature (4°C) and hydroxyurea (50 µg/ml). Themitotic index reached its maximum (52% and 40% in aphidicolinand hydroxyurea treatments, respectively) at 11 h after removalof the added chemical. During the treatments, the cells werearrested in the G₁/S phase of the cell cycle. In the aphidicolin-inducedsystem, incorporation of ¹⁴C-thymidine confirmed that DNA synthesiswas started immediately after removal of the chemical. The aphidicolin-induced synchronous cells were used to studythe contents of butanol-soluble cytokinins during the cell cycle.Cytokinin contents increased conspicuously at the G₂/M boundary. ¹Present address: Department of Biology, Otsuma Women's University,Chiyodaku, Tokyo 102, Japan. (Received May 14, 1985; Accepted November 8, 1985)     

Cultivation of Rice Protoplasts and Their Transformation Mediated by Agrobacterium Spheroplasts

Improvement of the cultivation of rice (Oryza saliva L.) protoplastsisolated from suspension cultures led to their division at afrequency of 5 to 10%. Rapidly growing colonies were obtainedon a hormone-free medium when Agrobacterium tumefaciens spheroplastswere introduced into the protoplasts by polyethylene glycoltreatment. Opines corresponding to the strains of A. tumefaciensused for the spheroplast treatments were detected in some ofthese colonies at a frequency of about 10^–4. Using radioactiveprecursors, [¹⁴C]--ketoglutaric acid and [³H]-arginine, activitiesof nopaline synthase, a marker enzyme of nopaline-type crowngall, were also detected in some of these clones. These resultsshow that the rice cells were transformed by Ti plasmid introducedby the spheroplast method. (Received September 6, 1985; Accepted January 24, 1986)     

Effect of Laurie Acid on Cell Division, Macromolecular Synthesis and Membrane Lipid Organization in Escherichia coli

Laurie acid (1 mg/ml) sharply suppressed the cell division ofan acrA mutant strain of Escherichia coli K12. However, thewild type acrA^$ strain was resistant to the fatty acid. Capricacid and myristic acid were not so toxic. Laurie acid inhibitedboth DNA and protein synthesis of the acrA mutant strain, withthe former being more sensitive than the latter. On the otherhand, DNA polymerase activity of toluene-treated cells was stimulatedrather than inhibited by the presence of 1 mg/ml of lauric acid.Fatty acid composition of phospholipids in the inner membranewas largely altered by the addition of lauric acid. These resultssuggest that addition of lauric acid to the medium causes adisorganization of the membrane lipids in the acrA mutant celland activities of DNA polymerase and other intramembranous enzymesare consequently inhibited. ¹Present address: Osaka City Institute of Public Health andEnvironmental Sciences. Osaka 543, Japan. (Received January 28, 1983; Accepted November 15, 1983)     

Colony growth and evidence for colony multiplication in Phaeocystis pouchetii (Prymnesiophyceae) isolated from mesocosm blooms

Using well plates of Phaeocystis pouchetii colonies isolatedfrom experimental mesocosms in western Norway, increases incolony size and division were documented. Median longest lineardimensions increased 0–7 µm h^–1; literaturePhaeocystis globosa values are 0.9–4.7 µm h^–1.Ten to twelve percent of colonies divided at rates of 0.21–0.28divisions day^–1. Daughter colonies were 100 µm smallerthan mother colonies. Colonies delayed 3.5–4.9 days tofirst division, compared with literature values of 4–5days for P. globosa. This study provides the first experimentalevidence for colony division of wild P. pouchetii.     

Determination of Volume of Dunaliella Cells by Lithium Dilution Measurements and Derivation of Internal Solute Concentrations

A method has been developed to measure the cell volume of theunicellular green alga Dunaliella parva 19/9 using Li⁺ measurementsonly. Concentrations of internal solutes can also be calculatedif they are assayed in the same samples as Li⁺. We found thatD. parva cells grown in 0.4 kmol m^–3 NaCl have an averageaqueous cell volume of 65.1 ?2.9 µm³, a K⁺ concentrationof 126?6 mol m^–3, a Na⁺ concentration of 11?11 mol m^–3and a glycerol concentration of 615?27 mol m–3 (n= 12).Algae grown in 1.5 kmol m^–3 NaCl have an average aqueouscell volume of 131 ?7.5 µm³, a K⁺ concentration of 109?4mol m^–3, a Na⁺ concentration of 10?39 mol m^–3 anda glycerol concentration of 1 425?59 mol m^–3 (n = 12).These results indicate that D. parva cells adapted to high salinitieshave larger cell volumes than those adapted to lower salinities.However, there is no evidence for a significant difference ininternal Na⁺ concentration, despite the almost 4-fold differencein the concentration of external NaCl. The intracellular glycerolconcentration alone accounts for 65% and 54%, respectively,of the osmotic balance in low and high salt grown cells. Key words: Dunaliella, cell volume, intracellular solutes     

Inhibitory effect of lycorine on cell division and cell elongation

Lycorine, an alkaloid isolated from bulbs of Amarillidaceae,was found to be a powerful inhibitor of cell division and elongation.Adding different concentrations of lycorine from 10^–6M to 10^–4 M in an appropriate growth-medium strongly inhibitedcell division in explants of lettuce pith parenchyma. The sameresult was obtained with liquid yeast cultures growing exponentially. Lycorine-treated meristematic cells of the primary roots ofVicia faba also showed rapid inhibition of the mitotic indexwhile interphase cells increased proportionately. Lycorine alsoinhibited endogenous and auxin-induced cell elongation in Avenacoleoptiles and pea segments. Since both cell division and cell elongation require proteinsynthesis and RNA synthesis, the assumption is that lycorineprobably inhibits one of the two syntheses. ¹This study was supported by a contract between the NationalResearch Council of Italy and University of Bari, Instituteof Botany. (Received November 27, 1972; )     

Pyrimidine Nucleotide Biosynthesis in Vinca rosea Cells: Changes in the Activity of the de novo and Salvage Pathways during Growth in a Suspension Culture

Marked changes in the activity of the ‘de novo’and ‘salvage’ pathways of pyrimidine biosynthesisduring growth of Vinca rosea cells in a batch suspension culturewere observed. The activity of these pathways was investigated by determiningthe contribution of ¹⁴C of [2-¹⁴Cluracil, 12-¹⁴Cluridine. and[6-¹⁴Clorotate to the cell constituents and by measuring theactivity of the several enzymes of these pathways. During the lag phase of the culture, ‘uracil-’ and‘uridine-salvage’ pathways made the predominantcontribution to nucleotide biosynthesis, but, following theinitiation of cell division, the ‘de novo’ pathwayfor nucleotide biosynthesis operated appreciably. These results suggest that nucleotide synthesis during cellgrowth in a suspension culture can be divided into two stages:a ‘turnover stage’, during the lag phase of cellgrowth, and a ‘true biosynthetic stage’, which isinitiated in the cell division phase.     

SYNCHRONIZATION OF BACTERIAL CULTURE BY PREINCUBATION OF CELLS AT HIGH CELL DENSITY

The growth of Escherichia coli strain B in a liquid medium wasfound to cease at a cell density of 5x10⁹ cells per ml. (Thiscritical concentration is designated as the maximum or M-concentration.)Even cells harvested from the logarithmic growth phase couldnot divide at this or higher cell densities. Investigationson the metabolic activities of such cultures, however, showedthat the synthesis of cellular protein and nucleic acid wastaking place under such circumstances, showing that only someprocess (or processes) particularly related to cell divisionwas suppressed at the critical cell concentration in question. This finding led us to devise a new method of synchronizationof E. coli: cells harvested from a logarithmic phase were preincubatedat the critical concentration of 5x10⁹ cells per ml for 45 minutes,and then diluted 100 times with fresh medium. This led to synchronizationof cell division, as shown by a stepwise multiplication in cellnumber. (Received June 20, 1961; )     

Induction of Cell Division in Leaf Cells of Coconut Palm by Alteration of pH and its Correlation with Glyoxalase-I Activity

Cell division in suspension cultures obtained from leaf cellsof coconut was influenced by pH of the culture media. A 3-foldincrease in cell number was obtained at pH 7.0 compared to suspensionsgrowing at pH 5.0. The pH of both cells and media changed after48 h of growth. Internal cell pH showed a significant increasewhen cultures were grown at pH 7.0 and 8.0 and increased onlyslightly at pH 5.0 and 6.0. Glyoxalase-I activity of cells insuspension culture was found to be pH-depcndent, showing maximumactivity at pH 7.0. Glutathione, a co-enzyme for the substratemethylglyoxaJ for glyoxalase-I, produced a 2-fold increase incell number at a concentration of 5 x 10^–3 mol dm ^–3.The polyamine, spermidine, promoted cell division maximallyat a concentration of 10^–6 mol dm^–3. Methylglyoxal-bis(guanylhydrazone), an inhibitor of spermidine biosynthesis,strongly inhibited cell division giving maximum inhibition ata concentration of 3 x 10^–6 mol dm ^–3. These resultsindicate a positive correlation between cell division and glyoxalase-Iactivity. Key words: Cocos nucifera, glyoxalase-I, pH, spermidine     

Promotion of Plant Cell Division by an Epidermal Growth Factor

In some specified treatments, an epidermal growth factor (EGF)promoted adventitious root formation in epicotyl cuttings ofVigna angularis. The number of the roots induced in cuttingstreated with 0.1 mg liter^-1 EGF during the first 24 h and with210^-4 M IAA during the second 24 h was 15% greater than thatof the roots in cuttings treated without EGF and with IAA. Analysisof the optimum timing of EGF application was performed by dividingthe first 24 h period into three sequential 8 h periods (0–8h, 8–16 h and 16–24 h). The most effective timeperiods in terms of the root formation were 8–16 h and16–24 h. The 0–8 h period was ineffective with respectto the formation. When carrot suspension cells were culturedfor 15 days at a very low cell density (1,000 cells/3 ml Murashigeand Skoog's medium) with more than 0.1 mg liter^-1 EGF, cellnumbers were 72% higher than those cultured without EGF. Theseresults suggest that EGF promotes cell division of plants. (Received October 5, 1992; Accepted May 24, 1993)     

Anatomical and physiological aspects of developmental processes of adventitious root formation in Azukia cuttings

In Azukia stem cuttings, root primordia always appeared in theinterfascicular regions between the endodermis and the interfascicularcambium. Transverse cell divisions were observed as the first eventsin the process of root formation. They began to occur 10 hrafter cuttings had been made and were restricted to the interfascicularregions about 1 mm above the basal cut end of the cutting. Ineach of interfascicular region, 10 to 20 cells divided. Transversedivisions were followed by longitudinal divisions, which beganto occur 18 hr after cuttings had been made. The early process of root primordium formation is distinguishedby the following three phases: the first phase during whichno cell division occurs (0–8 hr), the second phase duringwhich transverse cell divisions occur (8–16 hr) and thethird phase during which longitudinal divisions occur (16–24hr). Cuttings in each phase responded differently to test substances. ¹Supported in part by Grant No. 139011 from the Ministry ofEducation, Japan. ² Present address: Junior College of Toyo University, Hakusan,Bunkyo-ku, Tokyo 112, Japan. (Received October 24, 1977; )     

Involvement of Poly(ADP-Ribose) Synthesis in Transdifferentiation of Isolated Mesophyll Cells of Zinnia elegans into Tracheary Elements

The relationship between poly(ADP-ribose) synthesis and cytodifferentiationwas studied in the well characterized Zinnia system, in whichisolated mesophyll cells of Zinnia elegans transdifferentiateinto tracheary elements (TE) in a suspension culture in thepresence of both auxin and cytokinin. The rate of poly(ADP-ribose)synthesis was measured in nuclei isolated from cells that hadbeen induced to undergo transdifferentiation, and activationof such synthesis was observed before the appearance of TE duringculture. In cultures without auxin or cytokinin, poly-(ADP-ribose)synthesis appeared to proceed much more slowly. Treatment of cells with a potent inhibitor of poly-(ADP-ribose)polymerase, namely, 6(5H)-phenanthridinone (PT), resulted inthe blockage of TE formation and a decrease in the frequencyof cell division. PT was very effective in interfering withtransdifferentiation, in particular, when supplied between the24th hour and the 36th hour of culture. Repair-type DNA synthesis,which has been proposed to participate in transdifferentiation,was suppressed by the treatment with PT. These results suggestthat poIy(ADP-ribose) synthesis and subsequent repair-type DNAsynthesis might play a critical role in the transdifferentiationof Zinnia cells. ³Present address: Botanical Gardens, Faculty of Science, Universityof Tokyo, Hakusan, Bunkyo-ku, Tokyo, 112 Japan. ⁴Present address: Department of Chemical and Biological Sciences,Faculty of Science, Japan Women's University, Mejirodai, Bunkyo-ku,Tokyo, 112 Japan.     

Preservation of the Differentiated State of Mesophyll Cells In Vitro

Under a variety of light conditions, and in a modified NT mediumcontaining sucrose as a carbon source, isolated mesophyll cellsof Calystegia sepium started cell divisions after 72 h. Alternately,under light-dark alterations (12: 12 h; 1.35–1.50 W m^–2of cool white light), and culture in the NT medium lacking anorganic carbon source, and made iso-osmotic by the additionof mannitol, the cells preserved their differentiated stateas regards cell size, cell form, colour and size and distributionof chloroplasts for one week. Later on, they changed their characterand cell division was started. Calystegia sepium, mesophyll cells, differentiated state, in vitro culture