MITOSIS AND CYTOKINESIS IN ASTEROMONAS GRACILIS, A WALL-LESS GREEN MONAD1

Asteromonas gracilis Artari remains motile throughout cell division. Basal bodies separate and replicate at prophase. They are located lateral to the poles of the closed metaphase spindle. Kinetochores appear at late metaphase. Chromosomes move to the poles and extensions of the nuclear envelope develop into the pyrenoid at anaphase. The interzonal spindle disintegrates at telophase and a diffuse phycoplast is present. Cytokinesis proceeds rapidly from the anterior region of the cell. Newly formed daughter cells have four narrow-banded rootlets and both distal and proximal fibers connect the basal bodies. Features of cell division in Asteromonas are compared to those in other algae, particularly Dunaliella and Chlamydomonas.     

A RHIZOPLAST IN CARTERIA RADIOSA (CHLOROPHYCEAE)12

A rhizoplast or rhizoplast-like structure was observed with the electron microscope in Carteria radiosa. The cross-banded structure extends from the proximal end of each of at least 2 of the basal bodies and extends toward, although does not make contact with, the nucleus. The rhizoplast terminates in a ribosome-free area composed of fine granules and microfibrils. This is the first ultrastructural verification of a rhizoplast in a volvocalean alga.     

THE ULTRASTRUCTURE OF MEIOSPORES OF COLEOCHAETE PULVINATA (CHAROPHYCEAE)1

In view of the relative importance of reproductive cell ultrastructure in phylogenetic and systematic studies of green algae, we investigated the fine structure of germinating zygotes and meiospores of Coleochaete pulvinata Braun. Meiospores have a flagellar apparatus very similar to that of zoospores and spermatozoids of the same species. Meiospores differ from zoospores and spermatozoids of C. pulvinata in having pyramidal body scales similar to those present on zoospores of C. scutata. Meiospores of C. pulvinata had as many as twice the number of spline microtubules as zoospores, and four times the number present in splines of spermatozoids of the same species. Developing meiospores of C.pulvinata, like those of other Coleochaete species, are individually surrounded by chamber walls. These differed from vegetative cell walls in lacking plasmodesmata. Moreover, the chamber walls in germinating zygotes of C.pulvinata stained a cobalt blue color with resorcinal blue, and fluoresced yellow in the presence of aniline blue, thus exhibiting the staining characteristics of callose. In location, morphology and presence of callose, chamberwalls resemble “special walls” of land plants, they may represent a charophycean spore development preadaptation useful in the evolution of walled spores characteristic of land     

Lipid droplets in the adrenal cortex of the rat. Preservation after tannic acid—paraformaldehyde—glutaraldehyde fixation and extraction during staining

Adrenocortical tissue from the rat was fixed in glutaraldehyde-paraformaldehyde-tannic acid with or without potassium pyroantimonate. An electron opacity was observed in lipid droplets from unstained sections of tissue with or without antimonate in the fixative and is most likely attributable to inclusion of tannic acid in the fixative. The opacity was largely removed after staining with uranyl acetate in absolute methanol followed by lead citrate. Removal of the opacity is attributable to staining in lead citrate, not uranyl acetate, because highly basic solution without lead also removes the density. An electron-opaque rim is present at the interface of lipid droplet and cytoplasm, although no distinct membranous structure is observable. The rim may correspond to myelin-like structures seen sometimes in lipid droplets from adrenocortical cells fixed by routine procedures employing pre-fixation with glutaraldehyde and post-fixation with osmium tetroxide. Results of this study point to the conclusion that ultrathin sections should be examined unstained in the validation of a new regime for processing tissues in electron microscopy.     

Ultrastructure of the spermatozoid in Halimeda tuna (chlorophyceae)

The mature Halimeda tuna spermatozoid was studied under the electron microscope. It is pear-shaped and biflagellate. The nucleus is an uncondensed structure except for a few opaque (chromatin) aggregations and shows a rounded profile. The endoplasmic reticulum is a rather well-developed system of rough cisternae engaged in synthesis and storage of proteins. Free ribosomes are numerous. A large mitochondrial apparatus shaped like a horseshoe lies in the anterior gamete region. Only one single plastid is found, and it exhibits a deeply indented outline, a partially structured matrix, osmiophilic globules, and three to four starch grains. The axoneme pattern is 9 + 2. B tubules show septate lumina. A peculiar structure provided with a three-layered shell covering materials of varying electrondensity lies on the upper surface of both basal bodies. The flagellar root system exhibits a cruciate pattern and sets having an inconstant number of microtubules – ie, three, four, or five.     

Abscisic Acid Content of Algae under Stress*

The distribution of the phytohormone, abscisic acid (ABA), within the phylum of Phycophyta was investigated by an enzyme-linked immunoassay (ELISA). Of 64 algal species tested (originating from 9 divisions, 20 classes and 36 orders, including procaryotes) all species contained ABA, whereas no ABA could be detected in the bacteria Escherichia coli, Rhodospirillum rubrum, and Halobacterium halobium. It is concluded that ABA is universally distributed within the algal kingdom and is not restricted to cormophytes. The ability to synthesize ABA must have been developed even within the procaryotes. The physiological role of ABA in some selected algae was studied by investigating 1. the distribution of ABA between the cells and the culture medium, 2. the responses of endogenous ABA to stress, 3. the synthesis of ¹⁴C-ABA from externally applied ¹⁴C-mevalonic acid, 4. the metabolism of ABA, 5. the effect of externally applied ABA on various physiological reactions of the algae, and the effect of norflurazon on ABA content. ¹⁴C-mevalonic acid served as precursor of ¹⁴C-ABA synthesis in Dunaliella cells and ABA was metabolised to the same products which have been observed in higher plants. In D. parva the internal ABA level increased upon hyperosmotic salt shocks, and in D. acidophila upon alkalization of the medium. Norflurazon caused an increase of ABA content in Dunaliella. Externally applied ABA did not affect photosynthesis, respiration and K⁺ content of the cells. The permeability of the plasma membrane of D. acidophila to water was slightly decreased by ABA. The possible physiological function of ABA in algae is discussed.     

Occurrence of jasmonic acid in Dunaliella (Dunaliellales, Chlorophyta)

The occurrence of jasmonic acid and related compounds in Dunaliella species was investigated using gas-liquid chromatography/mass spectroscopy (GCY MS). Jasmonic acid was identified in the ethyl acetate soluble-acidic fraction of Dunaliella tertiolecta and Dunaliella salina (Dunal) Teodoresco, The concentration of jasmonic acid in D. salina. which is extremely halophilic, was much higher than that in D. tertiolecta Butcher, These results indicate that jasmonic acid might play an important role in salt-tolerance in Dunaliella.     

Long-term responses of the green-algal lichen Parmelia caperata to natural CO2 enrichment

Acclimation to elevated CO₂ was investigated in Parmelia caperata originating from the vicinity of a natural CO₂ spring, where the average daytime CO₂ concentration was 729 ± 39 μmol mol⁻¹ dry air. Thalli showed no evidence of a down-regulation in photosynthetic capacity following long-term exposure to CO₂ enrichment in the field; carboxylation efficiency, total Ribulose bisphosphate carboxylase/oxygenase (Rubisco) content, apparent quantum yield of CO₂ assimilation, and the light-saturated rate of CO₂ assimilation (measured under ambient and saturating CO₂ concentrations) were similar in thalli from the naturally CO₂ enriched site and an adjacent control site where the average long-term CO₂ concentration was about 355 μmol mol⁻¹. Thalli from both CO₂ environments exhibited low CO₂ compensation points and early saturation of CO₂ uptake kinetics in response to increasing external CO₂ concentrations, suggesting the presence of an active carbon-concentrating mechanism. Consistent with the lack of significant effects on photosynthetic metabolism, no changes were found in the nitrogen content of thalli following prolonged exposure to elevated CO₂. Detailed intrathalline analysis revealed a decreased investment of nitrogen in Rubisco in the pyrenoid of algae located in the elongation zone of thalli originating from elevated CO₂, an effect associated with a reduction in the percentage of the cell volume occupied by lipid bodies and starch grains. Although these differences did not affect the photosynthetic capacity of thalli, there was evidence of enhanced limitations to CO₂ assimilation in lichens originating from the CO₂-enriched site. The light-saturated rate of CO₂ assimilation measured at the average growth CO₂ concentration was found to be significantly lower in thalli originating from a CO₂-enriched atmosphere compared with that of thalli originating and measured at ambient CO₂. At lower photosynthetic photon flux densities, the light compensation point of net CO₂ assimilation was significantly higher in thalli originating from elevated CO₂, and this effect was associated with higher usnic acid content. Received: 8 May 1998 / Accepted: 22 January 1999     

The size and turnover of the glycerol pool in Dunaliella

Abstract The rate of incorporation of ¹⁴C from CO₂ into glycerol, and the amount of glycerol in cells of Dunaliella tertiolecta was determined at constant salinity at two representative concentrations of NaCl. From these data, the rate of synthesis and turnover of the glycerol pool was determined. The half-time for turnover of the glycerol pool was of the order of 1 h in 170 mol m^?3 NaCl and almost 6h for 700 mol m^?3 NaCl. These results indicate that turnover of the glycerol pool in Dunaliella is relatively slow under steady-state conditions. Synthesis and dissimilation of glycerol do not apparently constitute a metabolic cycle in the conventional sense. Rather, glycerol metabolism resembles that of the storage polysaccharides which arc commonly produced and degraded by different pathways.     

Cell death upon H(2)O(2) induction in the unicellular green alga Micrasterias

In the present study, we investigated whether the unicellular green alga Micrasterias denticulata is capable of executing programmed cell death (PCD) upon experimental induction, and which morphological, molecular and physiological hallmarks characterise this. This is particularly interesting as unicellular freshwater green algae growing in shallow bog ponds are exposed to extreme environmental conditions, and the capacity to perform PCD may be an important strategy to guarantee survival of the population. The theoretically 'immortal' alga Micrasterias is an ideal object for such investigations as it has served as a cell biological model system for many years and details on its growth properties, physiology and ultrastructure throughout the cell cycle are well known. Treatments with low concentrations of H(2)O(2) are known to induce PCD in other organisms, resulting in severe ultrastructural changes to organelles, as observed in TEM. These include deformation and part disintegration of mitochondria, abnormal dilatation of cisternal rims of dictyosomes, occurrence of multivesicular bodies, an increase in the number of ER compartments, and slight condensation of chromatin. Additionally, a statistically significant increase in caspase-3-like activity was detected, which was abrogated by a caspase-3 inhibitor. Photosynthetic activity measured by fast chlorophyll fluorescence decreased as a consequence of H(2)O(2) exposure, whereas pigment composition, except for a reduction in carotenoids, was the same as in untreated controls. TUNEL positive staining and ladder-like degradation of DNA, both frequently regarded as a hallmark of PCD in higher plants, could only be detected in dead Micrasterias cells.     

THE FLAGELLAR APPARATUS OF ENTOCLADIA VIRIDIS MOTILE CELLS,AND THE TAXONOMIC POSITION OF THE RESURRECTED FAMILY ULVELLACEAE (ULVALES,CHLOROPHYTA)1

The flagellar apparatuses of the quadriflagellate zoo-spores and biflagellate female gametes of the marine chaetophoracean alga Entocladia viridis Reinke are significantly different from those of algae belonging to Chaetophoraceae sensu stricto, but closely resemble those of ulvacean genera. These differences permit the taxonomic reassignment of certain marine chaetophoracean genera and an evaluation of the flagellar apparatus features used to characterize the class Ulvophyceae. Critical features of the zoospore include arrangement of the four basal bodies into an upper and a lower pair with the proximal ends of the upper basal bodies overlapping, terminal caps, proximal sheaths connected to one another by striated bands, and a cruciate microtubular rootlet system having a 3-2–3-2 alternation pattern and striated microtubule-associated components that accompany the two-membered rootlets. An indistinct distal fiber occurs just anterior to the basal bodies, and is closely associated with the insertion into the flagellar apparatus of the three-membered rootlets. The flagellar apparatus demonstrates 180° rotational symmetry, and its components show counterclockwise absolute orientation when viewed from above. Newly described features include the prominently bilobed structure of the terminal caps on the upper basal body pair, and the presence of both a granular zone and an additional single microtubule anterior to each of the four rootlets, an arrangement termed the “stacked rootlet configuration.” Rhizoplasts were not observed and are presumed to be absent. The gamete is identical, except for the absence of the lower basal body pair and the presence of an electron-dense membrane associated structure that resembles the mating structure found in Ulva gametes. These findings, correlated with life history data, sporangial and gametangial structure and developmental patterns, chloroplast pigment arrays, and vegetative cell ultrastructural features, compel the removal of Entocladia viridis and similar members of the marine Chaetophoraceae to a separate family, the Ulvellaceae. The latter is referred to the order Ulvales of the Ulvophyceae. The counterclockwise absolute orientation of components, and terminal caps, may be the most consistent flagellar apparatus features of ulvophycean green algae, while variations in other features previously considered diagnostic for the Ulvophyceae may serve instead to identify discrete lineages within this class.