Cell size and chloroplast size in relation to chloroplast replication in light-grown wheat leaves

As part of an investigation into the control of chloroplast replication the number and size of chloroplasts in mesophyll cells was examined in relation to the size of the cells. In first leaves of Triticum aestivum L. and T. monococcum L. the number of chloroplasts in fully expanded mesophyll cells is positively correlated with the plan area of the cells. The linear relationship between chloroplast number per cell and cell plan area is also consistent over a fivefold range of cell size in isogenic diploid and tetraploid T. monococcum. In T. aestivum the chloroplast number per unit cell plan area varies among cells in relation to the size of the chloroplasts. Those cells containing chloroplasts with a relatively small face area have a correspondingly higher density of chloroplasts, and consequently, the total chloroplast area per unit cell plan area is very similar in all the cells. The results indicate that the proportion of the cell surface area covered by chloroplasts is precisely regulated, and that this is achieved during cell development by growth and replication of the chloroplasts.     

杉木花粉发育过程中形态结构的变化研究

该研究以扬州地区成年杉木为材料,采用半薄切片法以及扫描电镜和透射电镜观察法,对杉木花粉发育过程进行了观察。结果表明:杉木的造孢细胞形成于10月,持续至翌年的1月底至2月初。造孢细胞位于小孢子囊最里面,呈多边形,数量多且彼此排列紧密;细胞壁很薄,细胞核较大,内含丰富的细胞器和脂类物质。造孢细胞发育成熟后彼此分离形成小孢子母细胞。小孢子母细胞的减数分裂开始于2月下旬,经两次减数分裂,先后形成二分染色体和四分染色体。小孢子从四分体释放后,体积迅速增大,发育形成花粉,这个过程中,伴随着明显的营养物质变化。杉木成熟花粉壁由薄壁区和厚壁区两部分组成,薄壁区中央有一个乳头状突起,突起上有一萌发孔。成熟花粉外壁着生了许多瘤状颗粒。该研究结果为杉木生殖发育、有性繁殖以及系统演化研究提供了重要依据。     

Apomixis in plants: structural and functional aspects of diplospory inPoa nemoralis andP. palustris

Summary Data on structural and functional aspects of mitotic diplospory and later stages of apomictic seed formation are reported forPoa palustris andP. nemoralis. In this study, the plant material of two Russian populations ofP. nemoralis andP. palustris were used for transmission electron microscope observations. Seed formation was investigated by phase contrast microscopy in two populations ofP. nemoralis collected in The Netherlands. The processes of transformation of the megasporocytes to the megaspore mother cells of diplosporous embryo sacs, and thereafter to one- and two-nucleate diplosporous embryo sacs (Antennaria type) were characterized by an increase of cell size, structural and functional reorganization of the nucleus, nucleolus, and cytoplasm, and cell isolation as a result of thickening of the cell wall. These were accompanied by an increase in the cell metabolic activity inferred from visual evidence of the activation of nucleus, nucleolus, endoplasmic reticulum, dictyosomes, mitochondria, and from the appearance of a dense population of ribosomes and polysomes. The diplosporous embryo sac of the Antennaria type was characteristic for bothP. nemoralis andP. palustris. No signs of the presence of synaptonemal complexes were observed during the process of diplosporous-embryo-sac megaspore mother cell differentiation and division. About 90–95% of the diploid egg cells of diplosporous embryo sacs were able to produce apomictic embryos. These embryos developed before anthesis. However, many of them degenerated at the globular stage because of lack of endosperm. The ultrastructural events occurring during the process of diplospory of apomictic species, and meiosis and megagametogenesis of sexually reproduced plants are discussed.Abbreviations DMC megaspore mother cell of diplosporous embryo sac - TEM transmission electron microscopy - ER endoplasmic reticulum     

On the precision and accuracy achieved by Escherichia coli cells at fission about their middle

Length and width of each of the prospective siblings of constricted Escherichia coli cells from different strains and culture conditions were measured from electron micrographs. The data were statistically analyzed to investigate how equally the length and volume of one cell was divided into two. The analysis showed that, for all cultures, bipartition is unbiased or very nearly so, i.e. sibling cells were on the average equally long and large. The precision of bipartition attained by the cells was usually high; it was related to the average cell shape (length/width): slender E. coli cells divided into two less precisely than squat cells. Absolute size, growth rate and strain specificity affected the precision of bipartition only indirectly, i.e. in as much as they influenced cell shape.     

Patterns of asexual reproduction in <Emphasis Type="Italic">Nannochloris bacillaris</Emphasis> and <Emphasis Type="Italic">Marvania geminata</Emphasis> (Chlorophyta,Trebouxiophyceae)

Non-flagellated vegetative green algae of the Trebouxiophyceae propagate mainly by autosporulation. In this manner, the mother cell wall is shed following division of the protoplast in each round of cell division. Binary fission type Nannochloris and budding type Marvania are also included in the Trebouxiophyceae. Phylogenetic trees based on the actin sequences of Trebouxiophyceae members revealed that the binary fission type Nannochloris bacillaris and the budding type Marvania geminata are closely related in a distal monophyletic group. Our results suggest that autosporulation is the ancestral mode of cell division in Trebouxiophyceae. To elucidate how non-autosporulative mechanisms such as binary fission and budding evolved, we focused on the cleavage of the mother cell wall. Cell wall development was analyzed using a cell wall-specific fluorescent dye, Fluostain I. Exfoliation of the mother cell wall was not observed in either N. bacillaris or M. geminata. We then compared the two algae by transmission electron microscopy with rapid freeze fixation and freeze substitution; in both algae, the mother cell wall was cleaved at the site of cell division, but remained adhered to the daughter cell wall. In N. bacillaris, the cleaved mother cell wall gradually degenerated and was not observed in the next cell cycle. In contrast, M. geminata daughter cells entered the growth phase of the next cell cycle bearing the mother and grandmother cell walls, causing the uncovered portion of the plane of division to bulge outward. Such a delay in the degeneration and shedding of the mother cell wall probably led to the development of binary fission and budding.     

Improved electrotransformation frequencies of Corynebacterium glutamicum using cell-surface mutants

The electrotransformation efficiency for homologously- and heterologously-derived plasmid DNA was determined for two families of Corynebacterium glutamicum strains derived from ATCC13059 (AS019 and auxotrophic, cell surface mutants MLB133 and MLB194) and ATCC13032 (parent strain and restriction-minus mutants RM3 and RM4), following their growth in LBG supplemented with glycine plus isonicotinic acid hydrazide (INH). Electrotransformation efficiencies of MLB133 were up to 100-fold higher than for strain ASO19 and, when using heterologously-derived plasmid DNA, MLB133 showed efficiencies comparable to or better than strains RM3 and RM4, demonstrating the relative importance of cell surface structures in impeding DNA uptake in C. glutamicum. Transmission electron microscopy analysis of cell surface structures showed that strain MLB133 has a thin cell wall relative to AS019 and growth in either glycine or INH further diminished its thickness. Both RM3 and RM4 were more sensitive to INH than ATCC13032 and growth in glycine plus INH further improved transformation efficiency. The mycolic acid composition of these strains is described and the impact of glycine and INH on this is reported.     

Purkinje cell size is reduced in cerebellum of patients with autism

1. The authors' goal was to compare the size and density of Purkinje cells in the cerebellum of subjects with and without autism. Blocks of cerebellum were dissected at autopsy from the brains of age, sex- and postmortem-intervaled (PMI) groups of autistic and normal control individuals (N = 5 per group). Frozen, unfixed blocks were sectioned and stained with 1% cresyl violet.2. The linear, molecular, granular densities and cross-sectional area of Purkinje cells were measured using computer-assisted image analysis. The average cross-sectional areas of Purkinje cells of the patients with autism were smaller by 24% when compared to the normal subjects. Two of the five autistic subjects had mean Purkinje cell sizes that corresponded to greater than 50% reduction in size. There was a substantial effect size difference in Purkinje cell size (² = 0.29) between control and autistic brains (F(1, 8) = 3.32, P = 0.106). No differences in Purkinje cell densities were observed between the two groups.3. These data indicate the possibility of Purkinje cell atrophy in autism with significant neurohistological heterogeneity among individuals diagnosed with this disorder.     

Changes of cell size distribution during the batch culture of Arthrobacter strain PI/1-95

The size distributions of an arthrobacter, approximately 1 m in diameter, were analysed using a Coulter® Multisizer II instrument thereby making it possible to distinguish between the different stages in the morphological cycle. The results indicated that at the beginning of exponential phase a shift occurred from large to smaller cells, the cell size distributions in both categories were asymmetric, skewed towards higher values than the means. During the course of the batch culture the cells in the larger class decreased in size and the two classes coalesced. The size distribution of the smaller class exhibited an additional shift towards smaller cells. After which neither the number of classes nor the size distribution changed. It was concluded that the Coulter® Multisizer II instrument provides a feasible method to extract information not only about mean cell size but also about cell size distributions.     

Cellular aggregation in Chlamydomonas (Chlorophyceae) is chimaeric and depends on traits like cell size and motility

How the variation in phenotypic traits like cell size and motility impacts predator-induced cellular aggregation is not known. Furthermore the genetic composition of cell groups in mixed populations of Chlamydomonas has not been investigated. An examination of these two questions will not only enhance our understanding of Chlamydomonas ecology, but also shed light on the primordial steps before integrated multicellular groups were established. Group living comes with viability and reproductive costs and it is not known how these are shared if groups are genetically heterogeneous. We observed that the natural predator Peranema trichophorum (Euglenoidea) induced clumping in Chlamydomonas. When co-cultured with P. trichophorum cells protected themselves by forming facultative groups (reverting back to a unicellular lifestyle once predators were removed). The dynamics of group formation in different Chlamydomonas species and strains correlated with cell size and swimming speed. Small or less motile strains aggregated more readily than large, fast-swimming ones. Interestingly, Chlamydomonas groups were both intra-species and inter-species chimaeric. This suggests that the predator-induced group formation in Chlamydomonas involved cells coming together rather than staying together and during aggregation cells showed little or no discrimination between self and non-self. These data demonstrate that the dynamics of cell aggregation, in unicellular volvocines at least, depends on phenotypic traits like cell size and motility and high genetic relatedness is not mandatory at this initial stage. These findings further our understanding of aggregation in mixed Chlamydomonas populations and have implications for understanding the very first steps on the road to simple multicellularity.     

Parasitism of aphid embryos by Aphidius smithi: some effects of extremely small host size

Embryos of the pea aphid, Acyrthosiphon pisum (Harris), were successfully parasitized by Aphidius smithi Sharma & Subba Rao while still inside their mother. Parasites from embryos were much smaller, laid fewer eggs, but survived as long as their counterparts that had developed in adult viviparous aphids. Lifetime fecundity was positively correlated with wasp size, as measured by dry weight at the time of death. A. smithi females produced the same offspring sex ratio at eclosion on both host classes, although emergence was significantly lower from mummified adult aphids than from embryos. The data are discussed with regard to progeny and sex allocation decisions by solitary hymenopterous parasites. It is suggested that, in A. smithi, the decision to lay a fertilized or unfertilized egg is influenced by the host's overall size, rather than by the size and quality of a smaller target area inside the host, such as embryos.     

Control of the timing of cell division in fission yeast : Cell size mutants reveal a second control pathway

Strains of Schizosaccharomyces pombe carrying the wee 1 mutation divide at a reduced cell size compared with the wild-type. In this paper, we investigate the mechanism which determines the time of division and cell size at division in wee 1 strains, using three experimental approaches. The evidence suggests that the wild-type control (a cell size control over entry into nuclear division) is absent in wee 1 strains. Instead, a mechanism operates which comprises a cell size control over the initiation of S phase plus a minimum incompressible period in G2 (“timer”) from S phase to nuclear division. The elements of this second control mechanism exist in wild-type cells, though the control is not normally expressed. In particular, the G2 interval in wild-type cells is normally longer than that in wee 1 cells, but can be reduced to this minimum value by delaying S phase. Thus there are two independent controls over entry into nuclear division, one of which operates in wild-type, and the other in wee 1 cells.     

鱼腥草花粉母细胞减数分裂特征及其育性研究

为深入了解鱼腥草花粉母细胞的减数分裂特征与花粉育性的关系,该研究采用卡宝品红染色法对2个鱼腥草居群花粉母细胞的减数分裂过程进行观察,并采用氯化三苯基四氮唑(TTC)染色法、I2-KI染色法、B-K培养基培养法及荧光显微镜观察法来检测鱼腥草花粉的活力及萌发率。结果发现:(1)鱼腥草减数分裂的进程与花序大小、花药颜色、花药长度均有密切的关系。(2)2个居群的鱼腥草中花粉母细胞减数分裂过程正常占88.2%,有11.8%的花粉母细胞减数分裂异常。(3)减数分裂异常表现在减数分裂过程中出现微核、落后染色体、染色体桥、不均等分离、多分体等现象,并发现在二分体阶段及单核花粉发育过程中存在细胞融合。(4)2个居群的鱼腥草花粉活力均不超过1.5%,花粉几乎不萌发。研究认为,鱼腥草花粉育性低的主要原因是单核花粉的发育过程异常,而非鱼腥草花粉母细胞减数分裂异常所致。     

Cell morphology and growth characteristics ofPorphyridium aerugineum (Rhodophyta)

Four unialgal strains of the freshwater coccoid red algaPorphyridium aerugineum Geitler were cultivated under laboratory conditions. Cell morphology was studied with the light microscope. The cell surface was examined by means of electron microscopy in order to contribute to the knowledge of polysaccharide sheaths and cytoplasmic membranes. Optimum growth conditions were determined. The range of cell sizes and the average dry masses of single cells were compared in all four strains cultivated at exactly defined temperatures and irradiances. Photosynthetic pigment maxima were measured in intact cells. The red-coloured phycobiliprotein phycoerythrin was not found in any of the examined strains.Dedicated to Prof. DrLothar Geitler on the occasion of the 90th anniversary of his birthday.     

Effects of nitrate treatment on a mixed species, oil field microbial biofilm

Biofilms of bacteria, indigenous to oil field produced water, were grown in square section, glass capillary flow cells at 45 °C. Initially, in situ image analysis microscopy revealed predominantly coccoid bacteria (length-to-width ratio measurements (l _c:w _c) of bacterial cells gave a mean value of 1.1), while chemical measurements confirmed sulphate reduction and sulphide production. After nitrate ion addition at 100 and 80 mg/l, in the two repeat experiments respectively, the dominance of rod-shaped bacteria (mean l _c:w _c = 2.8) was observed. This coincided with the occurrence of nitrate reduction in the treated flow cells. Beneficially, no significant increase in biofilm cover was observed after the addition of nitrate. The dominant culturable nitrate-reducing bacterium was Marinobacter aquaeolei. The l _c:w _c ratio measured here concurs with previously reported cell dimensions for this organism. Several Marinobacter strains were also isolated from different oil fields in the North Sea where nitrate treatment has been applied to successfully treat reservoir souring, implying that this genus may play an important role in nitrate treatment.     

The genesis of intercellular spaces in developing leaves ofPhaseolus vulgaris L.

Summary Observations by light, transmission electron and scanning electron microscopy have shown that intercellular spaces (ICS) are formed schizogenously in expanding leaves ofPhaseolus vulgaris. ICS formation occurs in predictable positions at the junctions between three or more cells, and follows three phases of development. The first, initiation, phase occurs soon after cell division, and is marked by the formation of an electron-dense osmiophilic body, probably proteinaceous, at the end of the cell plate/middle lamella of the daughter cell wall and across the adjacent piece of the primary wall of the mother cell. This part of the mother cell wall is digested, involving cellulolysis. The second phase, of cell separation, is marked by the first appearance of the ICS. InPhaseolus primary leaves this phase begins about day 3 after sowing, at which time the leaf area is about 1 cm². In the final enlargement phase, lysis of cell wall material continues in the region of the middle lamella, and mechanical tensions arising from the rapid expansion of the lamina lead to further separation of the mesophyll cells so that spaces enlarge and merge.     

Characterization of a major cell wall antigen and potential adhesin in three strains of Candida albicans

Selected strains of Candida albicans were examined to reveal the surface antigenicity and biochemical nature of major cell wall proteins that also were shown to serve as cellular adhesins on human buccal epithelial cells. Confirmation of the adhesive properties of these cells was made by scanning electron microscopy and immunofluorescence microscopy. Particular attention was directed at the clinical isolate KM-302. By means of indirect immunofluorescence staining, the KM-302 blastoconidia absorbed rabbit anti-C. albicans ATCC-32354 serum, revealing specific localization of surface antigens on germ tubes and pseudohyphae. Extracellular polymeric material and the cell wall extract of C. albicans KM-302 blastoconidia were found to contain a major surface antigen of 49 kDa that exhibited 42% adhesion inhibition in vitro. Of considerable significance is that immunogold localization by electron microscopy showed the antigen to be almost exclusively cell wall bound. This major antigen, identified in affinity and gel filtration chromatography fractions, was composed of 4% carbohydrate and 95.7% protein and had an isoelectric point of 6.1. The major antigen also showed a high level of similarity with that of C. albicans strain SC-5314 inasmuch as the major antigen of that strain had carbohydrate and protein compositions of 4 and 95.5%, respectively. Both of these strains also possessed the same percent of adhesion inhibition of human buccal epithelial cells.Abbreviations BECs buccal epithelial cells - CWE cell wall extract - EPP extracellular polymers and proteins - FITC fluorescein isothiocyanate - mAg major antigen - OD ₆₀₀ optical density at 600 nm - PBS phosphate buffered saline - TEM transmission electron microscopy - YNB yeast nitrogen base     

Developmental studies in trebouxioid algae and taxonomical consequences

In the genusTrebouxia (incl.Pseudotrebouxia) two sorts of nonmotile reproductive cells exist: autospores and aplanospores. In subg.Trebouxia small mother cells give rise to 4, 8, or 16 autospores, while comparatively large mother cells develop into zoosporangia or, if the release of zoospores is arrested, into aplanosporangia. Both zoo- and aplanosporangia contain (32) 64 or 128 daughter cells. The transformation of trophic cells into zoo-/ aplanosporangia starts with the formation of a local thickening of the cell wall that marks the prospective opening, and (in most species) with the disappearance of the pyrenoids; sooner or later strong starch deposition can be observed. In subg.Eleutherococcus autospores do not occur; zoo- and aplanosporangia are formed essentially in the same way as in subg.Trebouxia. Differences occur between the form and position of chloroplasts during successive divisions: flattened and parietal in subg.Eleutherococcus, not flattened and ± central in subg.Trebouxia. InEleutherococcus, besides large cells also relatively small cells may produce zoo- or aplanospores.—Dictyosomes could be observed in the living state in representatives of subg.Eleutherococcus under optimal conditions. In trophic cells they are arranged in a group surrounding a hyaline area at the side of the nucleus. In young uninuclear sporangia they are positioned between the nucleus and the local thickening of the cell wall. In somewhat older sporangia they occupy mainly those parts of the nuclear surface which is turned towards the cytoplasmatic cleavage furrow. In subg.Trebouxia dictyosomes could not be observed by light-microscopy. In several species the chloroplast lobation (observed under optimal conditions) differs from that described in the literature.Dedicated to Prof. DrLothar Geitler on the occasion of his 90th birthday, together with cordial thanks for initial scientific guidance and fifty years of stimulation and encouragement.     

Über die Abgrenzung derChlorosarcinales von denChlorococcales

It is proposed to use amongst other characters the type of cell division in order to delimit theChlorosarcinales from theChlorococcales. A definition of the two processes of division occuring in these orders is given. It differs from that of other authors. In theChlorosarcinales only those genera should be assembled in which vegetative daughter cells arise by bipartition followed by firm association of the wall between the daughter cells with that of the mother cell. In contrast, autospores, the vegetative daughter cells of a number ofChlorococcales, develop by multiple division, their cell walls are formed all around the protoplasts and are free from that of the mother cell. The chlorococcalean generaTrebouxia andDictyochloropsis incorporate species which multiply by zoo-, aplano- and autospores as well as others having no autospores. Autospores possibly have arisen more than once during evolution.

     

Effects of temperature on the size and shape of Escherichia coli cells

Two substrains of Escherichia coli B/r were grown to steady-state in batch cultures at temperatures between 22 and 42° C in different growth media. The size and shape of the cells were measured from light and electron micrographs and with the Coulter channelizer. The results indicate that cells are shorter and somewhat thicker at the lower temperatures, especially in rich growth media; cell volume is then slightly smaller. A lower temperature was further found to increase the relative duration of the constriction period. The shapes of the cell size distributions are indistinguishable, indicating that the pattern of growth of the cells is the same at all temperatures. The adaptation of the cells to a temperature shift lasted several generations, indicating that the morphological effects of temperature are mediated by the cell's physiology.     

Cells of Pseudomonas putida and Enterobacter sp. adapt to toxic organic compounds by increasing their size

The phenol-degrading solvent-tolerant bacterium Pseudomonas putida P8 changed its cell shape when grown in the presence of aromatic compounds such as phenol and 4-chlorophenol. The sizes of cells that had been growing after addition of different concentrations of the toxic compounds were measured using a coulter counter that calculates the sizes of the rod-shaped bacteria to diameters of virtual spheres. The cells showed an increase in the diameter depending on the toxic effects of the applied concentrations of both solvents. The same effect was measured for an alkanol degrading bacterium, Enterobacter sp. VKGH12, in the presence of n-butanol. The reaction of the cells to different concentrations of n-butanol was examined by scanning electron microscopy. With this technique it could be shown that the size of the bacteria increased with increasing concentrations of n-butanol. These changes in cell size were dependent on the cellular activity and occurred only after addition of non-lethal concentrations. In the presence of lethal concentrations that completely inhibited cell growth, the cell sizes were similar to those of cells without intoxication. Taking into account the mathematical formula for spherical and cylindrical diameter and surface, respectively, the cells reacted to the presence of organic solvents by decreasing the ratio between surface and volume of the cells and therefore reducing their relative surfaces. As the cell surface and especially the cytoplasmic membrane are the major targets for the toxic effects of membrane-active compounds, this reduction of the relative surface represents an adaptive response to the presence of such compounds.