Diversity of swimming behavior in Pyramimonas (Prasinophyceae)

High‐speed videography and inverted light microscope observations of cultures of several Pyramimonas Schmarda species (Pyramimonas chlorina Sym et Pienaar, Pyramimonas disomata McFadden Hill et Wetherbee, Pyramimonas gelidicola McFadden Wether‐bee et Moestrup, Pyramimonas mantoniae Moestrup et Hill, Pyramimonas melkonianii Sym et Pienaar, Pyramimonas mitra Moestrup et Hill, Pyramimonas moestrupii McFadden, Pyramimonas mucifera Sym et Pienaar, Pyramimonas nephroidea McFadden, Pyramimonas orientalis McFadden Hill et Wetherbee, Pyramimonas parkeae Norris et Pearson, Pyramimonas propulsa Moestrup et Hill) revealed swimming behavior to be much more diverse than originally reported for the genus. This diversity shows gradation between extremes of recurved flagellar beating and non‐biphasic ciliary beating. Comparison with the behavior of presumably more primitive members of the Pyramimonadales (Cymbomonas tetramitformis Schiller and Pterosperma cristatum Schiller) leads to the conclusion that the former condition is primitive and the gradation may assist in reconstructing phylogenetic relationships within the genus Pyramimonas, particularly as it is consistent with phylogenies derived from ultrastructural and molecular data.     

AN ADDITIONAL PUNCTATE SPECIES OF PYRAMIMONAS, P. FORMOSA, SP. NOV., AND ITS IMPACT ON THE SUBGENERA PUNCTATAE AND PYRAMIMONAS (PRASINOPHYCEAE, CHLOROPHYTA)

A new punctate species of Pyramimonas Schmarda from South Africa is described. This species is somewhat anomalous and, like P. chlorina Sym et Pienaar (subgenus Pyramimonas McFadden), has characters that bridge the divide between the subgenera Pyramimonas and Punctatae McFadden. Features of these two species, together with the lack of exclusive character sets for either subgenus derived from other species of these subgenera, lead to the conclusion that Punctatae now should be subsumed formally into the subgenus Pyramimonas.     

Pyramimonas tychotreta, sp. nov. (prasinophyceae), a new marine species from antarctica: light and electron microscopy of the motile stage and notes on growth rates

An undescribed marine prasinophyte, Pyramimonas tychotreta, sp. nov., was isolated from a water sample collected near the ice edge in the Weddell Sea (Antarctica) and is characterized by means of light and electron microscopy. This is the second described Antarctic species in the genus and it possesses a cell ultrastructure typical for members of the subgenus Vestigifera McFadden. The quadriflagellated cells measure 8–12 μm in length and 6–7 μm in width and are equipped with seven types of organic scales that cover the flagella and cell body. The scale floor of the box scales is ornamented by quadrants of parallel striations running perpendicular to one another. The scale floor is further characterized by a number of randomly positioned perforations. The wall of the box scales may be solid or possess up to five perforations. The base of the crown scales is square with rounded corners. It is formed of two crossed ribs, the extremities of which are interconnected by a peripheral rib. Four upright arms, attached to the peripheral rib in positions slightly offset from its junction with the cross ribs, join up with the distal extremity of a central upright strut. Each arm possesses two spines. The limuloid scales are cross‐striped by 10–12 ribs. Some details of the flagellar apparatus are briefly reported. Pyramimonas tychotreta is compared with other species of the genus. Experiments were conducted to study the response of growth rate to variations in temperature and salinity in the clonal culture. The best growth rate (0.45 divisions·24 h ^{? 1}) was found at 4.6° C; growth ceased at temperatures in excess of 12° C. Growth in salinities ranging from 15 to 35 psu was similar, but was arrested at 10 psu. These studies suggest that P. tychotreta as a cold stenotherm and euryhaline taxon. New observations are presented on the geographic distribution of previously described species of Pyramimonas Schmarda from the Northern Foxe Basin, Canada.     

Neue Arten der Gattung Pyramimonas Schmarda

Zusammenfassung Es wird auf eine grosse Formenfülle der Gattung Pyramimonas hingewiesen. Mehrere neue Arten and eine neue Varietät, die in Nordmähren (Tschechoslowakei) gefunden werden, werden erwähnt und abgebildet. Folgende Arten werden beschrieben: Pyramimonas recta, P. fasciata, P. radiata, P. conradii, P. cribrum, P. polychloris, P. rapa, P. pluristigma, P. fabelliformis und die Varietdt P. reticulata var. conica.

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Summary The great number of Pyramimonas-species is discussed. Some new species, found in Nordmähren (Czechoslovakia) are described and pictured. The new species described are: Pyramimonas recta, P. fasciata, P. radiata, P. conradii, P. cribrum, P. polychloris, P. rapa, P. pluristigma, P. fabelliformis and the variety P. reticulata var. conica.

     

Species Composition and Seasonal Dynamics of Density and Biomass of Euglenoids in Peter the Great Bay,Sea of Japan

The species composition and seasonal dynamics of density and biomass of euglenoids in the plankton of Peter the Great Bay were examined. Ten species of euglenoids were found. Of these, Eutreptiella braarudiiThrondsen and Eutreptiella eupharyngeaMoestrup et Norris are new records for the Far Eastern seas of Russia. Sharply pronounced peaks of euglenoid density and biomass occurred in the spring and summer. Increased density and biomass of species of the genera EuglenaEhr. and EutreptiaPerty were observed in polluted waters near sewage outlets.     

The resistance to waterlogging of three Eucalyptus species

Summary Seedlings of Ecalyptus grandis W. Hill ex Maiden were found to be more resistant to waterlogging than those of Eucalyptus robusta Sm. and seedlings of both species were more resistant than those of Eucalyptus saligna Sm. Resistance depended on the stress avoidance mechanism of adventitious root formation. The ethylene-releasing growth substance 2-chloroethylphosphonic acid (ethephon) applied to non-flooded plants was used as a tool to study the role of ethylene in the response of plants to waterlogging. Plant height and total dry weight decreased linearly with plant ethylene content. There was no difference between the species in their response to ethephon.     

Transferring the freshwater dinoflagellate Peridinium wisconsinense (Dinophyceae) to the family Thoracosphaeraceae,with the description of Fusiperidinium gen. nov.

Distinctive spindle‐shaped thecae first described by Samuel Eddy in 1930 and assigned to the genus Peridinium Ehrenberg are commonly reported from freshwater environments in eastern North America. We demonstrate that thecae incubated from cysts of Peridinium wisconsinense Eddy have six cingular plates and a protuberant apical pore complex characteristic of the family Thoracosphaeraceae Schiller 1930 emend. Tangen in Tangen et al . 1982. Small subunit ribosomal DNA (SSU rDNA) and internal transcribed spacer (ITS) sequences confirm the close genetic similarity with Chimonodinium lomnickii (Wo?oszyńska) Craveiro, Calado, Daugbjerg, Gert Hansen & Moestrup and with species recently reassigned to the genus Apocalathium Craveiro, Daugbjerg, Moestrup & Calado that was inferred from previously published LSU rDNA analysis of cysts of P. wisconsinense . Despite sharing identical tabulation with the thoracosphaeracean genera Chimonodinium Craveiro, Calado, Daugbjerg, Gert Hansen & Moestrup and Apocalathium , substantial morphological differences in the morphology of both the thecate and cyst stages of P. wisconsinense led us to reassign this species to the genus Fusiperidinium gen. nov. The phylogenetic position of Fusiperidinium wisconsinense comb. nov., inferred from concatenated data of SSU and LSU sequences, suggests that it evolved from the brackish Scrippsiella lineage, independently of the transition that produced the family Peridiniaceae. Cysts described as Geiselodinium tyonekensis Engelhardt from nonmarine strata from Alaska are apparently identical to the resistant cysts produced by F. wisconsinense . The palynologically‐constrained late Middle Miocene age for the Tyonek Formation provides a minimum age of 11.6 Ma for the evolution of this lineage, coinciding with a rapid glacioeustatic decline in sea level. Our findings also call into question the inclusion of the family Thoracosphaeraceae within the order Peridiniales Haeckel.     

An ultrastructural study of Marsupiomonas pelliculata gen. et sp. nov., a new member of the Pedinophyceae

A new species, Marsupiomonas pelliculata gen. et sp. nov. (Pedinophyceae), is described. A single flagellum emerges from a deep pit with a distinctive thickened margin. The flagellum has rigid fibrillar hairs which are probably formed in the perinuclear space. A short second flagellar basal body lies within the cell close to the basal body of the emergent flagellum and the flagellar root system consists of striated and microtubular roots. There is a distinctive theca covering all but the anterior end of the cell and also a single large bright green chloroplast with an immersed pyrenoid surrounded by a starch shell. The wide salinity tolerance of the species is discussed in relation to its distribution in estuarine and salt marsh habitats. The salient features of the new species—the insertion of the emergent flagellum into a deep pit and the possession of a theca—are also seen in Pedinomonas tenuis, and it is suggested that P. tenuis could be transferred to the new genus Marsupiomonas. The class Pedinophyceae now includes three genera (Pedinomonas, Resultor and Marsupiomonas) and the distinguishing features are discussed.     

Phylogenetic relationships of Sargassum (Sargassaceae, Phaeophyceae) with reference to a taxonomic revision of the section Phyllocystae based on ITS-2 nrDNA sequences

Sequences of the end of the 5.8S gene and the internal transcribed spacer 2 (ITS‐2) of nuclear ribosomal DNA have been determined for 19 species of the brown algal genus Sargassum (Sargassaceae), representing three subgenera and eight sections (sections are in parentheses): Phyllotrichia, Bactrophycus (Teretia, Spongocarpus, Halochloa and Repentia) and Sargassum (Acanthocarpicae, Malacocarpicae, Zygocarpicae) to assess the taxonomic position of the section Phyllocystae traditionally included within the Bactrophycus. The sequence of Myagropsis myagroides (Mertens ex Turner) Fensholt (Sargassaceae) was used as an outgroup. Sequences of ITS‐2 were analyzed using neighbor‐joining, parsimony and maximum likelihood methods. The results showed the existence of three clades in Sargassum, corresponding to the three subgenera. The subgenus Phyllotrichia is positioned near the outgroup. Two robust clades were obtained, one corresponding to the subgenus Bactrophycus and the other to the subgenus Sargassum. Sargassum mcclurei Setchell and Sargassum quinhonense Nguyen, the two Phyllocystae investigated, are close to species belonging to the section Zygocarpicae in the subgenus Sargassum. A transfer of the section Phyllocystae to the subgenus Sargassum is therefore proposed on the basis of molecular data (ITS‐2) and morphological data (receptacles and basal leaf).     

Cryptic species of the subgenus Morphnosoma Lutshnik (Coleoptera: Carabidae; genus Pterostichus) from Japan

Pterostichus thunbergi Morawitz and its related species of Japan are revised based on the structure of membranous parts of their genitalia for the first time, under the name of the thunbergi species group. This species group is monophyletic and is placed in the subgenus Morphnosoma Lutshnik. The monotypic subgenus Moritapterus Berlov, erected for P. thunbergi, is synonymized with Morphnosoma. Comparative study of the endophallus revealed that P. thunbergi michinoku Nakane is a junior synonym of P. habui and that four species should be recognized in this group. Three of them are sympatric with P. thunbergi and one of them, Pterostichus (Morphnosoma) robustistylis, is described as a new species. Molecular phylogeny based on the COI gene (mitochondrial [mt]DNA) supported its independence as a distinct species. The resultant trees of mtDNA and comparative study of morphology revealed that P. thunbergi is probably non‐monophyletic. In the present study we suggest the presence of further cryptic species of Morphnosoma in Japan.     

PHENOTYPIC VARIATION AND GENOTYPIC DIVERSITY IN A PLANKTONIC POPULATION OF THE TOXIGENIC MARINE DINOFLAGELLATE ALEXANDRIUM TAMARENSE (DINOPHYCEAE)1

Multiple clonal isolates from a geographic population of Alexandrium tamarense (M. Lebour) Balech from the North Sea exhibited high genotypic and phenotypic variation. Genetic heterogeneity was such that no clonal lineage was repeatedly sampled according to genotypic markers specified by amplified fragment length polymorphism (AFLP) and microsatellites. Subsampling of genotypic data from both markers showed that ordination of individuals by pair‐wise genetic dissimilarity indices was more reliable by AFLP (482 biallelic loci) than by microsatellites (18 loci). However, resulting patterns of pair‐wise genetic similarities from both markers were significantly correlated (Mantel test P < 0.005). The composition of neurotoxins associated with paralytic shellfish poisoning (PSP) was also highly diverse among these isolates and allowed clustering of toxin phenotypes based on prevalence of individual toxins. Correlation analysis of pair‐wise relatedness of individual clones according to PSP‐toxin profiles and both genotypic characters failed to yield close associations. The expression of allelochemical properties against the cryptophyte Rhodomonas salina (Wis?ouch) D. R. A. Hill et Wetherbee and the predatory dinoflagellate Oxyrrhis marina Dujard. manifested population‐wide variation of responses in the target species, from no visible effect to complete lysis of target cells. Whereas the high genotypic variation indicates high potential for adaptability of the population, we interpret the wide phenotypic variation as evidence for lack of strong selective pressure on respective phenotypic traits at the time the population was sampled. Population markers as applied here may elucidate the ecological significance of respective traits when followed under variable environmental conditions, thereby revealing how variation is maintained within populations.     

LIPID,FATTY ACID,AND STEROL COMPOSITION OF EIGHT SPECIES OF KARENIACEAE (DINOPHYTA): CHEMOTAXONOMY AND PUTATIVE LIPID PHYCOTOXINS1

The lipid class, fatty acid, and sterol composition of eight species of ichthyotoxic marine gymnodinioid dinoflagellate (Karenia, Karlodinium, and Takayama) species was examined. The major lipid class in all species was phospholipid (78%–95%), with low levels of triacylglycerol (TAG; 0%–16%) and free fatty acid (FFA; 1%–11%). The common dinoflagellate polyunsaturated fatty acids (PUFA), octadecapentaenoic acid (OPA 18:5ω3), and docosahexaenoic acid (DHA 22:6ω3), were present in all species in varying amounts (14%–35% and 8%–23%, respectively). The very‐long‐chain PUFA (VLC‐PUFA) 28:7ω6 and 28:8ω3 were present at low levels (<1%), and the ratio of these fatty acids may be a useful chemotaxonomic marker at the species level. The typical dinoflagellate sterol dinosterol was absent from all species tested. A predominance of the 4‐methyl and 4‐desmethyl Δ⁸⁽¹⁴⁾ sterols in all dinoflagellate species included 23‐methyl‐27‐norergosta‐8(14),22‐dien‐3β‐ol (Karenia papilionacea A. J. Haywood et Steid, 59%–66%); 27‐nor‐(24R)‐4α‐methyl‐5α‐ergosta‐8(14),22‐dien‐3β‐ol, brevesterol, (Takayama tasmanica de Salas, Bolch et Hallegraeff 84%, Takayama helix de Salas, Bolch, Botes et Hallegraeff 71%, Karenia brevis (C. C. Davis) G. Hansen et Moestrup 45%, Karlodinium KDSB01 40%, Karenia mikimotoi (Miyake et Kominami ex Oda) G. Hansen et Moestrup 38%); and (24R)‐4α‐methyl‐5α‐ergosta‐8(14),22‐dien‐3β‐ol, gymnodinosterol, (K. mikimotoi 48%, Karenia umbella de Salas, Bolch et Hallegraeff 59%, Karlodinium veneficum (D. L. Ballant.) J. Larsen 71%–83%). In Takayama species, five steroid ketones were identified, including for the first time the 3‐keto form of brevesterol and gymnodinosterol. These results indicate a biochemical link between sterol and steroid ketone biosynthesis, suggesting that selected dinoflagellates can make a significant contribution to ketones in marine sediments. The presence of steroid ketones, specific sterols, and fatty acids, and the ratio of VLC‐PUFA may prove to be a useful chemotaxonomic tool for distinguishing between morphologically similar species. The relative levels of the PUFA, OPA, and DHA, coupled with the potential inhibitory action of Δ⁸⁽¹⁴⁾ sterols, may provide an insight into the ichthyotoxicity of these bloom‐forming dinoflagellates.     

MOLECULAR EVOLUTION OF GLUTAMINE SYNTHETASE II AND III IN THE CHROMALVEOLATES1

Glutamine synthetase (GS) is encoded by three distinct gene families (GSI, GSII, and GSIII) that are broadly distributed among the three domains of life. Previous studies established that GSII and GSIII isoenzymes were expressed in diatoms; however, less is known about the distribution and evolution of the gene families in other chromalveolate lineages. Thus, GSII cDNA sequences were isolated from three cryptophytes (Guillardia theta D. R. A. Hill et Wetherbee, Cryptomonas phaseolus Skuja, and Pyrenomonas helgolandii Santore), and GSIII was sequenced from G. theta. Red algal GSII sequences were obtained from Bangia atropurpurea (Mertens ex Roth) C. Agardh; Compsopogon caeruleus (Balbis ex C. Agardh) Mont.; Flintiella sanguinaria F. D. Ott and Porphyridium aerugineum Geitler; Rhodella violacea (Kornmann) Wehrmeyer and Dixoniella grisea (Geitler) J. L. Scott, S. T. Broadwater, B. D. Saunders, J. P. Thomas et P. W. Gabrielson; and Stylonema alsidii (Zanardini) K. M. Drew. In Bayesian inference and maximum‐likelihood (ML) phylogenetic analyses, chromalveolate GSII sequences formed a weakly supported clade that nested among sequences from glaucophytes, red algae, green algae, and plants. Red algal GSII sequences formed two distinct clades. The largest clade contained representatives from the Cyanidiophytina and Rhodophytina and grouped with plants and green algae. The smaller clade (C. caeruleus, Porphyra yezoensis, and S. alsidii) nested within the chromalveolates, although its placement was unresolved. Chromalveolate GSIII sequences formed a well‐supported clade in Bayesian and ML phylogenies, and mitochondrial transit peptides were identified in many of the sequences. There was strong support for a stramenopile‐haptophyte‐cryptophyte GSIII clade in which the cryptophyte sequence diverged from the deepest node. Overall, the evolutionary history of the GS gene families within the algae is complex with evidence for the presence of orthologous and paralogous sequences, ancient and recent gene duplications, gene losses and replacements, and the potential for both endosymbiotic and lateral gene transfers.     

Serial reconstruction of the mitochondrial reticulum in the antarctic flagellate,Pyramimonas gelidicola (Prasinophyceae,Chlorophyta)

Summary A serial reconstruction ofPyramimonas gelidicola McFadden, Moestrup andWetherbee has revealed a large reticulated mitochondrion branching throughout the cell. The possibility of single mitochondria in other members of thePrasinophyceae and the uniformity of the morphology of this organelle within the class is discussed.     

Phylogenetic relationships among species of Prunus as inferred by isozyme markers

Summary An isozyme survey of 34 species of Prunus representing subgenera Prunus, Amygdalus, Cerasus, and Lithocerasus detected 110 presumptive alleles at 11 isozyme loci. Principal component analysis was conducted on the covariance matrix derived from allelic frequencies calculated for each species. Cluster analysis was performed on the first 30 principal components. Results generally support traditional classification of Prunus at the subgeneric level, except for members of subgenus Lithocerasus and two members of subgenus Amygdalus. Prunus glandulosa Thunb., P. japonica Thunb., and P. tomentosa Thunb. of subgenus Lithocerasus and P. triloba Lindl. of subgenus Amygdalus appear to represent primitive species. P. besseyi Bailey and P. pumila L. of subgenus Lithocerasus and P. andersonii of subgenus Amygdalus should be assigned to subgenus Prunus. Placement of its members indicates that subgenus Lithocerasus is an artificial grouping of species that are very different genetically although similar phenotypically.Paper No. 12529 of the Journal Series of the North Carolina Agricultural Research Service, Raleigh, NC 27695-7643, USA     

我国沿海拟菱形藻属的2新记录种及其产毒特征分析

为澄清我国沿海拟菱形藻属(Pseudo-nitzschia)的物种多样性,并确认中国海域拟菱形藻属是否具有产生多莫酸(Domoicacid)的能力,采用毛细管显微操作技术从我国沿海水体中分离、纯化拟菱形藻细胞,建立了单克隆培养株系,并基于核糖体转录间隔区ITS1-5.8S-ITS2 序列构建了分子系统树。结果表明,结合在光学显微镜和透射电镜下观察的形态学特征和分子系统发育分析数据,鉴定到我国拟菱形藻属的2新记录种:银河拟菱形藻(P. galaxiae Lundholm & Moestrup)和微孔拟菱形藻(P. micropora Priisholm, Moestrup & Lundholm),对其形态学特征进行了详细描述,并与相似种类进行了比较研究。利用高效液相色谱(HPLC)技术对多莫酸特征进行了检测,结果表明培养株系并不产生多莫酸。这些为我国拟菱形藻属物种多样性和产毒特征研究提供了基础数据。     

Testing systematic concepts of Sargassum (Fucales, Phaeophyceae) using portions of the rbcLS operon

The taxonomic and phylogenetic concepts within the Sargassum C. Agardh (Sargassaceae) species complex were evaluated through molecular phylogenetic analyses using portions of the chloroplast encoded rbcLS Operon. According to more conservative sequences (rbcL), Turbinaria (Turner) J. Agardh is a close and well‐supported sister lineage to the Sargassum species complex and an appropriate external outgroup for analyses of subgenera and subsections within Sargassum. Both rbcL and more rapidly evolving rbcLS spacer sequences indicated that the East Asiatic genus Myagropsis (Mertens et Turner) Fensholt, along with Sargassum sinicola Setchell et Gardner, represent the closest lineage to Sargassum and form appropriate internal outgroups. The rbcLS spacer region supported three of four subgeneric designations by J. Agardh and sectional levels within the subgenus Sargassum. However, some aspects of Agardh's system were not supported: many of the subsectional ranks or the phyletic concepts; Phyllotrichia was not monophyletic as a subgenus, and its species were also not the most ancestral of Sargassum; and subgenus Sargassum was not the most derived subgenus within the genus. This modern phylogeny suggests a deep evolutionary history for subgenus Sargassum with rapid speciation in closely related subsections and series, and a sister relationship between subgenera Arthrophycus and Bactrophycus.