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.     

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.     

METHYLAMINE UPTAKE IN THE GREEN ALGA CHLORELLA PYRENOIDOSA1

Methylamine uptake in nitrogen-starved Chlorella pyrenoidosa Beij. follows Michaelis-Menten kinetics: maximum uptake is about 1.6 nmol μl^?1· cells · min^?1, half-saturation occurs at 4 μM methylamine, and the slope in the range where uptake is proportional to concentration is 0.4 nmol μl^?1· min^?1·μM^?1. In cells grown in the presence of a non-limiting nitrogen concentration, methylamine uptake is directly proportional to concentration up to at least 0.5 mM, and the slope is 1/500 that for starved cells. Similar uptake kinetics have been reported for Penicillium chrysogenum and attributed to an inducible “ammonium permease.” Apparently, a similar permease occurs in algae.     

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.     

A LARGE SCALE QUASI-CRYSTALLINE LAMELLAR LATTICE IN CHLOROPLASTS OF THE GREEN ALGA ZYGNEMA

A quasi-crystalline lamellar lattice was observed in chloroplasts of the filamentous green alga Zygnema. The lattice does not appear in the cells until cultures are at the end of the log phase of growth. Pseudograna are also present and become more numerous towards the middle of the log phase. The three-dimensional lattice superficially resembles the configuration of cubic prolamellar bodies but is about 10 times larger and is entirely different in internal structure. The lattice is composed of one or two appressed thylakoids in a stroma matrix which is bounded on each side by a single thylakoid membrane. This multilayered sandwich of membranes and matrix occupies a position equivalent to the single membrane of a cubic prolamellar body.     

TRACING SPECIES INVASION IN CODIUM,A SIPHONOUS GREEN ALGA,USING MOLECULAR TOOLS

The siphonous green alga Codium fragile occurs in many temperate marine regions and is composed of a number of distinct subspecies. Included in this taxon is the common open coast C. fragile subsp. fragile of the northeast Pacific and the weedy C. fragile subsp. tomentosoides which has invaded temperate marine communities in the Atlantic and Pacific oceans. The center of origin of this weedy subspecies is not known, although it is thought to have dispersed from the northwest Pacific. To examine the relationship of the weedy subspecies to the indigenous northeast Pacific form, chloroplast DNA was compared. Each of these subspecies has a restriction map that is uniform throughout its geographic distribution, and the patterns are distinct from each other and from other Codium species examined. However, the two share an almost identical genome size and arrangement of genes. A population in San Francisco Bay was found to be indistinguishable from the weed C. fragile subsp. tomentosoides from the Atlantic. The potential for using molecular data in solving systematic problems in Codium has been clearly demonstrated.     

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.     

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.     

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.     

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.     

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.     

A CLASSROOM DEMONSTRATION OF ZOOSPORE PRODUCTION IN THE GREEN ALGA SCHIZOMERIS LEIBLEINII KUETZ.12

Using the parenchymatous ulotrichalean green alga Schizomeris leibleinii, a dependable technique for the classroom demonstration of zoospore production is described. The onset of zoospore release can be predicted accurately by controlling light, temperature, and medium. Large numbers of zoospores are released.     

SPONTANEOUS MUTATION ACCUMULATION IN MULTIPLE STRAINS OF THE GREEN ALGA,CHLAMYDOMONAS REINHARDTII

Estimates of mutational parameters, such as the average fitness effect of a new mutation and the rate at which new genetic variation for fitness is created by mutation, are important for the understanding of many biological processes. However, the causes of interspecific variation in mutational parameters and the extent to which they vary within species remain largely unknown. We maintained multiple strains of the unicellular eukaryote Chlamydomonas reinhardtii, for approximately 1000 generations under relaxed selection by transferring a single cell every ～10 generations. Mean fitness of the lines tended to decline with generations of mutation accumulation whereas mutational variance increased. We did not find any evidence for differences among strains in any of the mutational parameters estimated. The overall change in mean fitness per cell division and rate of input of mutational variance per cell division were more similar to values observed in multicellular organisms than to those in other single‐celled microbes. However, after taking into account differences in genome size among species, estimates from multicellular organisms and microbes, including our new estimates from C. reinhardtii, become substantially more similar. Thus, we suggest that variation in genome size is an important determinant of interspecific variation in mutational parameters.     

MITOSIS,CYTOKINESIS, AND COLONY FORMATION IN THE GREEN ALGA SORASTRUM1

Synchronous mitotic divisions produce multi-nucleate cells of Sorastrum. Perinuclear envelopes of endoplasmic reticulum and a virtually intact nuclear envelope enclose mitotic nuclei. Cytoplasmic cleavage, which shirts before the last round of Synchronous mitoses, gives rise to uninucleate fragments which differentiate to form zoospores. These zoospores are released into a spherical vesicle, presumably derived from the inner layer of the parental cell wall, in which they swarm actively before aggregating as a spherical colony. The roughly conical shaped zoospores apparently adhere laterally before withdrawing their flagella and extending horns and a stipe, which, following wall deposition, interconnects the cells at the center of the colony. The probable role of the microtubules, which underlie the plasmalemma of aggregating cells, in determining the shape of both the cells and the colony itself is discussed.     

THE ROLE OF ZINC FINGER PROTEIN IN RNAi INTERFERENCE IN A UNICELLULAR GREEN ALGA CHLAMYDOMONAS REINHARDTII (CHLOROPHYCEAE)

In our previous study, we generated a strain of 19‐P (1030) in which artificial RNA interference (RNAi) was induced by transcribing a hairpin RNA of ~780‐bp stem. We utilized this RNAi‐induced strain to uncover RNAi‐related genes. Random insertional mutagenesis was performed to generate tag‐mutants that show a RNAi deficient phenotype. The 92‐12C is one such tag‐mutant, which bears a 14‐kb deletion in chromosome 1. Complementation of 92‐12C revealed that a protein gene, including a Cys‐Cys‐Cys‐His‐type zinc finger motif and an ankyrin repeat motif, is essential for effective RNAi in Chlamydomonas reinhardtii (Dangeard). BLAST analysis revealed that the zinc finger protein is homologous to an mRNA splicing‐related protein of other species. Therefore, one of the probable scenarios is that mRNA coding for RNAi‐related proteins cannot be properly spliced, which causes RNAi deficiency in the 92‐12C tag‐mutant.     

丝状固氮蓝藻柱胞鱼腥藻原生质球的培养再生

丝状固氮蓝藻柱胞鱼腥藻 (Anabaena cylindrica)在含 0 .1mol/ L KCl的 Allen液体培养基中培养7～ 9d,90 %以上细胞转化为原生质球或半原生质球 ,对低渗敏感或抗低渗。此种材料经 0 .1%溶菌酶 ,在 2 8℃下处理 3～ 4h,细胞低渗破裂率约 10 0 % ,成为质量较高的原生质球。用 0 .15mol/ L Ca Cl2 液体无机盐培养基培养 ,原生质球再生和分裂。不同原生质球再生不同步 ,最快培养 3 d分裂。再生分裂的主要方式为均等分裂 ,但有不规则分裂和出芽方式 ,再生率在 2 5%以上。     

一株耐碳酸氢钠产油绿藻的絮凝收集

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