LIGHT AND ELECTRON MICROSCOPY OF A PUNCTATE SPECIES OF PYRAMIMONAS,P. MUCIFERA SP. NOV. (PRASINOPHYCEAE)1

Pyramimonas mucifera sp. nov., a punctate species of the genus, is unusual both behaviorally and at the fine structural level. It forms two distinct populations in culture, one benthic and one planktonic. Planktonic forms are more conventional for the genus, but benthic forms are found in loosely packed mucilage, have flagellar rather than ciliary beating of the flagella, and display a higher degree of metaboly. Ultrastructurally this species is unusual in that it has a unique scale complement and the cells contain numerous muciferous vesicles, leaving only small pockets of cytoplasm containing the usual organelles. This species has a 3–1 type flagellar apparatus but has an additional fibrillar band, a 4–3-2–3 microtubular root system and a flexible synistosome. The discovery of a mucilage-producing species of Pyramimonas draws attention to possible links with other prasinophytes (Halosphaera) and green algae of questionable affiliation (Oltmannsiellopsis and Hafniomonas). It also provides a model of the primitive pyramimonad.     

THE ULTRASTRUCTURE OF PYRAMIMONAS PSEUDOPARKEAE SP. NOV. (PRASINOPHYCEAE) FROM SOUTH AFRICA1

The ultrastructure of Pyramimonas pseudoparkeae sp. nov., a member of the class Prasinophyceae occurring in tidal pools along the east, south and west coast of South Africa, is described. The cell surface is covered by three distinctive body scales whilst the flagellar surfaces possess four types of scales. The structure of these scales is described. P. pseudoparkeae resembles Pyramimonas parkeae Norris and Pearson but differs in the structure of the type 2 body scale. The symmetry and ultrastructure of the cell are described with special attention given to the flagellar apparatus. Preliminary information on the life cycle of this species is presented. This new species is compared with other closely related members of the genus Pyramimonas.     

MANTONIELLA IN ANTARCTIC WATERS INCLUDING THE DESCRIPTION OF M. ANTARCTICA SP. NOV. (PRASINOPHYCEAE)1

We report the finding of two species of Mantoniella from the Weddell Sen and Prydz Bay, Antarctica. As well as M. squamata, a cosmopolitan species, we describe M. antarctica sp. nov., the second species in this genus. M. antarctica is covered with two distinctive types of discoid scales. The flagella bear only the smaller of the scale types whereas the cell body has both types, the larger overlying the smaller.     

FLAGELLAR HAIRS IN PRASINOPHYTES (CHLOROPHYTA): ULTRASTRUCTURE AND DISTRIBUTION ON THE FLAGELLAR SURFACE1

The flagellar hair ultrastructure of 16 strains of species of the prasinophycean genera Mantoniella, Mamiella, Pseudoscourfieldia, Nephroselmis, Tetraselmis, Scherffelia, Pterosperma, and Pyraminonas was examined in detail by whole-mount electron microscopy. The flagellar hairs of all genera displayed a high degree of ultrastructural complexity that was completely conserved within each strain. In all strains, flagellar hairs occurred on the sides of the flagella (lateral hairs); in several strains, special flagellar hairs also were found on the flagellar tips (tip hairs; absent in the Chlorodendrales and in Nephroselmis). Two groups of lateral hairs were distinguished: 1) T-hairs (“Tetraselmis-type” flagellar hairs), characterized by a smooth, tubular shaft of ca. 15 nm diameter and an overall length of 0.5–1.3 μm, and 2) P_t-hairs (“Pterosperma-type lateral flagellar hairs”), which were considerably longer (ca. 1.5–5.4 μm), characterized by a thick shaft of ca. 30 nm diameter, which was covered with a layer of regularly spaced small particles of ca. 10 nm diameter. In both groups of flagellar hairs, a strain-specific number of subunits (1–101) in linear arrangement was attached to the distal end of the shaft. Tip hairs were either structurally related to T-hairs (Mamiellales, Pseudoscourfieldia) or represented a separate group, P_t-hairs (“Pterosperma-type flagellar tip hairs”; Pterosperma, Pyramimonas). In four genera (Mantoniella, Mamiella, Pseudoscourfieldia, Nephroselmis), both groups of lateral hairs occurred together on the same cell. Interestingly in these taxa the P_t-hairs were exclusively attached to the shorter immature flagella (no. 2), but, in contrast, in Mantoniella and Pseudoscourfieldia the tip hairs were restricted to the longer mature flagellum (no. 1). Thus, flagella of different developmental status differ in their hair-scale complement. The occurrence, distribution, and ultrastructure of flagellar hairs can be used to identify and classify prasinophytes at all taxonomic levels.     

PHYLOGENETIC ANALYSIS AND GENOME SIZE OF OSTREOCOCCUS TAURI (CHLOROPHYTA, PRASINOPHYCEAE)

Ostreococcus tauri Courties et Chrétiennot-Dinet is the smallest described autotrophic eukaryote dominating the phytoplanktonic assemblage of the marine Mediterranean Thau lagoon (France). Its taxonomic position was partly elucidated from ultrastructure and high-pressure liquid chromatography (HLPC) pigment analysis. The sequence analysis of the 18S rDNA gene of O. tauri measured here is available in EMBL Nucleotide Sequence Database (accession number: Y15814) and allowed to clarify its phylogenetic position. O. tauri belongs to the Prasinophyceae and appears very close to Mantoniella, a typical scaly Prasinophyceae, morphologically very different from the naked and coccoid Ostreococcus. An electrophoretic analysis of O. tauri shows that the nucleus contains 10.20 mbp. This small genome, fragmented into 14 chromosomes ranging in size from 300 to 1500 kbp, confirms the minimalist characteristics of Ostreococcus tauri.     

ULTRASTRUCTURE AND SYSTEMATICS OF TWO NEW FRESHWATER RED CRYPTOMONADS, STOREATULA RHINOSA, SP. NOV. AND PYRENOMONAS OVALIS, SP. NOV.

The ultrastructure and systematics of two red colored freshwater cryptomonads, Storeatula rhinosa , sp. nov. and Pyrenomonas ovalis , sp. nov., are described for the first time. Storeatula, which had been described from marine waters only, has a single inner periplast sheet and a fibrous surface periplast component. Cells lack a furrow but possess a gullet, a bilobed chloroplast connected by a pyrenoid and a nucleomorph located in an indentation of the pyrenoid. This freshwater Storeatula possesses the same general features as the marine species, but it has a contractile vacuole and lacks the lobed chloroplast of S. major. P. ovalis has the generic characteristics described for marine species of Rhodomonas. These characteristics include a short furrow, a deep gullet, square inner periplast plates with beveled corners, a slightly fibrillar surface periplast component, a single chloroplast with two lobes connected by a pyrenoidal bridge and a nucleomorph located in an indentation of the pyrenoid.     

MORPHOLOGY AND MOLECULAR ANALYSES OF THREE TOXIC SPECIES OF GAMBIERDISCUS (DINOPHYCEAE): G. PACIFICUS, SP. NOV., G. AUSTRALES, SP. NOV., AND G. POLYNESIENSIS, SP. NOV.

Three new dinoflagellate species, Gambierdiscus polynesiensis, sp. nov., Gambierdiscus australes, sp. nov., and Gambierdiscus pacificus, sp. nov., are described from scanning electron micrographs. The morphology of the three new Gambierdiscus species is compared with the type species Gambierdiscus toxicus Adachi et Fukuyo 1979, and two other species: Gambierdiscus belizeanus Faust 1995 and Gambierdiscus yasumotoi Holmes 1998. The plate formula is: Po, 3′, 7", 6C, 8S, 5‴, 1p, 2". Culture extracts of these three new species displayed both ciguatoxin- and maitotoxin-like toxicities. The following morphological characteristics differentiated each species. 1) Cells of G. polynesiensis are 68–85 μm long and 64–75 μm wide, and the cell’s surface is smooth. They are identified by a large triangular apical pore plate (Po), a narrow fish-hook opening surrounded by 38 round pores, and a large, broad posterior intercalary plate (1p) wedged between narrow postcingular plates 2‴ and 4‴. Plate 1p occupies 60% of the width of the hypotheca. 2) Cells of G. australes also have a smooth surface and are 76–93 μm long and 65–85 μm wide in dorsoventral depth. They are identified by the broad ellipsoid apical pore plate (Po) surrounded by 31 round pores and a long and narrow 1p plate wedged between postcingular plates 2‴ and 4‴. Plate 1p occupies 30% of the width of the hypotheca. 3) Cells of G. pacificus are 67–77 μm long and 60–76 μm wide in dorsoventral depth, and its surface is smooth. They are identified by the four-sided apical pore plate (Po) surrounded by 30 round pores. A short narrow 1p plate is wedged between the wide postcingular plates 2‴ and 4‴. Plate 1p occupies 20% of the width of the hypotheca. These three newly described species were also characterized by isozyme electrophoresis and DNA sequencing of the D8–D10 region of their large subunit (LSU) rRNA genes. The consistency between species designations based on SEM microscopy and classification inferred from biochemical and genetic heterogeneities was examined among seven isolates of Gambierdiscus. Their classification into four morphospecies was not consistent with groupings inferred from isozyme patterns. Three molecular types could be distinguished based on the comparison of their LSU rDNA sequences. Although G. toxicus TUR was found to be more closely related to G. pacificus, sp. nov. than to other G. toxicus strains, the molecular classification was able to discriminate G. polynesiensis, sp. nov. and G. australes, sp. nov. from G. toxicus. These results suggest the usefulness of the D8–D10 portion of the Gambierdiscus LSU rDNA as a valuable taxonomic marker.     

THE GENUS ACTINOCYCLUS (BACILLARIOPHYCEAE): FRUSTULE MORPHOLOGY OF A. SAGITTULUS SP. NOV. AND TWO RELATED SPECIES1

Actinocyclus octonarius var. tenellus (Bréb.) Hendey, A. actinochilus (Ehr.) Simonsen and A. sagittulus sp. nov. were examined by light and scanning electron microscopy from field samples and/or culture material. Consistent cingulum patterns of a wide valvocopula and one narrow pleura were found. In A. actinochilus, a small third band was found filling the opening of the pleura and is probably present in the other two taxa. The observed variability of the pseudonodulus, especially in A. actinochilus but also in other members of the genus, was confirmed and our results support the literature definition of Actinocyclus as having: 1) a marginal ring of large labiate processes, laterally expanded internally, 2) no external tubes, 3) essentially radial areolation arranged in fascicles, 4) external cribrum, internal foramen, and 5) a pseudonodulus usually present, but may be absent or difficult to detect on individual valves. On the basis of the present study another characteristic should be added; 6) hyaline bands with a wide valvocopula and at least one pleura. This characteristic appears to extend throughout the family Hemidiscaceae, suggesting a close relationship to those Coscinodiscus species with a single marginal row of large labiate processes and zero or one central labiate processes. In addition, Actinocyclus has been noted to have a thin valve overhang extending outside the valvocopula for as much as one-third of its width.     

ULTRASTRUCTURE AND PHYLOGENETIC RELATIONSHIPS OF CHAETOPELTIDALES ORD. NOV. (CHLOROPHYTA,CHLOROPHYCEAE)1

Vegetative cells and zoospores of Hormotilopsis gelatinosa Trainor & Bold, H. tetravacuolaris Arce & Bold, Planophila terrestris Groover & Hofstetter, and Phyllogloea fimbriata (Korchikov) Silva were examined by transmission electron microscopy. All cells had pyrenoids traversed by cytoplasmic channels. Zoospores were quadriflagellate and had essentially cruciate flagellar apparatuses. Scales were present on free-swimming zoospores. These features are essentially identical to those of Chaetopeltis sp. and are dissimilar to those of other described green algae. The new order Chaetopeltidales is created to accommodate the genera Chaetopeltis, Hormotilopsis, Planophila sensu Groover & Hofstetter, Phyllogloea, Dicranochaete, and Schizochlamys, organisms previously scattered among the orders Tetrasporales, Chloro-coccales, Chlorosarcinales, and Chaetophorales. Members of the order are closely related to the ancestral chlorophycean flagellate genus Hafniomonas, may be ancestral with respect to other Chlorophyceae, and may also be closely related to the ulvophycean order Ulotrichales.     

THE RELATIONSHIP BETWEEN PSEUDOSCOURFIELDIA MARINA AND PYCNOCOCCUS PROVASOLII (PRASINOPHYCEAE, CHLOROPHYTA): EVIDENCE FROM 18S rDNA SEQUENCE DATA

18S rDNA sequences were obtained for the prasinophytes Pseudoscourfieldia marina (Throndsen) Manton and Pycnococcus provasolii Guillard. These sequences, along with those of additional prasinophytes and other green algae and land plants, were used for phylogenetic analyses by the neighbor-joining, maximum parsimony, and quartet puzzling methods. Results indicate that Ps. marina and Py. provasolii are closely allied and that both should be included in the Pycnococcaceae. In addition, results of these sequence analyses and additional pigment analysis indicate that the organism previously identified as Ps. marina, isolate CCMP 717, is, in fact, a Nephroselmis sp. or is closely related to that genus.     

ABSOLUTE CONFIGURATION ANALYSIS OF THE FLAGELLAR APPARATUS OF PTEROSPERMA CRISTATUM (PRASINOPHYCEAE) AND CONSIDERATION OF ITS PHYLOGENETIC POSITION1

Pterosperma cristatum Schiller, a member of the Pra-sinophyceae, was examined with light and electron microscopy with special attention on the absolute configuration of flagellar apparatus components and associated structures. This alga is characterized by asymmetrically arranged basal bodies, connecting fibers and microtubular roots. The microtubular root system is homologous with the cruciate root system, the so-called X-2-X-2 root system typical of non-charophycean green algae. Two ducts are associated with microtubular roots. A similar flagellar apparatus and duct system was found in two other prasinophyte genera, Pyramimonas and Halosphaera. The close phylogenetic affinity of these three genera is discussed.     

中国隙蛛亚科蜘蛛一新种(蜘蛛目,暗蛛科)

描述了隙蛛亚科蜘蛛1新种.痕迹隙蛛,新种Coelotes vestigialis sp.nov.(图1～5)正模(♂),副模10(♂)(♂),湖北神农架自然保护区酒壶坪,1998年7月24日至8月8日,周红章、于晓东采.词源:新种根据中突而命名,指中突消失仅留下痕迹.     

CARRAGEENANS FROM AUSTRALIAN REPRESENTATIVES OF THE FAMILY CYSTOCLONIACEAE (GIGARTINALES, RHODOPHYTA), WITH DESCRIPTION OF CALLIBLEPHARIS CELATOSPORA SP. NOV., AND TRANSFER OF AUSTROCLONIUM TO THE FAMILY ARESCHOUGIACEAE

Ten Australian representatives from seven of the 10 genera presently constituting the family Cystocloniaceae have been analyzed for their cell-wall galactans. Included in our survey are the monotypic Australian-endemic genera Austroclonium , Gloiophyllis , Erythronaema , and Stictosporum , one species of Craspedocarpus , three species of Rhodophyllis , and two species of Calliblepharis . As one of the species of the latter genus is endemic to Western Australia and presently undescribed, we illustrate its habit and anatomical features in formally proposing to name it Calliblepharis celatospora Kraft, sp. nov. All the species surveyed essentially produce typical iota (ι)-carrageenans, with the exception of Austroclonium . The sulfated galactans from Austroclonium predominantly contain the repeating units of ι-, alpha (α)-, and 6'-O-methylated ι- and α-carrageenans; whether these exist as discrete polysaccharides or a complex hybrid structure was not resolved. Thus, Austroclonium carrageenans resemble the polysaccharides from Rhabdonia , Areschougia , and Erythroclonium . Although these latter three genera are currently included in the large gigartinalean family Solieriaceae, all produce significantly different carrageenans from Solieria itself and related genera such as Eucheuma , Kappaphycus , Betaphycus , Sarcodiotheca , Agardhiella , Sarconema , and Callophycus . In consideration of these findings, as well as of significant anatomical similarities, we provisionally recommend reestablishment of the family Rhabdoniaceae Kylin (as the family Areschougiaceae J. Agardh) for Rhabdonia , Areschougia , Erythroclonium , and Austroclonium.     

EFFECTS OF CADMIUM ON GROWTH AND SUPEROXIDE DISMUTASE ACTIVITY OF THE MARINE MICROALGA TETRASELMIS GRACILIS (PRASINOPHYCEAE)1

Marine planktonic algae are frequently exposed to metallic contaminants. Because heavy metals can be assimilated and accumulated by algal cells, they can then be transferred to higher trophic levels of food chains. We studied the effects of cadmium on protein production and the growth of the marine prasinophyte Tetraselmis gracilis (Kylin) Butcher. By means of toxicological assays, we estimated the LC₅₀ of cadmium as 3.2 ppm and 1.8 ppm after 48 h and 96 h of exposure to this heavy metal, respectively. The growth curves and survival percentages of cell cultures in the presence of cadmium were determined, and a proportional reduction of both parameters with increasing metal concentrations was found. When chronically exposed to sublethal concentrations of cadmium, T. gracilis contained high levels of superoxide dismutase (SOD) activity, one of the main enzymes of the cell's antioxidant defense mechanism. Under these growth conditions, total SOD activity in crude extracts was increased by 41% (at 1.5 ppm) and 107% (at 3.0 ppm). Assays of SOD activity in nondenaturing polyacrylamide gels also showed a similar induction by cadmium. These results show that cadmium has potentially toxic properties since it significantly inhibited the growth of T. gracilis at low concentrations and promoted the induction of SOD activity, suggestive of an oxidative stress state. Besides being the first report of SOD in T. gracilis, this work describes experimental evidence of SOD induction by cadmium in this species.     

THE FINE STRUCTURE AND SCALE FORMATION OF CHRYSOLEPIDOMONAS DENDROLEPIDOTA GEN. ET. SP. NOV.(CHRYSOLEPIDOMONADACEAE FAM. NOV., CHRYSOPHYCEAE)1

Chrysolepidomonas gen. nov. is described for single-celled monads with two flagella, a single chloroplast, and distinctive canistrate and dendritic scales. The type species, Chrysolepidomonas dendrolepidota sp. nov., is described for the first time. The canistrate scales bear eight “bumps” on the top surface, and the dendriticscales have a tapered base with a quatrifid tip. These organic scales are formed in the Golgi apparatus and storred in a scale reservoir. The scale reservoir is bounded on two sides by the R₁ and R₂ in microtubular roots of the basal apparatus. The cyst (=stomatocyst, statospore) forms endogenously by means of a silica deposition vesicle. The outer cyst surface is smooth, and the pore region is unornamented. Two other organisms bearing canistrate and dendritic scales, previously assigned to the genus Sphaleromants, are transferred to the genus Chrysolepidomonas. They are C.angalica sp. nov. and C. marine(Pienaar) comb. nov. The distinguishing features of Chrysolepidomonas and Sphaleromantis are discussed. A new family, Chrysolepidomonadceae fam. noc., is described for flagellates covered with organic scales.     

THE GENUS PLATOMA (GIGARTINALES,RHODOPHYTA) WITH A DESCRIPTION OF P. ABBOTTIANA SP. NOV.1

Platoma abbottiana (Gymnophlaeaceae; Gigartinales), a new species of benthic red algae, is described from Isla Mejía and is the second member of this genus reported from the Gulf of California, Mexico. Vegetative anatomy and reproductive details of the carpogonial and auxiliary cell branches of the new species are described and illustrated. Geographical distributions of the seven previously described Platoma species are given. Their distinguishing morphological characteristics are compared to those of the new species. The correct time and place of valid publication of the generic name, Platoma, and some of its binomials are discussed.     

FURTHER STUDIES OF PRESUMEDLY PRIMITIVE GREEN ALGAE,INCLUDING THE DESCRIPTION OF PEDINOPHYCEAE CLASS. NOV. AND RESULTOR GEN. NOV.1

The tiny jumping flagellate originally described as Pedinomonas mikron Throndsen was isolated into pure culture from Australian waters and its ultrastructure critically examined. Pedinomonas mikron differs in behavior and in features of the flagellar apparatus from P. minor, the type species from freshwater, and is referred to the new genus Resultor. The two genera are closely related and form the new class Pedinophyceae, which is characterized by features of the flagellar apparatus, mitosis, and cytokinesis. The flagella show the 11/5 orientation otherwise characteristic of Ulvophyceae and Pleurastrophyceae, but they are arranged end to end as in the Chlorophyceae. The flagellar root system is asymmetric and includes a rhizoplast that emerges from the base of one flagellum but subsequently associates with a microtubular root from the second basal body. Mitosis studied previously by Pickett-Heaps and Ott in Pedinomonas is closed, unlike in other green algae, and the spindle is persistent. No phycoplast or phragmoplast is formed during cytokinesis. The eyespot of the Pedinophyceae is located at the opposite end of the cell from the flagella and adjacent to the pyrenoid, as in the most primitive members of the Prasinophyceae. Members of the Pedinophyceae lack prasinoxanthin and Mg 2,4D, characteristic of certain other primitive green algae. The primitive green algae include the classes Prasinophyceae and Pedinophyceae. Micromonadophyceae Mattox et Stewart is considered a synonym of Prasinophyceae. Two new orders are established, Pedinomonadales, containing all known members of the Pedinophyceae, and Scourfieldiales, with the single family Scourfieldiaceae fam. nov. and the single genus Scourfieldia.     

SCHMITZIA EVANESCENS SP. NOV. (RHODOPHYTA,GIGARTINALES) A NEW SPECIES OF CALOSIPHONIACEAE FROM THE NORTHEASTERN NEW ZEALAND COAST1

A new red alga, Schmitzia evanescens sp. nov. (Gigartinales), is described from the northeastern coast of North I., New Zealand. Gametophytes are diminutive, delicate, mucilaginous spring annuals which grow subtidally on cobble substrata. The uniaxial construction and carposporophyte development place this new taxon in the Calosiphoniaceae, a family of only two genera, Calosiphonia H. M. Crouan et P. L. Crouan and Schmitzia P. C. Silva. This is the first report of this family in New Zealand. The new species has vegetative features of both these genera, but is assigned to Schmitzia on the basis of its unusual carposporophyte development in which the gonimoblast initial arises directly from the connecting filament at some distance from the undifferentiated intercalary auxiliary cell. Because recent investigators of other red algal genera with a similar carposporophyte development have questioned the validity of site of gonimoblast initiation as a generic criterion it is suggested that the separation of Schmitzia and Calosiphonia should be re-evaluated. These and several other genera in the Gymnophlaeaceae, Pseudoanemoniaceae, Dumontiaceae and Gloiosiphoniaceae, can be considered, on morphological and reproductive grounds, to be relatively simple and therefore perhaps primitive members of the Rhodophyta.