LIGHT AND ELECTRON MICROSCOPIC CHARACTERIZATION OF TWO TYPES OF PYRENOIDS IN GONIUM (GONIACEAE,CHLOROPHYTA)1

The single, basal pyrenoids of Gonium quadratum Pringsheim ex Nozaki and G. pectorale Müller (Goniaceae, Chlorophyta) differed in appearance when vegetative colonies were cultured photoheterotrophically in medium containing sodium acetate. Chloroplasts of G. quadratum had distinct pyrenoids when grown in medium without major carbon compounds. However, the pyrenoids degenerated and were markedly reduced in size when such cells were inoculated into a medium containing 400 mg·L^?1 of sodium acetate. No pyrenoids were visible under the light microscope; however, with electron microscopy small pyrenoids and electron-dense bodies were visible within the degenerating chloroplasts, which had only single layers of thylakoid lamellae at the periphery. The chloroplasts subsequently developed distinct pyrenoids and several layers of thylakoid lamellae as the culture aged. In contrast, vegetative cells of G. pectorale always showed distinct pyrenoids when cells were inoculated into medium containing sodium acetate, sodium pyruvic acid, sodium lactate, and/or yeast extract. Therefore, we propose two terms, “unstable pyrenoids” and “stable pyrenoids,” for pyrenoids of G. quadratum and G. pectorale, respectively. Chloroplasts of the colonial green flagellates should thus be examined under various culture conditions in order to determine whether their pyrenoids are unstable or stable when pyrenoids are used as taxonomic indicators. Immunogold electron microscopy showed that the ratios of gold particle density of ribulose-1,5-biphosphate carboxylase/oxygenase (RuBisCO) between pyrenoid matrix and chloroplast stroma in G. quadratum grown in medium with or without sodium acetate were lower than those of G. pectorale. Heavy labeling by anti-RuBisCO was observed in both the electron-dense bodies and pyrenoid matrix of G. quadratum. This is the first electron microscopic demonstration of degeneration and development of both pyrenoids and thylakoid lamellae in the chloroplast as a function of culture condition in green algae.     

泡状饶氏藻营养细胞的超微结构研究

本文报道我国特有的一种绿藻门植物－泡状饶氏藻营养细胞的超微结构特征,植物体由3层细胞组成,外层细胞最小,细胞质比较丰富,含有较多的各种细胞器,液泡体积较小,中层细胞具有很大的中央液泡,细胞质成为贴壁的薄层,各种细胞器结构仍清晰可见,内层细胞极度液泡化,细胞质呈现退化状态,周生的片状叶绿体上有许多大小不等的穿孔,使叶绿体呈网孔状外貌,叶绿体主要由许多成对的类囊体组成,叶绿体上往往有几个蛋白核,蛋白核经常被1或2条类囊体穿过,呈现分隔状,本文也报道了泡状饶氏藻的线粒体,质体,内质网,高尔基体和核内微管的结构特征,根据泡状饶氏藻的类囊体形态与Ulva mutabilis非常相似以及蛋白核的超微结构特征,它与石莼科植物可能有较密切的亲缘关系,属于绿藻门中进化的类群。     

Ultrastructural Studies of Vegetative Cells on Jaoa bullata (Jao) Fan

The present study reports the ultrastructural features of the vegetative cells in Jaoa bullata (Jao), Fan., an endemic species of Chlorophyta in China. Its thallus is composed of 3 layers of cells. Outer cells are the smallest in size, containing abundant cytoplasm, rich in various organelles; their vacuoles are smaller. The cells of mid-layer possess a large central vacuole. Their cytoplasm becomes a thin layer appressed against the cell wall; Various kinds of the organelles are still clearly visible. Inner ceils are extremely vacuolated. Their cytoplasm looks like degenerated. The cells are uninucleate, each containing a parietal, laminate chloroplast with numerous pores, which cause chloroplast a net-like appearance. The photosynthetic lamellae in a chloroplast include a number of thylakoids mainly in pairs. Chloroplast contains several pyrenoids, which are penetrated by 1 or 2 thylakoids. The present study deals with the structural characteristics of the mitochondria, plastids, endoplasmic reticulum and dictyosomes in the cells of Jaoa bullata. Two groups of intranuclear microtubules are present in a cell. Based on the similarity to Ulva mutabilis in the thylakoid arrangement and the ultrastructural features of the pyrenoids, authors suggest that daoa bullata (Jao) Fan may be closely related to Ulvaceae, among the advanced taxa in the evolution of Chlorophyta.     

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.     

CYTOLOGY AND ULTRASTRUCTURE OF CHLORARACHNION REPTANS (CHLORARACHNIOPHYTA DIVISIO NOVA,CHLORARACHNIOPHYCEAE CLASSIS NOVA)1

The green amoeboid cells of Chlorarachnion reptans Geitler are completely naked and each contains a central nucleus, several bilobed chloroplasts each with a central projecting pyrenoid enveloped by a capping vesicle, several Golgi bodies, mitochondria with tubular cristae, extensive rough ER, and a distinct layer of peripheral vesicles. Complex extrusome-like organelles occur rarely in both the amoeboid and flagellate stages. The only organelles entering the reticulopodia are mitochondria, but microtubules are also present. The chloroplasts contain chlorophylls a and b, but histochemical tests suggest that the carbohydrate storage product probably is not a starch. The chloroplast lamellae are composed of one to three thylakoids or form deep stacks. A girdle lamella and interlamellar partitions are absent. Each chloroplast is bounded by either four separate membranes, a pair of membranes with vesicular profiles between them, or three membranes; all three arrangements may occur in the same chloroplast. A periplastidal compartment occurs near the base of the pyrenoid where there are always four surrounding membranes. The compartment has a relatively dense matrix and contains ribosome-like particles and small dense spheres; it extends over and into a deep invagination in the pyrenoid where its contents are enclosed in a double-membraned envelope which is penetrated by wide pores. The zoospores are ovoid and each bears a single laterally inserted flagellum which appears to be wrapped helically around the cell body during swimming. The flagellum lies in a groove in the cell surface and bears fine lateral hairs. Neither a second flagellum or vestige of one, nor an eyespot, is present. A single microtubular root and a larger homogeneous root run from the flagellar base parallel to the emerging flagellum, between the nuclear envelope and the plasmalemma. In the simple flagellar transition region, fine filaments connect adjacent axonemal doublets. A detailed comparison of C. reptans with all other algal taxa results in the conclusion that it must be segregated in the new class Chlorarachniophyceae, the only class in the new division Chlorarachniophyta. The possibility that C. reptans evolved from a symbiosis between a colorless amoeboid cell and a chlorophyll b- containing eukaryote is considered, but the possible affinities of the symbiont remain enigmatic. The implications of the unique chloroplast structure of C. reptans for current hypotheses concerning the origin of chloroplasts are discussed.     

几种藻类蛋白核的超微结构研究

应用电镜及免疫电镜技术对莱茵藻、小球藻、条浒苔和紫菜等藻类的叶绿体蛋白核的超微结构及主要组成成分进行了观察和研究。结果显示：不同藻类的蛋白核结构不同,显示了藻类蛋白核的多样性。蛋白核为球形或椭圆形,由蛋白质组成。莱茵藻、小球藻和条浒苔的蛋白核外围被淀粉鞘所包围,而紫菜的蛋白核外围无淀粉鞘而直接被叶绿体的类囊体所包围。淀粉鞘由淀粉组成,淀粉鞘的厚薄与藻体藻龄及增养状态有关系。在蛋白核中央,一般都具有由类囊体形成的孔道,使蛋白核与外界联系,小球藻和条浒苔蛋白核具有1条纵向孔道,而莱茵藻和紫菜为多条孔道。金相免疫技术检测结果显示Rubisco和Rubisco活化酶均在蛋白核及淀粉鞘区域中定位,表明蛋白核具有光合作用功能．     

THE ULTRASTRUCTURE OF THE VITELLINE BODY IN THE OOCYTE OF THE SPIDER TEGENARIA PARIETINA

The vitelline body in the mature oocyte of the spider Tegenaria parietina is composed of 4 different zones. 1. The central zone contains granular areas, vesicles, and a few lamellae. 2. The lamellar zone consists of numerous concentric lamellae. These sheets, 45 A in thickness, are stacked in groups. The fine structure and the regular arrangement recall those of myelin sheets, retinal rods, and chloroplasts. Between the stacks of lamellae, finely granular masses and various vesicles are to be found. 3. The "zone of transition" consists of a finely granular substance accumulated in abundant masses. This substance is composed of very closely packed granules about 50 to 60 A in diameter. Very often, near the lamellae, the granules show alignment giving a gradual transition from grains to lamellae. 4. The vesicular zone contains ergastoplasm, dense particles, mitochondria, and Golgi material. It is suggested that the peculiar ultrastructure of these cytoplasmic components may be related to an intense metabolic activity.     

EFFECTS OF DARKNESS AND OF STREPTOMYCIN ON THE FINE STRUCTURE OF EUGLENA GRACILIS

Siegesmund , Kenneth A., Walter G. Rosen , and Stanley R. Gawlik . (Marquette (J., Milwaukee, Wis.) Effects of darkness and of streptomycin on the fine structure of Euglena gracilis. Amer. Jour. Bot. 49 (2) : 137–145. Illus. 1962.—Dark-grown Euglena gracilis cells, transferred from streptomycin (SM)-containing medium to SM-free medium 5 days before transfer to light, turn green normally, indicating that proplastids are unaffected by SM. SM-bleached cells, grown in light, contain numerous bodies composed of concentric lamellae (CL bodies). These differ from chloroplasts in that their lamellae lack electron-dense dots, are not coalesced, and they lack a 3-layered limiting membrane and pyrenoids. CL bodies are absent from dark-grown normal and dark-grown SM-bleached cells, as well as from light-grown normal cells. It is suggested that CL bodies result from a derangement of chloroplast synthesis caused by SM blockage of chlorophyll synthesis.     

OBSERVATIONS ON THE FINE STRUCTURE OF DICTYOCHA FIBULA EHRENBERG. II. THE PROTOPLAST1

Dictyocha fibula in exponential phase cultures displays a range of morphological variants of which the “sunburst” is most common. In this form, the perinuclear cytoplasm (perikaryon) contains an average of 72 dictyosomes, assorted vesicles, mitochondria, endoplasmic reticulum, and ribosomes. Cytoplasmic processes, globose to irregular, extend on fine cytoplasmic strands from the perikaryon into an extensive, viscous wall, structureless in electron micrographs except for scattered electron-opaque leaflets near the perikaryon. Mitochondria with tubular cristae occur within the globose process and occasionally within the connecting strands. Chloroplasts, with 3-disk lamellar bands and with pyrenoids not crossed by lamellae, are confined to the cytoplasmic processes in the sunburst from. A structure which may be the “flagellum” occasionally occurs attached to the perikaryon. However, no flagellar structures containing microtubules, nor flagellar root structures, have been found.     

ELECTRON MICROSCOPY OF THE VOLVOCACEAE AND ASTREPHOMENACEAE

Lang, Norma J. (U. Texas, Austin.) Electron microscopy of the Volvocaceae and Astrephomenaceae. Amer. Jour. Bot. 50(3): 280-300. Illus. 1963.—Clonal cultures of Gonium sociale, G. pectorale, Pandorina morum, Eudorina elegans, Eudorina sp., Volvulina steinii, V. pringsheimii, Platydorina caudata, Pleodorina illinoisensis, P. californica, Volvox aureus, V. tertius, V. globator, V. barberi, and Astrephomene gubernaculifera representing the Volvocaceae and Astrephomenaceae in the Volvocales were examined with the electron microscope and their ultrastructure compared. The ultrastructure of the various organelles is basically similar in the species studied and no increase in cellular complexity is found to accompany the evolutionary trends evidenced in the Volvocaceae. The ultrastructure of a colonial cell is basically that of Chlamydotnonas. A cytoplasmic membrane having a unit membrane structure encompasses a cell and is continuous with the double-membraned flagellar sheaths. The flagella contain the typical 9 + 2 fibril arrangement with the 2 axial fibrils terminating in a cylinder at the flagellar base and the 9 peripheral pairs continuing into the cytoplasm as a basal body. The organelles comprising the cytoplasm are: mitochondria with plate-like cristae; dictyosomes composed of stacks of agranular cisternae; small, rough or smooth-surfaced vesicles; an endoplasmic reticulum of granule-bearing and agranular tubules, lamellae and broad cisternae; vacuoles which are either contractile, contain fine granular and fibrillar material, or have dense contents probably representing polyphosphate; lipid bodies; and dense granules 100–150 A which have been called ribosomes. The finely granular nucleoplasm is surrounded by a porous, double-membraned nuclear envelope and contains a centric nucleolus composed of dense, spherical granules. The outer membrane of the nuclear envelope bears granules and may have granular extensions into the perinuclear cytoplasm. Each extension appears to encompass one or several dictyosomes and has been termed an “amplexus.” The amplexi are agranular on the surface contiguous to a dictyosome. A double-membraned chloroplast envelope is continuous around the single, cup-shaped chloroplast. The basic chloroplast units are discs closed at each end, occurring in stacks of varying number parallel to the envelope. The presumed proteinaceous matrix of the basal pyrenoid is penetrated by elongated, tubular elements which connect with the lamellar discs. Multiple rows of granules, associated with individual discs, form the anterior stigma within the chloroplast envelope. The colonial matrix is not a structureless, mucilaginous material uniting the cells in colonies, but it has rather a highly complex structure especially around the periphery of the colony and the flagellar channels. The apparent substitution of a fibrillar layer of the colonial matrix for the discrete compact cell wall, such as is found in Chlamydomonas, implies a greater degree of complexity in the evolution of these colonial genera than is generally assumed.     

Plastid Development and Ultrastructure in Yellowish Mutants of Sugar Cane

The comparisons of the ultrastructures of plastids in yellowish mutant and yellowish-green striped mutant and in normal green plant from tissue culture of sugar cane were made. There was no difference found in the structure and development of chloroplasts between the normal green leaves and the green tissue of striped leaves, but the plastid in the yellowish tissue of two kinds of mutants were anomalous. They had not a fully developed system of grana and stroma lameilae as in the normal green leaves. This aberrant plastid contained only some vesicles, a few lamellae and more or less clearly defined ribosome particles and DNA-like mierofibrils, while some stacking and swelling of thylakoids were often observed. In some sections of this aberrant plastid a bunchy lamellae and cross connective fibrils between parallel lamellae were often found too. However, the mixed cell which contains both of normal chloroplast and defective plastid together was never found in the leaves of the mutant plants. It was suggested that yellowish and yellowish-green striped leaves from tissue culture of sugar cane might be caused by nuclear gene mutation.     

Fine Structure of Peridinium westii Lemm., a Freshwater Dinoflagellate

SYNOPSIS. The fine structure of Peridinium westii Lemm., an armoured dinoflagellate collected from Lake Tiberias, is reported. Thick cell walls contain plates, pores and tubules, and are surrounded by an external membrane. The cytoplasm is limited by 3 membranes. Nuclei contain numerous chromosomes with a fibrillar organization. The chloroplast envelope consists of 3 membranes; parallel lamellae, each generally comprising 3 thylakoids, are present. Pyrenoids are present in various forms within the chloroplast structure. Trichocysts are found in cells both as elongate dense cores within a membrane-enclosed sac and as striated rods. An eyespot is found in a plastid containing several thylakoids.     

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.     

LIGHT AND ELECTRON MICROSCOPY OF PYRENOIDS AND SPECIES DELIMITATION IN VOLVULINA (CHLOROPHYTA,VOLVOCACEAE)1

The appearances of pyrenoids in the vegetative cells of Volvulina steinii Playfair and V. pringsheimii Starr were observed in detail by light and electron microscopy in relation to the culture age to clarify the taxonomic relationship between the two species. In V. pringsheimii, the pyrenoids were always present in the bottom of the cupshaped chloroplasts and their gross morphology did not vary in relation to the culture age, while those of V. steinii appeared de novo and developed as the culture aged. In 24-h cultures of V. steinii, pyrenoids were not observed in the chloroplasts. In 48-h cultures, a pyrenoid matrix developed apparently de novo in the brim of the cupshaped chloroplast. Subsequently, starch grains appeared around the pyrenoid matrix in 72-h cultures. The volume of the matrix and the associated starch grains increased and tubular channels entered into the pyrenoid matrix in 96-h cultures. In addition, the pyrenoid in the parental chloroplast of V. pringsheimii divided and was distributed to each daughter cell during cell divisions in daughter colony formation, while the parental pyrenoid of V. steinii did not divide and went to one of the daughter cells. Therefore, these two species can be clearly distinguished by the differences in the position of pyrenoids in the cupshaped chloroplasts and stability of pyrenoid appearance in relation to the culture age, as well as in the fate of parental pyrenoids during daughter colony formation.     

A DESCRIPTIVE AND ULTRASTRUCTURAL STUDY OF THE GREEN ALGA,CHLAMYDOMONAS DESMIDII SP. NOV. (VOLVOCALES. CHLAMYDOMONADACEAE) FROM ROCKY MOUNTAIN NATIONAL PARK LAKES 1

The green algal flagellate Chlamydomonas desmidii sp. nov., is described from several lakes in Rocky Mountain National Park, Colorado. The biflagellate cells generally have an hour-glass shape. They possess a single chloroplast with two pyrenoids and a superficial stigma near the center of the cell. Numerous contractile vacuoles occur in the Peripheral cytoplasm and they are arranged in several rings around the nucleus in the constricted region of the cell. Ultrastructural studies verify these features and elaborate upon the structure of the cell.     

TOWARD A NATURAL SYSTEM OF SPECIES IN CHLOROGONIUM (VOLVOCALES, CHLOROPHYTA): A COMBINED ANALYSIS OF MORPHOLOGICAL AND rbcL GENE SEQUENCE DATA

The taxonomy of species of Chlorogonium (Volvocales, Chlorophyta) was studied based on comparative light and electron microscopy and DNA sequence data of 23 strains from five major algal culture collections. All of the 23 strains showed pyrenoids under photoautotrophic conditions, but 17 of the 23 exhibited marked reduction in size of pyrenoids, or pyrenoids were absent under photoheterotrophic conditions. The strains could clearly be delineated into six species, C. euchlorum, C. elongatum, C. fusiforme, C. capillatum, C. neglectum, and C. kasakii on the basis of differences in cell shape, number of contractile vacuoles, number and stability of pyrenoids, and ultrastructure of pyrenoids and stigmata. This distinction of species based on morphology was also supported by analyses of rbc L gene sequences. The later strongly showed that each species, C. euchlorum (seven strains), C. elongatum (three strains), and C. capillatum (10 strains), forms a robust clade. Although some morphological differences were noted within different strains of C. euchlorum and C. capillatum, these features were regarded as strain-specific because they were not reflected in the rbc L gene phylogenies. In addition, the rbc L gene trees strongly suggested that C. neglectum and C. kasakii are closely related to each other, consistent with the similarity of the ultrastructure of pyrenoids and stigmata between the two species. However, C. kasakii can be distinguished clearly from C. neglectum by its multiple pyrenoids in the chloroplast and acute anterior and posterior ends in the vegetative cell.     

The ultrastructure of Mychonastes ruminatus gen. et sp. nov., a new member of the Chlorophyceae isolated from brackish water

A new monotypic genus of the Chlorophyceae isolated from brackish water of the Chesapeake Bay, Maryland, U.S.A. is described as Mychonastes ruminatus. The alga is compared with similar members of the Oocystaceae. Based on the presence of a thick ruminate cell wall, the lack of discernible pyrenoids, and other vegetative characters, Mychonastes can be delineated from other green coccoid unicells.

The cellular life cycle of Mychonastes was studied by electron microscopy and compared to published studies of Chlorella. The genera are differentiated by wall and chloroplast structure. Mychonastes cell wall is sculptured and changes during the life cycle from a highly irregular appearance when young, to a less irregular state when mature. Mychonastes chloroplast lacks pyrenoids at all times. Chlorella has a smooth wall and pyrenoids which are apparent just after release from the mother cell until division. Organelles of both genera migrate in a similar pattern throughout the life cycle, with Mychonastes producing two or four autospores, and Chlorella producing four or more (rarely two).     

A GREEN DINOFLAGELLATE WITH CHLOROPHYLLS a and b: MORPHOLOGY,FINE STRUCTURE OF THE CHLOROPLAST AND CHLOROPHYLL COMPOSITION1

A green-colored marine unicell has been grown in unialgal culture and its morphology, chloroplast fine structure, and chlorophyll composition investigated. The organism is typical of dinoflagellates in its shape, flagellation, nucleus, mitochondria, and trichocysts. It is similar to Gymnodinium but possesses fine body scales. Chloroplasts and two kinds of vesicles bounded by double membranes, but no organelles obviously identifiable as nuclei or mitochondria, are associated in ribosome-dense cytoplasm separated by a double membrane from the dinophycean cytoplasm. The chloroplasts are unlike any previously reported for dinoflagellates. Each is enclosed by an envelope consisting of a double membrane. Chloroplast lamellae consist of three appressed thylakoids. Interlamellar pyrenoids are present. Pigment analysis reveals chlorophylls a and b but not chlorophyll c. It seems likely that the organism is an undescribed dinoflagellate containing an endosymbiont with chlorophylls a and b and that the reduction of the endosymbiont nucleus and mitochondria has permitted a more initmate symbiosis.     

CHLOROPLAST DEVELOPMENT AND ULTRESTRUCTURE IN THE FRESHWATER RED ALGA BATRACHOSPERMUM1

Chloroplast development and ultrastructure of the freshwater red alga Batrachospermum moniliforme are described. Chloroplasts develop from proplastids which have a double-membraned chloroplast envelope and a parallel double-membraned outer photo-synthetic lamella. Of these 2 double-membraned structures of the proplastid, only the outermost pho-tosynthetic lamella functions in production of further lamellae. The mature chloroplast consists of 2 or more concentric lamellae and a variable number of nonconcentric lamellae. These lamellae are not dense, uninterrupted sheets as described for other red algae, but are largely constructed of tubules, lying side by side, that form interrupted lamellar sheets. The possible physiological significance of lamellar interruptions in providing path-ways for movement of materials in the chloroplast stroma is discussed.     

Fasciola hepatica: the ultrastructure of the epithelium of the seminal vesicle, the ejaculatory duct and the cirrus.

The ultrastructure of the seminal vesicle, ejaculatory duct, cirrus sac and cirrus is described. The epithelium of the seminal vesicle consists of a single layer of squamous to cuboidal cells. The apical ends of the cells have thin polymorphic lamellae and long narrow pits, both of which enclose normal spermatozoa. The cells have a moderate amount of GER and Golgi complexes which produce a lucid secretory body. The ejaculatory duct epithelium is composed of cuboidal to columnar cells between or through which project the terminal parts of the ducts of the unicellular prostate glands. The apical surfaces of the epithelia are extended into triangular or filiform projections having thin sinuous lamellae. The cytoplasm contains GER cisternae and Golgi complexes which synthesize a dense ovoid secretion. The cirrus sac and cirrus are covered by a thin modified tegument. The cirrus has many spines and the normal ratio of T1 and T2 type of secretory bodies, whereas the cirrus sac has few spines and the T2 type of secretory body predominates over the T1 type. The significance and possible functions of the structures observed in the three tissues are discussed.