Reconsideration of the taxonomy of ellipsoidal species of Chlorella (Trebouxiophyceae, Chlorophyta), with establishment of Watanabea sen. nov.

Subcultures of SAG 211–9b and 1AM C-211, ultimately derived from CCAP 211/9b, a strain isolated by Pringsheim in 1939 and identified as Chlorella sac-charophila (Kruger) Migula were observed using light and electron microscopy. Their morphology proved to be basically identical. Both have two forms of cells, one (E-form) narrowly to broadly ellipsoidal, the other (S-form) ovoid to spheroidal. The cell wall of both forms is composed of a single smooth layer. The chloroplast of young cells is trough-like or saucer-shaped with a smooth margin, while that of mature cells is band- or cup-shaped with deep incisions. The thylakoid lamellae are loosely stacked and neither form has a pyrenoid. Both types of cells are capable of producing autospores: eight to 16 in E-form cells, two to four in S-form cells. These morphological features are different from those of C. saccharophila, which has a pyrenoid and produces only one form of autospores. In the absence of any existing genus that includes Chlorella-like algae with a simple cell wall, no pyrenoid, and two forms of mature cells and autospores, a new genus, Watanabea, is proposed with the type species W. reniformis.     

Circumscription of the order Ectocarpales (Phaeophyceae): bibliographical synthesis and molecular evidence

Phylogenetic analyses of 27 brown algae including the type genera of the orders Chordariales, Dictyosiphonales, Ectocarpales sensu stricto, and Scytosiphonales, using partial SSU + LSU combined rDNA sequence data, supports a broadly circumscribed order Ectocarpales. This order is redefined to include taxa possessing an exserted, pedunculated pyrenoid. Previous then taxa were placed in the Ectocarpales sensu stricto, the Chordariales, the Dictyosiphonales, or the Scytosiphonales. Algae either lacking a pyrenoid, and sometimes included in the Ectocarpales (Tilopteridales, Ralfsiales sensu Nakamura), or which possess a non-pedunculated pyrenoid (such as those placed in the recently proposed order Scytothamnales, as well as Asteronema, Asterocladon and Bachelotia), are excluded from the Ectocarpales.     

OBSERVATIONS ON “VAMPYRELLA PENULA-STYLODINIUM SPHAERA” AND THE ULTRASTRUCTURE OF THE REPRODUCTIVE CYST

“Vampyrella-Stylodinium,” an artificial name for a predaceous organism of uncertain taxonomic position, has at least three distinct phases in its life history: the amoeboid phase, both free-floating and attached; the feeding cyst or immobile phase; and flagellated gymnodinoid swarmers. The orange free-floating amoeba has unbranched, filose pseudopodia and several contractile vacuoles. When feeding on the filamentous green alga Oedogonium, the pseudopodia shorten and rearrange. After dissolution of part of the Oedogonium cell wall, the amoeba ingests the host protoplast. Then a stalked reproductive cyst may form. This cyst changes color from green to light orange as it matures. At the time of excystment, the cyst has a smooth outer wall, a spinose inner wall, and a well-delineated phagocytic vacuole. As this vacuole moves from its central position to the cyst's periphery, the walls rupture and 2-4 amoebulae emerge. With TEM observations, the reproductive cyst is shown to be multinucleate. Each nucleus is eukaryotic in organization and possesses one nucleolus. Mitochondria have tubular cristae and no structures unique to the division Pyrrhophyta are observed. Although this stage of the life history does not have a dinokaryotic nucleus, the gymnodinoid swarmers that can emerge from the reproductive cyst, do. Like other parasites which have been assigned to the division Pyrrhophyta, “Vampyrella-Stylodinium” does not conform well to the generalized concept of a dinoflagellate.     

Ultrastructural morphology of the reproductive swarmers of Sphaerozoum punctatum (Huxley) from the East China Sea

Reproductive swarmers of the polycystine radiolarian Sphaerozoum punctatum (Huxley) collected from the East China Sea were examined using light, scanning and transmission electron microscopy. The swarmer cells were about 8–10 μm in length with a pear-like shape and a conical end with two flagella. A nucleus, mitochondria, Golgi body, lipid droplets and, characteristically, a single, large, vacuole-bound SrSO₄ crystal were present in the cytoplasm. Centering on the crystal inclusion, swarmers swam in a rapid rotational movement both clockwise and anticlockwise. Small subunit (SSU) rDNA sequences obtained for the reproductive swarmer cells from S. punctatum show a monophyletic group together with colonial spumellarians and grouped with S. punctatum from Bermuda in the clade. The morphological features and molecular phylogeny of the reproductive swarmers of S. punctatum show evidence of ancestral traits of radiolarians; acantharians and polycystines have a common ancestry. In addition, SrSO₄ inclusion of the swarmer cell may be a form of ballast deposited by the swarmer to allow proper positioning in the water column. We hypothesize that radiolarian-affiliated sequences from SSU rDNA clone libraries of marine picoeukaryotes may be derived from the picoplanktonic cells of radiolarians; i.e., small flagellated life stages such as reproductive swarmers or gametes.     

Generic and species concepts in Microglena (previously the Chlamydomonas monadina group) revised using an integrative approach

Traditionally the genus Microglena Ehrenberg has been used to contain species that belong to the Chrysophyceae; however, the type species of Microglena, M. monadina, represents a green alga, which was later transferred to the genus Chlamydomonas. The taxonomic status of the genus has therefore remained unclear. We investigated 15 strains previously assigned to C. monadina and two marine species (C. reginae and C. uva-maris) using an integrative approach. Phylogenetic analyses of SSU and ITS rDNA sequences revealed that all strains form a monophyletic lineage within the Chlorophyceae containing species from different habitats. The strains studied showed similar morphology with respect to cell shape and size, but showed differences in chloroplast and pyrenoid structures. Some representatives of this group have the same type of sexual reproduction (homothallic advanced anisogamy). Three different morphotypes could be recognized. Strains belonging to type I have a cup-shaped chloroplast with a massive basal part, in which a large, single, ellipsoidal pyrenoid is located. The members of type II also have a cup-shaped chloroplast, which is partly lobed and has a thinner basal part than type I; here the pyrenoid is half-ring or horseshoe-shaped and occupies different positions in the chloroplast depending on the strain. The strains of type III have multiple pyrenoids, which appear to have developed from the subdivision of a single ring-shaped pyrenoid into several parts. We compared the results of our morphological investigations with the literature and found that 15 strains could be identified with existing species. Two strains did not fit with any described species. As a result of our study, we transfer all strains to the genus Microglena, propose 11 new combinations, and describe two new species. Comparison of the ITS-1 and ITS-2 secondary structures confirmed the species delineations. All species have characteristic compensatory base changes in their ITS secondary structures and are supported by ITS-2 DNA barcodes.     

Studies on Kuckuckia spinosa (Fucophyceae, Sorocarpaceae): life history, temperature gradient experiments, and synonymy

Kuckuckia spinosa and Kuckuckia kylinii are distinguished mainly by the presence of basally sheathed true hairs in the latter. However, culture studies have shown that the sheath is an ephemeral structure as it splits, breaks off, and the basal remnant disintegrates and disappears eventually. Therefore, Kuckuckia kylinii gradually develops into Kuckuckia spinosa , also with regard to filament diameter. Consequently, the two species are merged with priority to K. spinosa. The life history is of the direct type as the swarmers from the plurilocular sporangia develop by immediate differentiation into plants similar to the mother plant. It has been confirmed experimentally that this species belongs to the warm temperate Mediterranean-Atlantic group with a southern reproductive boundary close to 25°C and a northern reproductive boundary close to the 10°C summer isotherms. This is the first report of this species from the Canary Islands.     

中国淡水绿藻纲新记录属麦可属(Mychonastes)

从我国滇池分离、培养获得2株超微真核藻, 对其进行形态学、细胞超微结构和18S rRNA基因序列分析。结果表明: 藻株在细胞形态、结构和繁殖方式具麦可属(Mychonastes Simpson Van Valkenburg)特征, 细胞壁2层, 外层细胞壁表面具不规则肋网和典型的暗-明-暗结构; 具备一套简单的细胞器, 包括细胞核、不具蛋白核的叶绿体和线粒体各1个, 叶绿体周生、杯状, 占据细胞大部分体积; 以似亲孢子方式繁殖。结合18S rRNA序列分析, 将其归为麦可属, 属于绿藻纲、绿藻门, 是该属在我国淡水湖泊的首次描述。       

ULTRASTRUCTURE OF GLOEODINIUM MONTANUM (DINOPHYCEAE)1

The freshwater dinoflagellate Gloeodinium montanum Klebs (1912) was examined with transmission and scanning electron microscopy. Micrographs of ultrathin sections revealed a series of membrane layers rather than the usual dinoflagellate theca in vegetative cysts and in legates. Swarmers had distinct pellicles but appeared to be devoid of thecal plates and vesicles. The organization of cysts and swarmers appeared remarkably similar. All cell types had typical dinoflagellate nuclei with condensed chromosomes. Chloraplasts had girdle lamellae. One pyrenoid per cell was also present in chloroplasts of vegetative cysts. Starch grains and oil globules were distributed throughout the cytoplasm. Large accumulation bodies and polyvesicular vacuoles were found in aging cysts. Trichocysts and flagellar hairs were absent. Two types of intra-cellular prokaryotic organisms were discovered.     

Life cycle,behavior and morphology of a new tide pool dinoflagellate,Gymnodinium pyrenoidosum sp. nov. (Gymnodiniales,Pyrrhophyta)

A new tide pool dinoflagellate,Gymnodinium pyrenoidosum Horiguchi et Chihara sp. nov. is described from central Japan. It was found to form dense blooms with a characteristic greenish color from April to November. The species exhibits a characteristic diurnal vertical migration and an alternation of a motile with a nonmotile phase, which are dependent on light intensity and tidal movement. Cells of the motile phase are unarmored and relatively small. They have a single, reticulate chloroplast, orange stigma situated near the sulcus and conspicuous pyrenoid in epicone. The alga reproduces itself by means of zoospores which are produced by the bipartition of protoplasm within the parent cell wall during the nonmotile stage which occurs at night. The occurrence of another type of motile cell, termed a macroswarmer, which differs from normal zoospore in size and shape has also been demonstrated.     

Ultrastructure of Characiochloris acuminata Lee et Bold

The ultrastructure of the vegetative cell and zoospore of Characiochloris acuminata Lee et Bold (Chlorangiellaceae, Tetrasporales, Chlorophyceae) is described.

The vegetative cell is distinctive in having numerous contractile vacuoles which are randomly distributed in the cytoplasm and visible through the fissures of the parietal chloroplast. A single pyrenoid, embedded in the chloroplast, is penetrated by cytoplasmic canals which are lined by the chloroplast envelope. The vegetative cell is attached to the substrate or host by two flagellar remnants (retained from the zoospore stage), each of which is ensheathed in a gelatinous tube through the cell wall at the cell base. The basal bodies are apparently abscissed from the flagellar shaft by a unit membrane which becomes continuous with the plasma membrane.

The zoospore is biflagellate, with the flagella equal in length, smooth and longer than the cell body. The flagellar sheath is characteristically undulate and the two flagellar bases are connected by a dense interflagellar fibre. The large nucleus has a conspicuously inflated nuclear envelope and the pyrenoid is similar to that of the vegetative cell.     

Observations with the electron microscope on Asteromonas gracilis Artari emend. (Stephanoptera gracilis (Artari) wisl.), with some comparative observations on Dunaliella Sp

A clone of Asteromonas gracilis has been examined in culture by means of light and electron microscopy. The morphological observations of Artari (1913) have been confirmed and extended. New observations include demonstration of an unusual type of pyrenoid somewhat resembling that of Prasinocladus, with tubular channels entering the pyrenoid centre from outside the plastid and penetrated by haustorial outgrowths from the nuclear envelope. The very different structure characteristic of Dunaliella is briefly illustrated. Other comparisons indicate that A. gracilis is the type species and sole known representative of a very distinct genus which cannot be confused with Pyramimonas and is unlikely to be synonymous with the imperfectly known genus, Stephanoptera.     

Arrays of Granules Associated with the Plasmalemma in Swarmers of Cladophora

The surface region of swarmers of the green alga, Cladophorarupestris, has been studied in both chemically fixed and freeze-etchedmaterial. Swarmers prepared by each of these methods show arraysof granules outside the plasmalemma, covered by a layer of fibrousmaterial of unknown composition which completely surrounds thecell. In the case of freeze-etched swarmers, the granules areformed into a lattice closely similar to that predicted earlierin the ‘ordered granule hypothesis’ of cellulosebiosynthesis. No microfibrils have yet been observed in associationwith the granules, but their location in the region of wallformation, at a time when this process is in its early stages,lends support to the possibility that the granules may be theorganelles responsible for the production and orientation ofthe cellulose microfibrillar component of the cell wall. Noevidence was observed during this work to suggest that any otherorganelle is responsible for cellulose formation.     

Observations on the ultrastructure of representatives of the genera Hemiselmis and Chroomonas (Cryptophyceae)

Hemiselmis rufescens Parke and three species of the genus Chroomonas have been examined by electron microscopy. They demonstrate certain characteristic features of the Cryptophyceae, such as the presence of trichocysts and the arrangement of thylakoids in pairs in the plastid. The prominent pyrenoid of the genus Chroomonas is penetrated longitudinally by a tongue of cytoplasmic matrix which originates from between the two pairs of plastid membranes. In the genus Hemiselmis, however, the pyrenoid is traversed by a pair of thylakoids. The nucleus, Golgi apparatus and Corps de Maupas also occupy characteristic positions. These features indicate a close relationship between these two genera and the other major genus Cryptomonas, and support the suggestion that the Cryptophyceae is a discrete taxonomic group.     

Heterocapsa circularisquama sp. nov. (Peridiniales, Dinophyceae): A new marine dinoflagellate causing mass mortality of bivalves in Japan

Heterocapsa circularisquama Horiguchi sp. nov. is described from Ago Bay, central Japan. The dinoflagellate produced large-scale red tides in the bays of central and western Japan and caused mass mortality of bivalves, notably the pearl oysters. The cell is small and is composed of a conical epitheca and a hemi-spheroidal hypothecs. The chloroplast is single and is connected to the single pyrenoid. The nucleus is elongated and is located in the left side of the cell. Thecal plate arrangement has been determined as: Po, cp, 5′, 3a, 7″, 6c, 5s, 5″′, 2″″. Heterocapsa circularisquama is morphologically very similar to Heterocapsa illdefina and it is almost impossible to distinguish these two species at light microscopical level. The characteristics which can be used to distinguish these two species are the morphology of body scales and the ultrastructure of the pyrenoid matrix. The body scales of H. circularisquama possess six radiating ridges on the circular basal plate; no such ridges can be observed on the roughly triangular basal plate of the scales of H. illdefina. Furthermore, the scales of the latter species possess substantially shorter spines compared to those of H. circularisquama. The pyrenoid matrix of H. circularisquama is hardly perforated by cytoplasmic tubules, while in H. tlldefina the pyrenoid matrix is always penetrated by many cytoplasmic tubules. Based on the arrangement of thecal plates, morphology of body scales, and ultra-structure of the pyrenoid, I am placing H. circularisquama sp nov. into the genus Heterocapsa.     

Growth and reproduction of Scytosiphon lomentaria (Fucophyceae) in relation to temperature in two populations from Denmark

Two strains of Scytosiphon lomentaria from Denmark have been grown under long-day conditions on a temperature-gradient plate. The material from north-west Jutland (strain No. 46) has a reduced prostrate system (a few-celled lump) whereas the other material from the Sound (strain No. S41) has a well-developed crust. This difference plays a significant role in growth and reproductive patterns. Strain No. 46 rapidly produces a great number of cylindrical macrothalli. Within a four-week experimental period, formation of swarmers from plurilocular sporangia on the macrothalli takes place and results in a maximum dry weight of 32.9 mg at 18.2°C.
Swarmers from the macrothalli of strain S41 always develop into knot-filamentous or crust-shaped thalli. The development of the macrothalli, therefore, takes a longer time than in strain 46 and the maximum dry weight is only 16.5 mg at 16°C even within an experimental period of six weeks. The reproductive pattern is more complex than in strain 46. The crusts produce unilocular sporangia at temperatures between 16°C and 23.8°C, and function, therefore, as the reproductive system during the summer as macrothalli disappear at temperatures above 18°C both in nature and in culture. Formation of plurilocular sporangia on macrothalli starts when they are from 6–9 weeks old at temperatures between 11°C and 18°C.
The temperature range for growth in the two strains also differs: Strain 46 grows from 7.3–24.4°C while strain S41 grows at least from 4–5.1°C to an upper limit of 27.1°C, but lower limit has not been determined precisely in the present experiment. These results indicate that temperature ecotypes exist, even within a limited geographic area.     

CHLOROPLAST DIVISION AND PYRENOID FORMATION IN CHLAMYDOMONAS REINHARDI1

The process of chloroplast division during mitosis in Chlamydomonas reinhardi is followed with the electron microscope. The pyrenoid and the chloroplast reproduce by fission. The dividing chloroplast contains regions of dense material that superficially resemble pyrenoids, but it is concluded that the formation of the dense material is not related to pyrenoid formation in C. reinhardi. The dense material appears to be localized over regions of chloroplast DNA.     

STRUCTURE AND FUNCTION OF THE ALGAL PYRENOID. I. ULTRASTRUCTURE AND CYTOCHEMISTRY DURING ZOOSPOROGENESIS OF TETRACYSTIS EXCENTRICA1

The fine structure of the pyrenoid in the mature vegetative cell of Tetracystis excentrica Brown and Bold is described. During zoosporogenesis, the pyrenoid undergoes regression, and the ultrastructure of this process is described in detail. The ground substance undergoes dissolution, and reticulate fibrillar structures appear as well as intruding chloroplast thylakoids. Pyrenoid-associated starch plates diminish, and quantities of starch not associated with the pyrenoid are produced. New pyrenoids appear late in the division cycle after all other major organelles associated with the motile cell have been formed. Zoospore pyrenoids develop in thylakoid-free spaces of the chloroplast which are similar to the DNA-containing regions. The new pyrenoid ground substance, which is loosely fibrillar, arises in close proximity to starch grains which may be formed in the stroma. Then the zoospore pyrenoid produces 2 hemispherical starch plates identical to those in the mature vegetative cell. Zoospore pyrenoids lack the 2 convoluted thylakoids between the starch plates and the ground substance characteristic of those in the mature vegetative cell. Instead, the thylakoids are identical to those of the chloroplast at first, and then develop into a convoluted state in the vegetative cell. Cytochemical tests for DNA, RNA, and protein were made for the cytoplasm, nucleus, nucleolus, and pyrenoid. Conclusive evidence is presented for the presence of RNA in the cytoplasm and nucleolus, DNA in the nucleus, and protein in the pyrenoid. The tests did not conclusively demonstrate the presence or absence of DNA and RNA in the pyrenoid; however, they suggested that small amounts of both DNA and RNA may be present.     

The light organ and ink sac of Heteroteuthis dispar (Mollusca: Cephalopoda)

There has previously been some dispute over the source of the light of Heteroteuthis. Light and electron microscopy have shown that there are no bacteria in the cavity of the luminous gland, but rather a population of spherical vesicles of varying size and content, surrounded by dense interstitial material. The contents are secreted by the cells surrounding the gland via a microvillous brush border that contains cilia. The gland is surrounded by a reflector layer made up of iridophores, whose structure differs considerably in the proximal, lateral and cap regions.
The ink sac contains masses of spherical dense granules and its wall cells have many microvilli extending amongst these granules.