BRIEF INCUBATION OF GAMETANGIA-BEARING CAPS IN ANTIBIOTICS ELIMINATES BRANCHING IN PROGENY OF ACETABULARIA ACETABULUM (CHLOROPHYTA)1

Branching of the stalk of Acetabularia acetabulum L. (Silva) was investigated by inbreeding and by a brief treatment of gametangia with a variety of antibiotics. The position of the branch along the stalk varied, implying that branching was not restricted to any one time in development (base is oldest and apex is youngest). The branching phenotype was not inherited in Mendelian fashion. Although three microscopic structures (“bubbles,”“pustules,” and “scars”) occurred on the stalks of cells that had branched, these structures were not statistically correlated with branching in the population (n=699 cells). However, brief treatment of gametangia with a new antibiotic mixture did eliminate all macro- and microscopic structures associated with branching of the stalk in the subsequent generation. We could not fulfill Koch's postulates or provide clear evidence for the pathogenic nature of cell branching. Our brief antibiotic treatment of gametangaa of Acetabularia acetabulum was rapid, had no adverse effects, and virtually eliminated branching (and any potential pathogens) from laboratory cultures in the subsequent generations. Our method allows biochemical and molecular analyses to proceed uncomplicated by the possible presence of other organisms and provides a clean baseline for the future selection of mutations that may induce heritable branching.     

TIMING AND LIGHT REGULATION OF APICAL MORPHOGENESIS DURING REPRODUCTIVE DEVELOPMENT IN WILD-TYPE POPULATIONS OF ACETABULARIA ACETABULUM (CHLOROPHYCEAE)

In the giant unicellular green alga, Acetabularia acetabulum (L.) Silva, development is altered by light. For example, blue light induces the vegetative apex to produce whorls of hairs that encircle the stalk and, later, blue light may trigger reproductive onset. The two goals of this study were to determine when changes in apical shape occur during formation of the reproductive structure, or "cap," and to determine which of these differentiation events require light. The first visible indication of cap initiation was a rounded swelling of the apex, which we call a knob-shaped apex (time = 0 hours). Subsequent changes in shape were a hyaline, knob-shaped apex, reached by 50% of the population 3 h later, and the formation of a whorl of unilobed chambers at 16 h. These chambers became bilobed at 33 h and trilobed at 34 h. Successive sets of cap hairs grew from protuberances found on the surface of the uppermost lobes of the chambers (superior corona). After knob, the remainder of cap formation was largely independent of light. However, the initiation of each set of cap hairs required light. If a recently initiated cap was amputated, the individual recapitulated development, repeating a portion of vegetative morphogenesis (i.e. it made whorls of sterile hairs) before initiating a new cap. The developmental sequence between amputation and initiation of a new cap required light. A model for light-regulated changes in shape at the apex of Acetabularia acetabulum, which integrates whorl and cap formation and encompasses both vegetative and reproductive development of this organism, is presented.     

PHYSIOLOGICAL FACTORS THAT AID DIFFERENTIATION OF ZYGOTES AND EARLY JUVENILES OF ACETABULARIA ACETABULUM (CHLOROPHYTA)

To improve our understanding of early development and to enable standard genetic analyses in Acetabularia acetabulum (L.) Silva, we examined some physiological factors that impact differentiation of young diploid plants. Eighty percent of zygotes differentiated within 6–7 days when they were inoculated at high densities (≥5,000 zygotes·mL⁻¹), and zygote differentiation was independent of the photon flux density of cool white light. In contrast, 90% of juveniles differentiated within 25 days when they were inoculated at medium densities (<1,000 individuals·mL⁻¹) and required >250 μmol·m⁻²·s⁻¹ cool white light. Both zygotes and juveniles differentiated best at an alkaline pH (7.96), when the temperature was between 18.6° and 23.5° C, and when the external potassium concentration was 1–10 mM. Compared to prior publications, adjustment of these parameters reduced the time to differentiation by 25% (from 8 to 6 days) and more than tripled the percentage of zygotes that developed into healthy juveniles (from 23% to ∼80%).     

18S rDNA AND EVOLUTION IN THE DASYCLADALES (CHLOROPHYTA): MODERN LIVING FOSSILS1

Phylogenetic relationships were inferred from parsimony and distance analyses of nuclear small-subunit ribosomal DNA sequences taken from 14 species representing 8 of the 11 extant genera in the Dasycladales. Of 1733 aligned positions, 412 (23.8%) were variable and 251 (61%) of those were phylogenetically informative within the Dasycladales. Secondary structure was analyzed and taken into account during all phases of data analysis. Robustness of the trees was assessed using bootstrap analysis and g₁ statistics of tree-length decay. Strongly supported branches were robust to all methods of analysis regardless of weighting schemes used. The secondary structure of the 18S within the Dasycladales agrees with that of other green algae with the exception of a shared deletion in stemloop E10-1 (ca. 13 nucleotides long), which provides additional support for the uniqueness of this monophyletic group. A molecular clock was calibrated from the dasyclad fossil record and suggests a radiation of the Acetabulariaceae at 120 ± 30 million years (Ma) ago and the Dasycladaceae 215 ± 40 Ma ago. The split of the two lineages from a shared ancestor is estimated at 265 ± 50 Ma ago. Within the Dasycladaceae, Neomeris and Cymopolia are sister taxa, as are Batophora and Chlorocladus. Bornetella groups with the Neomeris and Cymopolia clade in 78% of the bootstrap replicates. Relationships among the Acetabulariaceae show that Acetabularia and Polyphysa do not form monophyletic groups as presently circumscribed. No evidence indicates that Acicularia is the oldest genus. Halicoryne, Chalmasia, and Dasycladus were not included in the analysis. Molecular data provide afresh background perspective from which to discuss the evolution of one of the most ancient lineages of green plants.     

UNCOUPLING OF VARIOUS MEASURES OF GROWTH IN ULVA ROTUXDATA(CHLOROPHYTA) FOLLOWING A LARGE DECREASE IN IRRADIANCE1

A vegetative clone of the chlorophyte macroalga Ulva rotundata Blid. was maintained in an outdoor continuous flow system and subjected to a large decrease in irradiance. Specific growth rates based on changes in fresh (μ_FW) and dry weight (μ_DW) and surface area (μ_SA) were determined using precut disks over the 24 h following a post-sunset transfer from full sunlight (100% I₀) to 9% I₀ All three measures of growth rate were approximately equivalent for untransferred control plants at either limiting (9%) or saturating (100%)I₀. Transferred disks exhibited μ_FW and μ_SA which were slightly lower than 100%I₀ controls and much higher than 9% I₀ controls; μ_DW was nearly identical for transferred disks and 9% I₀ controls. Cell size was unchanged following transfer, indicating that surface area changes reflected a proportional increase in cell number. Cell division therefore continued at a high rate for one day following transfer of U. rotundata to irradiances which are subsaturating for photosynthesis (indicated by μ_Dw). Starch reserves were largely depleted, and the C/N ratio decreased during this period.     

A STUDY OF CELL WALL AND FLAGELLA FORMATION DURING CELL DIVISION IN THE SCALY GREEN ALGA,SCHERFFELIA DUBIA (CHLOROPHYTA)1

The thecate green flagellate Scherffelia dubia (Perty) Pascher divides within the parental cell wall into two progeny cells. It sheds all four flagella before cell division, and the maturing progeny cells regenerate new walls and flagella. By synchronizing cell division, we observed mitosis, cytokinesis, cell maturation, flagella extension, and cell wall formation via differential interference contrast microscopy of live cells and serial thin‐section EM. Synthesis of thecal and flagellar scales is spatially and temporally strictly separated. Flagellar scales are collected in a pool during late interphase. Before prophase, Golgi stacks divide, flagella are shed, the parental theca separates from the plasma membrane, and flagellar scales are deposited on the plasma membrane near the flagellar bases. At prophase, Golgi bodies start to synthesize thecal scales, continuing into interphase after cytokinesis. During cytokinesis, vesicles containing thecal scales coalesce near the cell posterior, forming a cleavage furrow that is initially oriented slightly diagonal to the longitudinal cell axis but later becomes transverse. After the progeny nuclei have moved into opposite directions, resulting in a “head to tail” orientation of the progeny cells, theca biogenesis is completed and flagellar scale synthesis resumes. Progeny cells emerge through a hole near the posterior end of the parental theca with four flagella of about 8 μm long. The precise timing of flagellar and thecal scale synthesis appears to be an evolutionary adaptation in a scaly green flagellate for the thecal condition, necessary for the evolution of the phycoplast and thus multicellularity in the Chlorophyta.     

DIPLOIDY IN DNA CONTENT IN VEGETATIVE CELLS OF CLOSTERIUM EHRENBERGII (CHLOROPHYTA)1

DNA content of the nucleus in the placoderm desmid, Closterium ehrenbergii Meneghini was measured throughout the life cycle by epifluorescence microspectrophotometry after DNA specific dye [4′,6-diamidino-2-phenylindol (DAPI)] staining. Postulating a mean DNA content of gamete nuclei as 1C, the nucleus of a newly divided vegetative cell was 2C. Most vegetative cells in the stage of exponential growth had a DNA content from 2C to 4C, while most in stationary phase, with the highest frequency of zygote formation, were 2C. They became pre-gametes (2C) upon mixing two heterothallic strains. Four gametes were made by a DNA reduction division of each pre-gamete cell. Therefore, there was a nonmeiotic DNA reduction stage by one half. During germination, the zygote underwent meiosis to produce two gones, each of which contained one surviving nucleus (large nucleus) and one degenerating nucleus (small nucleus). The DNA content of these four nuclei was 1C basically. The DNA of the surviving nucleus duplicated to 2C and further quadruplicated to 4C without cell or nuclear division. These two 4C gones had different cell morphology from ordinary vegetative cells. After the first cell division following meiosis, each gone produced two vegetative cells in which the DNA content became 2C to 4C again.     

SPERM BUNDLE-FEMALE SOMATIC CELL INTERACTION IN THE FERTILIZATION PROCESS OF VOLVOX CARTERI F. WEISMANNIA(CHLOROPHYTA)1

We studied the fertilization reaction in Volvox carter f. weismannia (Powers) Iyengar. Tests for a sperm bundle chemoattractant produced by female spheroids were negative. The flagella of the female spheroid were identified as the site of sperm bundle binding. Treatment of female spheroids with trypsin or protease blocked sperm bundle binding, suggesting surface proteins are involved. Sperm bundle binding is not affected by female spheroid age up to 64 h after inversion of the female spheroid. Soybean trypsin inhibitor prevents fertilization pore formation. This suggests that a trypsin-like enzyme, released by the dissociating sperm bundle, is responsible for fertilization pore formation.     

GROWTH AND BRANCHED HYDROCARBON PRODUCTION IN A STRAIN OF BOTRYOCOCCUS BRAUNII (CHLOROPHYTA)1

A strain Botryococcus braunii Kütz. that produces high levels of branched hydrocarbons (botryococcenes) was grown under different environmental conditions to investigate the relationship between growth and hydrocarbon production. Carbon dioxide concentration had the most significant influence on growth; 0.3% CO₂-enriched cultures demonstrated a minimum mass doubling time of ca. 40 h, compared to ca. 6 days for ambient air cultures grown on the same buffered growth medium. The botryococcene fraction, which consisted of 10 identified compounds (C_nH_2n-10; n = 30–34), usually constituted ca. 25–40% of the culture dry weight under various growth regimes, including nitrogen- and/or phosphate-deficiencies. CO₂ enrichment initially favored the production of the lower botryococcenes (C₃₀–C₃₂), whereas relatively slow-growing ambient air cultures accumulated C₃₃ and C₃₄ compounds. Colony color changed in response to different light intensities. High light increased the carotenoid/chlorophyll ratio, which resulted in orange colonies. Cultures exposed to low light intensity appeared green. This change in coloration was reversible over a period of a few days, and at no time were the linear hydrocarbons characteristic of the other form of the alga detected. Ostensible colony color is not, therefore, a reliable indicator of qualitative hydrocarbon content. Sequential solvent extraction experiments indicated that up to ca. 7% of the botryococcene fraction was intracellular and that the remainder was located within the colonial matrix. The internal (cellular) pool principally consisted of C₃₀–C₃₂ botryococcenes, whereas the external (colonial matrix) pool contained >99% of the C₃₃ and C₃₄ compounds, in addition to large amounts of the lower botryococcenes. These results, taken in conjunction with other data, are compatible with the hypothesis that the C₃₀ botryococcene is the precursor, presumably via methylation, of the higher botryococcenes.     

DIVISION APPARATUS OF THE CHLOROPLAST IN NANNOCHLORIS BACILLARIS (CHLOROPHYTA)1

Chloroplast division in Nannochloris bacillaris Naumann (Chlorophyta) was examined by electron microscopy after preparation of samples by freeze-substitution. A pair of belts appeared on the surface of the outer and inner envelope membranes at the middle of the chloroplast. These belts seemed to be constructed of thin fibrils that run parallel to the longitudinal direction of the belts. The outer fibrillar belt increased in width as the constriction of the chloroplast advanced. It appears that the fibrillar belt is the division apparatus of the chloroplast. It encircles the chloroplast and finally divides the chloroplast in two as the diameter of the belt decreases.     

BEHAVIOR OF CHLOROPLAST NUCLEOIDS DURING THE CELL CYCLE OF CHLAMYDOMONAS REINHARDTII (CHLOROPHYTA) IN SYNCHRONIZED CULTURE1

Cells of Chlamydomonas reinhardtii Dangeard were synchronized under a 12:12 h light: dark regimen. They increased in size during the light period, while nuclear division, chloroplast division and cytokinesis occurred during the dark period. Zoospores were liberated toward the end of the dark period. Changes in profile and distribution of chloroplast nucleoids were followed with a fluorescence Microscope after fixation with 0.1%(w/v) glutaraldehyde followed by staining with 4′.6-diamidino-2-phenylidole (DAPI), a DNA fluorochrome. About ten granular nucleoids were dispersed in the chloroplast at the beginning of the light period (0 h). Within 4 h the nucleoids aggregated around the pyrenoid giving a compact profile. The formation of the compact aggregate of cp-nucleoids around the pyrenoid occurred with maximal frequency twice during the light period. Toward the end of the light period the nucleoids were transformed into the form of threads interconnected with fine fibrils spreading throughout the chloroplast. Initially the thread-like nucleoids fluoresced only faintly. The fluorescence of some parts of the threadlike form became brighter over a period of 6 h; these nucleoids were divided into daughter chloroplasts during chloroplast division. Soon after chloroplast division, these thread-like nucleoids were transformed into about 20 granular forms, which were gradually combined to form about ten larger granular bodies in zoospores immediately prior to liberation from mother cells. Fixation of cells with glutaraldehyde at high concentrations or treatment of cells with protease significantly modified the profiles of DAPI-stained nucleoids. The different morphologies of chloroplast nucleoids are discussed in relation to changes in configuration of their protein components.     

ABUNDANT PLASMID-LIKE DNA IN VARIOUS MEMBERS OF THE ORDERS SIPHONOCLADALES AND CLADOPHORALES (CHLOROPHYTA)1

In all seven genera belonging to the order Siphonocladales and in two genera of the Cladophorales (Chlorophyta) that were examined, extracts of total DNA contained abundant low molecular weight (LMW) molecules. This DNA typically ranged in size from 1.5 to 3.0 kb among the different genera studied. The complexity of the populations of molecules found in different genera also varied with regard to sizes and numbers of bands evident in electrophoresis gels. Our data suggest that the LMW DNA is ubiquitous among these algae since it occurred in each of our six isolates of Ernodesmis verticillata (Kützing) Børgesen collected in both the Caribbean and the Gulf of California. The LMW DNA from Ernodesmis and Ventricaria ventricosa (J. Agardh) Olsen et J. West was denser than high molecular weight (HMW) DNA in CsCl/bisbenzimide gradients. Ultrastructurally, the LMW DNA molecules were linear, averaging 0.65 μm (Ernodesmis) and 1.0 μm (Ventricaria) in length. Approximately 2%–5% of the former possessed an apparent loop or lariat at one end, as visualized in the electron microscope after rotary shadowing. The LMW DNA molecules appeared to be predominantly double-stranded DNA based upon staining with acridine orange. The 2.2-kb DNA from Ernodesmis has features typical of plasmids in that it rapidly reannealed after heat denaturation, and it was selectively enriched in an alkaline-lysis extraction procedure. In denaturing gels, this DNA migrated at >4.0 kb, indicating that the molecules may actually be single-stranded DNA species with intramolecular base pairing, each containing a long inverted repeat folded back on itself in a hairpin conformation in the native state. Digestion with mung bean nuclease revealed that additional single-stranded regions may occur in many of the LMW molecules, in addition to the hairpin loop. Although the endonuclease DNase I rapidly digested the LMW DNA, neither exonuclease III nor lambda exonuclease digested it. This suggests that the ends of these DNA molecules are protected. Collectively, the data indicate that members of the Siphonocladales and Cladophorales contain linear plasmid-like DNA molecules that appear to have a novel combination of features.     

CELL DIVISION AND THE ROLE OF THE PRIMARY WALL IN THE FILAMENTOUS DESMID ONYCHONEMA LAEVE (CHLOROPHYTA)1

Cell division and the role of the primary wall in filament formation in the desmid Onychonema laeve Nordst. were investigated by transmission and scanning electron microscopy. In addition, sequential chemical extractions and enzyme treatments were performed, on cell walls of intact filaments. Interphase cells are deeply constricted, consisting of two semicells, each elliptical in front view and circular in side view. In addition to two short lateral spines, each semicell has two apical processes that originate on opposite sides at an angle of about 15° from the central axis and overlap the adjacent cell. Division is initiated as in other desmids by a slight separation of the semicells and development of a girdle of primary wall material at the isthmus. In O. laeve the girdle of primary wall expands to form a spherical vesicle (termed a division vesicle) between the separating semicells. Nuclear division and septum formation occur in this vesicle when it is nearly the full diameter of the filament. Morphogenesis of the apical processes begins with completion of the septum, before the secondary wall appears. At maturity each apical process is surrounded by a thick layer of both secondary and primary wall, except that its capitate tip protrudes through the shroud of primary wall. Sequential treatment with hot ammonium oxalate, 4% NaOH, 17.5% NaOH and 10% chromic acid or various enzyme solutions did not cause filament breakage. SEM and TEM views of O. laeve after these treatments show intact secondary walls and intact primary wall material covering and connecting the apical processes of adjacent cells. It is the persistence of the primary wall between cells and around the apical processes that maintains the long, unbranched filamentous morphology of Onychonema laeve.     

CELL DIVISION AND FILAMENT FORMATION IN THE DESMID BAMBUSINA BREBISSONII (CHLOROPHYTA)1

Cell division and semicell expansion in the filamentous desmid Bambusina brebissonii Kütz. were investigated using transmission and scanning electron microscopy. Interphase cells are typical of desmids, containing a full complement of organelles and a cell wall penetrated by complex pores, but the cells lack a well-defined median constriction. Cell division involves an open spindle and the centripetal growth of a primary septum formed by the fusion of small, dark-staining vesicles probably derived from dictyosomes. Telophase nuclei are separated by a system of interzonal microtubules and numerous large, lighter-staining vesicles also derived from the dictyosomes. Following cell division, an elaborate replicate cross wall is formed which consists of both primary and secondary wall layers. During semicell expansion, a portion of the primary wall splits apart as the new semicells evaginate and expand to their full size. The primary wall stops splitting at a thick ring of secondary wall material leaving the cells united by the remaining common layer of primary wall. When semicell expansion is completed, the primary wall is not shed from the lateral walls of the new semicells, and pores through both primary and secondary wall layers begin to produce sheath material. However, pores in the end walls of cells do not function unless the filament is broken. The intact primary wall between cells and the absence of sheath production between cells comprise the mechanism serving to hold the cells of Bambusina brebissonii together in long filaments.     

STRUCTURE AND ABSOLUTE CONFIGURATION OF THE FLAGELLAR APPARATUS IN THE ISOGAMETES OF BATOPHORA (DASYCLADALES,CHLOROPHYTA)1

The overall appearance of the flagellar apparatus in the isogametes of Batophora oerstedii. J. Ag. is most like that which occurs in motile cells of the Ulvophyceae. Like other Ulvophyceae, the basal bodies overlap and are arranged in the 11/5 configuration, microtubular roots are arranged in a cruciate pattern and system II striated fibers are present. The basal body connective which generally lacks striation in the Ulvophyceae is clearly different in Batophora, being composed of two large non-striated halves which connect to the anterior surface of each basal body and are then connected to one another by a distinctly fibrous centrally striated region. This variation in the basal body connective and the presence of two posteriorly directed system II striated fibers is clearly different from homologous structures reported in siphonous green algae of the Caulerpales. Based upon these variations and similarities among flagellar apparatus components in siphonous green algae, it is suggested that the Dasycladales and Siphonodadales are more closely related to one another than to the Caulerpales.     

GROWTH CHARACTERISTICS OF MARINE PHYTOPLANKTON DETERMINED BY CELL CYCLE PROTEINS: THE CELL CYCLE OF ETHMODISCUS REX (BACILLARIOPHYCEAE) IN THE SOUTHWESTERN NORTH ATLANTIC OCEAN AND CARIBBEAN SEA1

Ethmodiscus spp. is an important contributor to oceanic tropical-ooze sediments and thus might be an important transport vehicle of carbon from the ocean surface to sediments. The knowledge of its cell cycle and growth rate, which is still lacking, is necessary to evaluate the importance of Ethmodiscus in nutrient cycling and to solve the discrepancy between its high sedimentary abundance and rarity in the plankton. We used immunofluorescence of a cell cycle protein, prolqerating cell nuclear antigen (PCNA), and DNA-specific staining to study the progression of the cell cycle and roughly estimate the growth rate for E. rex (Rattray) Wiseman and Hendey in the southwestern North Atlantic Ocean and Caribbean Sea in June 1994 and January 1995. During the cell division cycle, the chloroplasts appeared to synthesize DNA before the nucleus (S phase). Following the S phase, the nucleus moved from one end of the cell toward the center underneath the midline of the girdle band (G2 phase) where it divided (M phase). During a very brief period, the parent cell split and moved apart from the girdle midline, and two new valves were produced (late M phase). The two daughter nuclei apparently remained attached at the joint of the two newly produced valves, where they appeared to be responsible for coordinating the symmetrical formation of the new valves. The morphologically complete daughter cells remained joined for a short period of time before separating into solitary cells whose nucleus was located at one end of the cell. Derived from the phase fraction curves, the duration of the cell cycle phases decreased in the order from G1, S, G2, to M. A conservative estimate of the growth rate in the study area obtained by using PCNA immunostaining was 0.39–0.46 d^?1 in June and 0.15 d^?1 in January. The validity and implication of the growth rate estimates are discussed.     

COMPARATIVE PHYSIOLOGY AND BIOCHEMISTRY AND TAXONOMIC ASSIGNMENT OF THE CHLORELLA (CHLOROPHYCEAE) STRAINS OF THE CULTURE COLLECTION OF THE UNIVERSITY OF TEXAS AT AUSTIN1

The species of the genus Chlorella exhibit considerable biochemical and physiological differences. Therefore, it is important to select for and utilize in research or biotechnology correctly identified strains of the species having the most favorable properties for the respective project. We examined the Chlorella strains of the University of Texas collection at Austin, Texas, according to species-specific chemotaxonomic characters and assigned 58 strains to 10 well-established species (only 17 of these strains were correctly named before).     

SUGAR COMPOSITION OF THE CELL WALL AND THE TAXONOMY OF CHLORELLA (CHLOROPHYCEAE)1

Cell walls of forty Chlorella strains covering all species of the Algal Collection of Göttingen (C. fusca var. vacuolata, C. kessleri, C. luteoviridis, C. minutissima, C. protothecoides, C. saccharophila, C. sorokiniana, C. vulgaris, and C. zofingiensis) were compared. The nine species were divided into two groups according to the major sugar in the rigid wall. The first group had a glucose-mannose-rigid wall and included C. fusca var. vacuolata, C. luteoviridis, C. minutissima, C. protothecoides, C. saccharophila, and C. zofingiensis. The second group, with a glucosamine-rigid wall, included C. kessleri, C. sorokiniana, and C. vulgaris. Chlorella strains of the nine species were further classified by constituent sugars, ruthenium red stainability, and anisotropy of the cell walls.     

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.     

THE OPTIMUM PH OF FOUR STRAINS OF ACIDOPHILIC SNOW ALGAE IN THE GENUS CHLOROMONAS (CHLOROPHYTA) AND POSSIBLE EFFECTS OF ACID PRECIPITATION1

Four axenic strains of snow algae were examined for optimum pH under laboratory conditions using M-1 growth medium. Growth was measured using cell counts, cell measurements and absorbance readings at 440 nm. Strains C204 and C479A of Chloromonas sp. from the Adirondack Mountains, New York, grew optimally at pH 4.0 to 5.0. Strains C381F and C381G, Chloromonas polyptera (Fritsch) Hoh., Mull. & Roem. from the White Mountains, Arizona, grew optimally at pH 4.5 to 5.0. Growth was significantly higher at pH 4.0 in the northeastern species (Chloromonas sp.), but no significant difference was observed in final growth at pH 4.5, 5.0 and 5.5 between species. It is postulated that the more acidic precipitation in the northeastern United States may be selecting for strains of snow algae with greater tolerance to acidity than in strains from the southwestern United States or that the different pH optima reported are simply species differences. New York strain C204 was also grown in heavily buffered AM medium where it had an optimum pH of 5.0, but cells became irregularly shaped and tended to clump at pH 6.0 to 7.0. Growth of C204 in AM medium was significantly lower than in M-1 medium for snow algae. These findings justify the use of M-1 medium for this type of experimentation.