NUTRITION OF THE GREEN ALGA GOLENKINIA

Defined media were established for four strains of Golenkinia. Strains 929 and 930 require thiamine and vitamin B₁₂, strain 931 requires the latter and is stimulated by thiamine; only strain 320 and a mutant of strain 931 are capable of completely autotrophic growth. The optimal nitrogen concentration is 0.025 m except for strain 931 where it is 0.015 m and the requirement may be met by nitrate, ammonium, or urea. Optimal levels of the other components are: 0.0025 m KH₂PO₄, 0.0005 to 0.001 m MgSO₄, and 50 ml of a stock trace-element solution per liter. Only strain 931 required calcium; however, its addition stimulates the growth of strains 929 and 930. The optimal pH before autoclaving is 6.8. Maximal growth rates of two to three cell doublings per day and yields of 26 to 30 (log₂) cells per ml were obtained. These growth data compare favorably with those for other unicellular green algae.     

A POSSIBLE VIRUS INFECTION IN THE GREEN ALGA OEDOGONIUM1

Developing germlings of the green alga Oedogonium have been sectioned and some were found to contain particles presumed to be those of a virus. Fully formed particles were hexagonal in a 2-dimensional section, about 240 nm in diameter with a densely stained surface layer and core; what appear to be different stages in their assembly were also seen. Many of the particles were clustered together in groups surrounded by aggregations of ribosomes. In cells containing many of the particles, the germlings' cytoplasm and organelles showed considerable evidence of disruption and breakdown. These particles have only been detected in germlings, so infection by this presumed virus was probably initiated in the naked, motile zoospore.     

THE EFFECTS OF ISOPROPYL N-PHENYL CARBAMATE ON THE GREEN ALGA OEDOGONIUM CARDIACUM : I. Cell Division

Cell division in vegetative filaments of the green alga Oedogonium cardiacum is presented as an experimental system. We report on how we have used this system to study the effects of isopropyl N-phenylcarbamate (IPC) on the mitotic apparatus and on the phycoplast, a planar array of cytokinetic microtubules. Polymerization of microtubules was prevented when filaments, synchronized by a light/dark regime and chilled (2°C) while in metaphase or just before phycoplast formation, were exposed to 5.5 x 10^-4 M IPC and then returned to room temperature. Spindles reformed or phycoplasts formed when these filaments were transferred to growth medium free of IPC. However, the orientation of both microtubular systems was disturbed: the mitotic apparatus often contained three poles, frequently forming three daughter nuclei upon karyokinesis; the phycoplast was often stellate rather than planar, and it sometimes was displaced to the side of both daughter nuclei, resulting in a binucleate and an anucleate cell upon cytokinesis. Our results suggest that IPC (a) prevents the assembly of microtubules, (b) increases the number of functional polar bodies, and (c) affects the orientation of microtubules in O. cardiacum. High voltage (1,000 kV) electron microscopy of 0.5-µm thick sections allowed us to visualize the polar structures, which were not discernible in thin sections.     

DIFFERENTIATION OF THE GREEN ALGA CODIUM FRAGILE

Clonal cultures of Codium fragile were established from both swimming cells and vegetative filaments. In the laboratory axis primordia differentiate from heterotrichous juveniles only when cultures are agitated on a reciprocating shaker. The shear forces created by mechanical agitation are essential both for initiation and maintenance of primordia. Contact guidance of growing coenocytic filaments indicates mutual adhesion of filaments as the basis for the differentiation process.     

CELL WALL CARBOHYDRATE EPITOPES IN THE GREEN ALGA OEDOGONIUM BHARUCHAE F. MINOR (OEDOGONIALES,CHLOROPHYTA)1

Cell wall changes in vegetative and suffultory cells (SCs) and in oogonial structures from Oedogonium bharuchae N. D. Kamat f. minor Vélez were characterized using monoclonal antibodies against several carbohydrate epitopes. Vegetative cells and SCs develop only a primary cell wall (PCW), whereas mature oogonial cells secrete a second wall, the oogonium cell wall (OCW). Based on histochemical and immunolabeling results, (1→4)‐β‐glucans in the form of crystalline cellulose together with a variable degree of Me‐esterified homogalacturonans (HGs) and hydroxyproline‐rich glycoprotein (HRGP) epitopes were detected in the PCW. The OCW showed arabinosides of the extensin type and low levels of arabinogalactan‐protein (AGP) glycans but lacked cellulose, at least in its crystalline form. Surprisingly, strong colabeling in the cytoplasm of mature oogonia cells with three different antibodies (LM‐5, LM‐6, and CCRC‐M2) was found, suggesting the presence of rhamnogalacturonan I (RG‐I)–like structures. Our results are discussed relating the possible functions of these cell wall epitopes with polysaccharides and O‐glycoproteins during oogonium differentiation. This study represents the first attempt to characterize these two types of cell walls in O. bharuchae, comparing their similarities and differences with those from other green algae and land plants. This work represents a contribution to the understanding of how cell walls have evolved from simple few‐celled to complex multicelled organisms.     

CYTOLOGICAL STUDIES OF OEDOGONIUM. II. CHROMOSOME NUMBERS IN TWELVE SPECIES OF OEDOGONIUM

Mitotic chromosome counts are reported for 12 species of Oedogonium with the following distribution: two species with 13 chromosomes, two species with 16 chromosomes, five species with 17 chromosomes one species with 19 chromosomes, one species with 32 chromosomes, and one species with 38 + 1 chromosomes. Diploid strains of two species are illustrated and discussed. Cytological comparison of species establishes that there is great diversity in Oedogonium with relation to chromosome number, size, and morphology.     

ULTRASTRUCTURE OF MITOSIS AND CYTOKINESIS IN THE MULTINUCLEATE GREEN ALGA ACROSIPHONIA

The processes of mitosis and cytokinesis in the multinucleate green alga Acrosiphonia have been examined in the light and electron microscopes. The course of events in division includes thickening of the chloroplast and migration of numerous nuclei and other cytoplasmic incusions to form a band in which mitosis occurs, while other nuclei in the same cell but not in the band do not divide. Centrioles and microtubules are associated with migrated and dividing nuclei but not with nonmigrated, nondividing nuclei. Cytokinesis is accomplished in the region of the band, by means of an annular furrow which is preceded by a hoop of microtubules. No other microtubules are associated with the furrow. Characteristics of nuclear and cell division in Acrosiphonia are compared with those of other multinucleate cells and with those of other green algae.     

ULTRASTRUCTURAL ORGANIZATION OF CHLOROPLAST THYLAKOIDS OF THE GREEN ALGA "OOCYSTIS MARSSONII"

Intact cells of "Oocystis marssonii" were thin sectioned and freeze-etched, using conventional and double-recovery techniques. Thylakoids extend the length of the single chloroplast and occur in stacks of three to five. The peripheral thylakoids in a stack often alternate between adjacent stacks. Interpretation of double-recovery results suggests that membranes in unstacked regions are asymmetrical, with one face smooth and the matching face covered with closely packed 85–90 Å diameter particles. Adjacent membranes in stacked regions evidently share 170 Å diameter particles, and either membrane in a stacked region may fracture. The two fracture planes thus made possible may expose nearly entire 170 Å particles or only the upper portion of such particles, creating in the latter case images of 125–135 Å diameter particles. Fracture planes in all cases appear to occur through the interior of the membrane, in the plane between the hydrophobic ends of the lipid bilayer proposed in numerous membrane models.     

FLAGELLAR APPARATUS OF THE BIFLAGELLATE ZOOSPORES OF THE ENIGMATIC MARINE GREEN ALGA BLASTOPHYSA RHIZOPUS1

The flagellar apparatus of the biflagellate zoospores from Blastophysa rhizopus Reinke has 180° rotational symmetry of the major components and counterclockwise absolute orientation. The basal bodies are connected anteriorly by a prominent striated distal fiber and posteriorly by two proximal striated bands. The C microtubules in the basal bodies terminate proximal to the transition region. Terminal caps and well-defined proximal sheaths are absent. The four microtubular rootlets diverge at a very small angle from the basal bodies. Six to eight (usually seven) microtubules are present in the s rootlets and two microtubules in the d rootlets. Rootlet 1s is associated with the eyespot. Each d rootlet is subtended by a coarsely striated fiber. Rootlet Id also has a finely striated fiber, roughly opposite the coarsely striated fiber, associated with it. Rhizoplasts and mating structures were not observed. Ultrastructural features of B. rhizopus zoospores are essentially identical with those found in examined members of the Siphonocladales sensu lato (= Siphonocladadales/Cladophorales complex) and Dasycladales, and have relatively few features in common with motile cells of caulerpalean algae. Blastophysa rhizopus probably does not represent an intermediate between the Siphonocladadales and the Caulerpales. Its evolutionary history is different from that of other algae placed in the siphonocladalean family Chaetosiphonaceae. Whether or not Blastophysa is representative of the ancestor to the Siphonodadales and Dasycladales is unclear.     

CURRENT STATUS OF THE INVASIVE GREEN ALGA CODIUM FRAGILE IN EASTERN CANADA

Ten years after the initial discovery of Codium fragile in eastern Canada on the Atlantic coast of Nova Scotia, C. fragile has extended its range considerably to northern Nova Scotia, southern New Brunswick and Prince Edward Island (P.E.I.) in the Gulf of St. Lawrence. In all principal areas C. fragile subsp. tomentosoides is present. In two regions of Prince Edward Island, Codium is becoming extremely abundant, and is known by local fishers as the "oyster thief". A potential second taxon of Codium was collected in 1999 at three sites in Malpeque Bay where it is locally abundant. These plants are distinguished from C. fragile subsp. tomentosoides by their smaller frond size, flatter utricle tops, smaller mucrons, and smaller gametangia. Principal component analysis distinguishes plants of the two types. The abundance of C. fragile on both hard bottom and soft bottom substrata suggests that it will become the dominant alga in many subtidal algal communities in eastern Canada.     

GROWTH OF THE GREEN ALGA CODIUM FRAGILE IN A CONNECTICUT ESTUARY1

An in situ comparison of environmental and physiological factors was undertaken in 1971–72 (15 months) in the Niantic River estuary to elucidate some of the important aspects, of the growth and development of the seaweed Codium fragile. In general, Codium in the estuary has a growing season of from 6 to 9 months. Growth increments during this period are relatively constant. Temperature and salinity are the main limiting factors to growth, although low summer concentrations of inorganic nitrogen may also be involved. Reproduction by means of swarmers occurs only in late summer or autumn. Codium appears highly adapted to the role of a colonizing species. It not only possesses the capability to occupy the harshest of environments, but also a system of reproductive alternatives which facilitates rapid colonization. The availability of substrate for attachment is the chief factor limiting its spread in this estuarine system.     

FINE STRUCTURE OF CAPITULAR FILAMENTS IN THE COENOCYTIC GREEN ALGA PENICILLUS1,2,3

Capitular filaments of Penicillus capitatus contain a large central vacuole. The parietal cytoplasm is densely packed, devoid of chloroplasts in the growing tip, and becomes convoluted and sponge-like as extensions of the vacuole penetrate the cytoplasm in mature portions of the filament. Structure of organelles and their distribution in the filament are described. The vacuole contains a variety of inclusions, such as membranous configurations, spherical bodies, electron dense bodies, and calcium oxalate monohydrate crystals, each of the latter surrounded by a chamber associated with microtubules. Endophytic bacteria are present throughout the vacuole and occasionally in the tip cytoplasm. Some vacuolar components of P. pyriformis are described for comparison.     

THE GLYCEROL PERMEABILITY OF THE PLASMALEMMA OF THE HALOTOLERANT GREEN ALGA DUNALIELLA PARVA (VOLVOCALES)1

The glycerol permeability of the plasmalemma of the green alga Dunaliella parva Lerche was investigated by efflux studies with labelled glycerol, by enzymatic determination of glycerol leakage, and the determination of the reflection coefficient from osmotically induced volume changes (zero flow method). All results indicate that the plasmalemma of D. parva does not exhibit a special low permeability towards glycerol as would be expected from a glycerol accumulating alga. Rather, significant amounts of glycerol diffuse continuously into the medium following the glycerol concentration gradient between the cells and the medium. Efflux rates vary between 0.1 and 2 μmoles glycerol·mg^?1 chlorophyll·h^?1 depending on the external NaCl concentration. After one day up to 25% of the total glycerol of the algal suspension was found in the medium. Within 10 days this value can increase to 60%, depending on the growth constant of the culture. The reflection coefficient σ was determined to be 0.87, the permeability coefficient 2800 × 10^?11 m·sec^?1. To maintain a proper endogenous glycerol level corresponding to the external osmotic pressure, glycerol efflux in D. parva has to be balanced by a continuous synthesis of glycerol. D. parva follows the strategy of “glycerol efflux tolerance” instead of “glycerol efflux avoidance”. The alga has to pay the energetic costs of this strategy of tolerance.     

EXPERIMENTAL EVOLUTION OF HETEROTROPHY IN A GREEN ALGA

Laboratory populations of a green alga cultured in the dark with an organic substrate evolved into efficient heterotrophs with faster growth, higher fitness, and increased responsiveness to substrate concentration. Their phenotypes were almost entirely attributable to selection, rather than to history or ancestry. The fitness of evolved lines in the light was uniformly depressed, presumably through the accumulation of conditionally deleterious genes governing photosynthesis. Some evolved lines were no longer able to grow in the light and are thereby permanently isolated from their ancestors. Specialized autotrophs and heterotrophs may often evolve in algae through long‐term shifts in the conditions of growth.