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.     

AXILOSPHAERA AND HETEROTETRACYSTIS,NEW CHLOROSPHAERACEAN GENERA FROM TENNESSEE SOIL12

Two new chlorosphaeracean genera were isolated into axenic culture from soil collected in cedar glades in Cedars of Lebanon State Forest, Wilson County, Tennessee. The distinguishing characteristics of the new monotypic genus Axilosphaera include an axile (asymmetric) ckloroplast with at least 1 pyrenoid and Chlamydomonas-type (walled) zoospores. A. vegetata is the type species. Reproduction is by dissociation of daughter cells following vegetative cell division, by zoospores, and by aplanospores. The new polytypic genus Heterotetracystis, comprising 3 species, H. akinetos, H. macrogranulosa, and H. intermedia, is characterized by a parietal chloroplast with at least 1 pyrenoid and walled zoospores with flagella of unequal length. Reproduction is by dissociation of daughter cells following vegetative cell division and by zoospores. H. akinetos is designated as the type species.     

HEMIFLAGELLOCHLORIS KAZAKHSTANICA GEN. ET SP. NOV.: A NEW COCCOID GREEN ALGA WITH FLAGELLA OF CONSIDERABLY UNEQUAL LENGTHS FROM A SALINE IRRIGATION LAND IN KAZAKHSTAN (CHLOROPHYCEAE,CHLOROPHYTA)1

A coccoid green alga, Hemiflagellochloris kazakhstanica S. Watanabe, S. Tsujimura, T. Misono, S. Nakamura et H. Inoue, gen. et sp. nov., was described from soil samples of a saline irrigation land in Ili River basin, Kazakhstan. This alga had a parietal chloroplast with a pyrenoid, which was covered with starch segments and penetrated with thylakoid membranes. Reproduction occurred by the formation of aplanospores and zoospores. The aplanospores frequently formed tetrad aggregations in a mother cell. The zoospores were covered by a single‐layered cell wall and lacked stigmata. The zoospores had two flagella of considerably unequal lengths; the longer flagellum was 17–19 lm in length and the shorter one was 9–10 lm. The flagellar apparatus architecture was of the clockwise orientation group type in the Chlorophyceae. Molecular phylogenetic analysis using 18S and 28S rDNA sequence data resolved this organism in a separate clade from the green algae that had flagella of slightly unequal lengths. It was suggested that features such as inequality in flagellar lengths, parallel exsertion of basal bodies, and subapical position of the flagellar apparatus were sporadically evolved.     

MORPHOLOGY AND PAEDOGAMOUS SEXUAL REPRODUCTION IN CHLOROGONIUM CAPILLATUM SP. NOV. (VOLVOCALES, CHLOROPHYTA)1

Morphology and sexual reproduction in Chlorogonium capillatum Nozaki, Watanabe & Aizawa sp. nov. (Volvocales, Chlorophyta) originating from Miyatoko Mire, Japan, were studied under controlled laboratory conditions. Vegetative cells of this new species were fusiform with blunt anterior and posterior ends, and they had a massive parietal chloroplast and numerous contractile vacuoles distributed throughout the protoplast. Several to many pyrenoids were randomly distributed in the chloroplast, but they disappeared under the light microscope when grown photoheterotrophically. During asexual reproduction, the first division took place transversely without a preceding rotation of the parental protoplast. In sexual reproduction, the parental protoplast divided successively to form 32 or 64 small, biflagellate isogametes. After gametogenesis, the gametes did not escape from the parental cell (gametangial) wall, within which pairs of the adjoining gametes fused to form quadriflagellate zygotes. Such zygotes were then released from the parental cell wall and developed into hypnozygotes, which at maturity developed numerous thin spines or hairs on the zygote wall. On zygote germination, four biflagellate germ cells were released from the zygote wall separately. This type of gametic union, "paedogamy," has not previously been described in the green algae except for Chlorococcum echinozygotum Starr . Chlorogonium capillatum can be clearly distinguished from other described species of Chlorogonium by its numerous contractile vacuoles and blunt anterior and posterior ends in vegetative cells as well as by its unique sexual reproduction, in which paedogamous conjugation occurs, and numerous thin spines or hairs that develop on the hypnozygote walls .     

GLOEOCOCCUS TETIMSPORUS SP. NOV. (TETRASPORALES,PALM ELL ACEAE), FROM COLORADO MOUNTAIN LAKES1

A new species, Gloeococcus tetrasporus sp. nov., collected from mountain lakes, is described from unialgal culture. Vegetative cells are ellipsoid and Chlamydomo nas–like, occur in tetrad complexes within the general colonial matrix, and exhibit slow, limited motility within the confines of the individual gelatinous matrices. The colonial matrix is amorphous and structureless, without a definite bounding layer. Colonies may reach several centimeters in size. Vegetative cells have a parietal cup–shaped chloroplast with a central–basal pyrenoid and a small, linear stigma, two contractile vacuoles, and two short flagella. Cell division is by eleutheroschisis in nonflagellate cells. After two divisions, four daughter cells arc formed within the expanded parent wall that will become incorporated into the colonial matrix. Zoospores are formed either from transformed vegetative cells or after cytokinesis. Zoospore flagella are two to three times the length of vegetative cell flagella. Rapid flagellar movement ruptures the sheath and liberates the zoospores. When zoospores settle, they secrete new sheaths, and divide twice to initiate new colonies. Sexual reproduction and formation of resistant spores were not observed.     

真眼点藻纲的系统分类、生物学特性及应用研究

真眼点藻纲是1970年、1971年Hibberd 和 Leedale根据其细胞学和超微结构的特征, 将原黄藻纲中的一些成员重新调整而成立的一个新纲。自该纲成立以来, 由于新种不断被发现, 科、属和种的数量都有所增加, 目前该纲已有1目6科13个属28个种。该纲藻类的主要细胞光合作用色素包括: 叶绿素a、堇菜黄素、无隔藻黄素和-胡萝卜素; 细胞内具有一周生裂叶状的叶绿体或多个盘状的叶绿体, 具一柄状蛋白核或无, 叶绿体内具有三条类囊体为一组的片层, 无环带形的片层, 双层叶绿体膜外有一层叶绿体内质网膜包裹, 它不与核膜相连; 细胞中具有一个相对较大的、近球形的液泡, 其中含有能振动的颗粒物和一个直径在1-3 m的红色球状体; 繁殖方式通常形成2个D形或4个四面体形的似亲孢子, 有时会形成8个或16个的似亲孢子; 大多种类能产生烧瓶状的游动孢子, 游动孢子具有单根两侧排列管状小茸毛的鞭毛（另一根退化）或另具一根为光滑型的鞭毛。通过18S rDNA和rbcL基因序列的分析, 该纲与异鞭藻门其他各纲藻类的亲缘关系得到确定。脂肪酸分析结果发现该纲的藻类皆含有长链多不饱和脂肪酸-二十碳五烯酸。真眼点藻纲的藻类在淡水、海水和土壤表面等环境中都有分布。该纲的拟微绿球藻属中多个种类被广泛应用于轮虫、卤虫及珍贵海产品幼苗的开口饵料, 亦已成为二十碳五烯酸和重要类胡萝卜素的潜在生产藻株。另外, 研究发现该纲的大多数种类富含油脂, 它们已成为微藻生物燃料开发的备选藻种资源。       

THE RELEASE,SETTLEMENT AND GERMINATION OF ZOOSPORES IN CHORDA TOMENTOSA (PHAEOPHYCEAE,LAMINARIALES)1,2

Details of zoospore germination in Chorda tomentosa Lyngb. are outlined. Uninucleate zoospores, when released are embedded in a mucilaginous mass of carbohydrate which dissolves and the biflagellate zoospores become motile. The long anterior flagellum is composed of a highly coiled terminal region and a rigid lower section bearing mastigonemes. The rigid, short posterior flagellum lacks mastigonemes. After initial contact by the tightly coiled region of the anterior flagellum, the zoospore draws itself to the substrate by flagellar resorbtion. After deposition of 3 wall layers the germling produces a germ tube. During this time the disc-shaped chloroplast enlarges undergoing changes in shape. As the germ tubes reach ca. 15 μm they cease forward growth and swell at their tips. The majority of cytoplasm of the original zoospore moves into the tube. Just before the nucleus enters the tube, centriole replication occurs. Mitosis is presumed to take place somewhere in the germ tube so that at 24 h, 2-celled gametophytes are produced.     

<Emphasis Type="Italic">Norrisiella sphaerica</Emphasis> gen. et sp. nov., a new coccoid chlorarachniophyte from Baja California,Mexico

A new chlorarachniophyte, Norrisiella sphaerica S. Ota et K. Ishida gen. et sp. nov., from the coast of Baja California, Mexico is described. We examined its morphology, ultrastructure, and life cycle in detail, using light microscopy, transmission electron microscopy, and time-lapse videomicroscopy. We found that this chlorarachniophyte possessed the following characteristics: (1) vegetative cells were coccoid and possessed a cell wall, (2) a pyrenoid was slightly invaded by plate-like periplastidial compartment from the tip of the pyrenoid, (3) a nucleomorph was located near the pyrenoid base in the periplastidial compartment, (4) cells reproduced vegetatively via autospores, and (5) a flagellate stage was present in the life cycle. This combination of characteristics differs from any of the described chlorarachniophyte genera, and therefore a new genus is established. Fluorescent microscopic observations suggested that the alga formed multinucleate cells prior to forming autospores. Time-lapse observations during autospore formation showed that cytokinesis occurred simultaneously in the multinucleate cells. Zoospores were also produced, and video sequences captured the release of zoospores from coccoid cells. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

FINE STRUCTURE OF ZOOSPOROGENESIS, ZOOSPORE GERMINATION, AND EARLY GAMETOPHYTE DEVELOPMENT IN CLADOPHORA SURERA (CLADOPHORALES, CHLOROPHYTA)

The fine structure of zoosporogenesis, zoospore germination, and early gametophyte development in Cladophora surera Parodi et Cáceres were studied. Zoosporogenesis started with simultaneous meiosis in all nuclei of apical initial cells. The resulting haploid nuclei duplicated in turn by successive centric, closed mitoses. Then, each initial cell divided into two short zoosporangia. Numerous vacuoles appeared around each sporic nucleus. The delimitation of uninucleate zoosporocytes occurred by cytokinetic furrows produced by the coalescence of tiny, clear vesicles, without microtubules. Final shape of the zoospore resulted from gradual expulsion of vacuoles from the cell body. Mature biflagellate zoospores exhibited a conspicuous apical papilla containing fine granular globules, the basal apparatus, and a microtubular "umbrella" formed by numerous cortical microtubules that ran backward the length of the cell body. The chloroplast showed a conspicuous eyespot. The zoosporangial wall disorganized at the pore through which the zoospores were liberated. Zoospores settled on a substrate by their anterior papilla secreting an adhesive. Germination involved retraction of the apical papilla, loss of the "umbrella" microtubules and eyespot, and the lateral absorption of the entire flagellar apparatus, i.e. basal apparatus plus axoneme, into the cytoplasm. Early gametophyte development involved the synthesis of a thin, young cell wall, the development of outer peripheral vacuoles, the appearance of the marginal reticulate chloroplast, and the formation of the first central vacuoles derived from abundant endoplasmic reticulum. Close to the plasmalemma ran longitudinally oriented cortical microtubules. Eventually, the germling developed an achlorophylic, elongated rhizoidal portion.     

ATRACTOMORPHA ECHINATA GEN. ET SP. NOV., A NEW ANISOGAMOUS MEMBER OF THE SPHAEROPLEACEAE (CHLOROPHYCEAE)1,2

Atractomorpha echinata gen. et sp. nov. is described from isolates derived from zygotes present in a dry soil sample obtained from Texas. The new genus is distinguished from Sphaeroplea primarily by its pattern of vegetative growth. While Sphaeroplea is distinctly filamentous with numerous coenocytic cells uniseriately arranged, Atractomorpha grows as individual, multinucleate, spindle-shaped cells with sharply pointed extremities. Such cells may vary considerably in length (25–6000 μm, or more) and normally lack septa. In young, rapidly growing cultures the cells often attain lengths of 300–500 μm, but rarely exceed 1800 μm. The new species is further characterized by: (1) the regular formation of biflagellate zoospores in asexual reproduction, (2)anisogamy (occasionally oogamy) and (3) the size and ornamentation of its zygotes. Variations in vegetative morphology are discussed as are conditions for obtaining gametogenesis.     

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.     

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.     

Nematophagous fungi: three new species of Myzocytium.

Three species of Myzocytium parasitic on nematodes are described as new. In M. papillatum the zoospores encyst directly on the host cuticle before penetration. This species produces smooth, spherical oospores. In M. glutinosporum the biflagellate zoospores do not attack the host directly; after encystment they produce a spherical adhesive bud which allows the spores to adhere to the cuticle of passing nematodes. This species produces echinulate, spherical oospores. In M. anomalum the primary spores are aplanospores. After a dormant phase, and when suitably stimulated, these aplanospores change into biflagellate zoospores and the latter encyst on the host cuticle. No sexual state is known in this species. Persistence is by means of thick-walled, spherical chlamydospores.     

Studies on the vegetative life cycle of Chlamydomonas reinhardi Dangeard in synchronous culture I. Some characteristics of the cell cycle

Cells of Chlamydomonas reinhardi Dangeard were grown synchronouslyunder a 12 hr light-12 hr dark regime. Time courses of nucleardivision, chloroplast division, "apparent cytokinesis" and zoosporeliberation were followed during the vegetative cell cycle inthe synchronous culture. Liberation of zoospores occurred atabout 23–24 hr after the beginning of the light periodat 25°C. Four zoospores were produced per mother cell underthe conditions used. At lower temperatures, the process of zoosporeliberation as well as length of the cell cycle was markedlyprolonged, but the number of zoospores produced per mother cellwas approximately the same. At different light intensities,lengths of the cell cycle were virtually the same, while thenumber of zoospores liberated was larger at higher rather thanat lower light intensities. During the dark period, nuclear division, chloroplast divisionand apparent cytokinesis took place, in diis order, and proceededless synchronously than did the process of zoospore liberation.When the 12 hr dark period was replaced with a 12 hr light periodduring one cycle, the time of initiation as well as the durationof zoospore liberation was litde affected in most cases, whereasnuclear division, chloroplast division and apparent cytokinesiswere considerably accelerated by extended illumination. Whenalgal cells which had been exposed to light for 24 hr were furtherincubated in the light, zoospore liberation started much earlierand proceeded far less synchronously, compared with that under12 hr light-12 hr dark alternation. (Received October 12, 1970; )     

TWO TYPES OF CYTOPLASMIC CLEAVAGE IN THE COENOCYTIC SOIL ALGA PROTOSIPHON BOTRYOIDES (CHLOROPHYCEAE)1

Cytokinesis in the coenocytic green alga Protosiphon botryoides (Kütz.) Klebs was studied with transmission electron microscopy. In vegetative cells, nuclei with associated basal bodies and dictyosomes are scattered throughout the cytoplasm. Mature cells may develop either multinucleate resting spores (coenocysts) or uninucleate zoospores. Cytokinesis may be preceded by contraction of the protoplast due to the disintegration of vacuoles that are present in larger, siphonous cells. The formation of coenocysts in ageing, siphonous cells, is signalled by cleavage of the chloroplast and the development of arrays of phycoplast microtubules in one or more transversely oriented planes through the cell. Nuclei with associated basal apparatuses stay dispersed throughout the cytoplasm; the basal bodies apparently are not involved in organization of the phycoplast. The plasma membrane invaginates, resulting in a centripetal cleavage of the protoplast into two or more multinucleate daughter protoplasts. Simultaneously, wall material is deposited along the outside of the daughter protoplasts by dictyosome-derived vesicles, and finally two or more thick-walled coenocysts are formed. The formation of zoospores, on the other hand, is signalled by clustering of the nuclei in one or more groups depending on the shape of the mother cell. The nuclei become arranged with the associated basal apparatuses facing toward the center of the cluster. Bundles of phycoplast microtubules develop between the nuclei, radiating from the center of a cluster toward the plasma membrane; basal apparatuses or associated structures apparently are involved in organization of the phycoplast. Cleavage furrows grow out centrifugally along these bundles of micro-tubules, fed by dictyosome-derived vesicles. No wall material is deposited. An additional mitotic division occurs during cleavage, and finally numerous uninucleate, wall-less, biflagellate zoospores are formed. The ultrastructural features of the two different types of cytoplasmic cleavage associated with two different types of daughter cells have not previously been reported for chlorophycean algae.     

1株裂殖壶菌(Schizochytrium sp.)的分离鉴定

采用松树花粉诱集法从乐清湾红树林分离到一株纯培养物,其特点为：营养菌体为椭圆球形,单核;营养菌体的细胞壁由许多紧压在一起的致密鳞片层构成,在细胞壁不连续处可分辨鳞片;营养菌体形成外质网,它产生于外质网形成体;营养菌体以产生游动孢子行无性繁殖,游动孢子为双鞭毛;无性繁殖过程中形成四分体结构。据此鉴定为裂殖壶菌（Schizochytrium sp．）。     

PSEUDOTETRACYSTIS,A NEW CHLOROSARCINACEAN ALGA12

Pseudotetracystis terrestris gen. et sp. nov., a member of the Chlorosarcinaceae, was isolated into axenic culture from dry surface soil collected near Texon in Reagan County, Texas. The characteristics of this presently monotypic genus include vegetative cells with a parietal chloroplast, a single pyrenoid, and polymorphic motile cells which are of the Protosiphon-type (naked with 2 equal flagella); P. terrestris is the type species. Vegetative cell division (desmoschisis) forms diads and tetrahedral or planar tetrads, sometimes aggregated in transitory complexes. Reproduction is by the polymorphic motile cells which may function either as gametes to form zygotes which develop without dormancy directly into vegetative cells, or they may function as zoospores and develop directly into vegetative cells without union.     

OBSERVATIONS ON THE FINE STRUCTURE OF OEDOGONIUM. V. EVIDENCE FOR THE DE NOVO FORMATION OF PYRENOIDS IN ZOOSPORES OF OE. CARDIACUM123

Ultrastructural studies of the chloroplasts of zoospores and developing zoospores of Oedogonium carcliacum have disclosed the occurrence of numerous incipient pyrenoids. A single developing zoospore may possess several score of these structures which appear to arise de novo in the chloroplast stroma and seem to lack any direct association with mature pyrenoids which are also present in the cells. The incipient pyrenoids lack the associated starch grains and the membrane-limited channels characteristic of mature pyrenoids, but they are readily recognized in the chloroplasts since they demonstrate a greater granularity and electron density than the surrounding chloroplast stroma. The granularity and electron density of the incipient pyrenoids match the ultra-structural appearance of the matrix of mature pyrenoids. The smallest of the incipient pyrenoids examined from serial sections had a maximum diameter of less than 0.3 μ. This may be compared with the size of mature pyrenoids, many with a maximum diameter of over 5.0 μ. In all the zoospores and developing zoospores examined, only one mature pyrenoid was observed in an apparent stage of division.     

The effect of colchicine on colony formation in the algae Hydrodictyon,Pediastrum and Sorastrum

Summary Uninucleate, biflagellate zoospores of Hydrodictyon, Pediastrum and Sorastrum, derived from multinucleate parental cells, aggregate and adhere to form distinctively patterned colonies; earlier work has shown that microtubules underlie the plasmalemma of these zoospores and are also conspicuous in the developing horns of aggregating cells of Pediastrum and Sorastrum. Colchicine applied to parental cells inhibited cytoplasmic cleavage and production of uninucleate zoospores. When zoospores were treated with colchicine, their peripheral microtubules disappeared; the spores failed to aggregate in ordered arrays and did not develop horns. The microtubules therefore appear to play an important role in determining the arrangement of cells in developing colonies by affecting the shape of the cells at the time of their aggregation.