Molecular genetic delineation of Phaeocystis species (Prymnesiophyceae) using coding and non-coding regions of nuclear and plastid genomes

Sequence variation among 22 isolates representing a global distribution of the prymnesiophyte genus Phaeocystis has been compared using nuclear-encoded 18S rRNA genes and two non-coding regions: the ribosomal DNA internal transcribed spacer 1 (ITS1) separating the 18S rRNA and 5.8S rRNA genes and the plastid ribulose-1,5-bisphosphate carboxylase/oxygenase (RUBISCO) spacer flanked by short stretches of the adjacent large and small subunits (rbcL and rbcS). 18S rRNA can only resolve major species complexes. The analysis suggests that an undescribed unicellular Phaeocystis sp. (isolate PLY 559) is a sister taxon to the Mediterranean unicellular Phaeocystis jahnii; this clade branched prior to the divergence of all other Phaeocystis species, including the colonial ones. Little divergence was seen among the multiple isolates sequenced from each colonial species complex. RUBISCO spacer regions are even more highly conserved among closely related colonial Phaeocystis species and are identical in Phaeocystis antarctica, Phaeocystis pouchetii and two warm-temperate strains of Phaeocystis globosa, with a single base substitution in two cold-temperate strains of P. globosa. The RUBISCO spacer sequences from two predominantly unicellular Phaeocystis isolates from the Mediterranean Sea and PLY 559 were clearly different from other Phaeocystis strains. In contrast, ITS1 exhibited substantial inter- and intraspecific sequence divergence and showed more resolution among the taxa. Distinctly different copies of the ITS1 region were found in P. globosa, even among cloned DNA from a single strain, suggesting that it is a species complex and making this region unsuitable for phylogenetic analysis in this species. However, among nine P. antarctica strains, four ITS1 haplotypes could be separated. Using the branching order in the ITS1 tree we have attempted to trace the biogeographic history of the dispersal of strains in Antarctic coastal waters.     

PHYLOGENETIC POSITION OF CRUSTOMASTIX STIGMATICA SP. NOV. AND DOLICHOMASTIX TENUILEPIS IN RELATION TO THE MAMIELLALES (PRASINOPHYCEAE,CHLOROPHYTA)1

A new marine microalga from the Mediterranean Sea, Crustomastix stigmatica Zingone, is investigated by means of LM, SEM, TEM, and pigment and molecular analyses (nuclear‐encoded small subunit [SSU] rDNA and plastid‐encoded rbcL). Pigment and molecular information is also provided for the related species Dolichomastix tenuilepis Throndsen et Zingone. Crustomastix stigmatica has a bean‐shaped cell body 3–5 μm long and 1.5–2.8 μm wide, with two flagella four to five times the body length. The single chloroplast is pale yellow‐green, cup‐shaped, and lacks a pyrenoid. A small bright yellow stigma is located in the mid‐dorsal part of the cell under the chloroplast membrane. An additional accumulation of osmiophilic globules is at times seen in a chloroplast lobe. Cells lack flat scales, whereas three different types of hair‐like scales are present on the flagella. The main pigments of C. stigmatica are those typical of Mamiellales, though siphonein/siphonaxanthin replaces prasinoxanthin and uriolide is absent. The pigment pool of D. tenuilepis is more similar to that of Micromonas pusilla (Butcher) Manton et Parke and of other Mamiellales. The nuclear SSU rDNA phylogeny shows that the inclusion of C. stigmatica and D. tenuilepis in the Mamiellales retains monophyly for the order. The two species form a distinct clade, which is sister to a clade including all the other Mamiellales. Results of rbcL analyses failed to provide phylogenetic information at both the order and species level. No unique morphological or pigment characteristics circumscribe the mamiellalean clade as a whole nor its two daughter clades.     

CHRYSOCHROMULINA BREVITURRITA SP. NOV., A NEW FRESHWATER MEMBER OF THE PRYMNESIOPHYCEAE1

Chrysochromulina breviturrita sp. nov. is described as a new haptonema-bearing member of the freshwater Prymnesiophyceae. It is ca. three times larger than the only other known freshwater member of the genus (C. parva Lackey)and possesses a haptonema only ca. 0.2–0.3 as long as the haptonema of C. parva. Two equal flagella (1.5–2 as long as the haptonema), a large contractile vacuole, two parietal plasties, a single pyremid, nucleus, Chrysolaminarin-like droplets and a food vacuole are features readily observed with the light microscope. Dried and shadoweast whole mounts examined with the electron microscope reveal the cell surface covered with two types of small, delicate scales: outer spined scales with a clubshaped terminus and with the spine anchored to a circular to oval base-plate by 2–6 (7) branching arms; and, spineles oval plate scales with concentric lines and radiating ridges on one face and parallel ridges on the other. C. breviturrita has been found in nine lakes in Ontario (Canada). Pereliminary observations suggest that it is an important component of lake plankton and has probably not been delected in previous work, more because of its fragility than because of its scarcity. Living specimens are a prerequisite for initial identification, since cells frequently rupture and haptonemata are often lost when the organisms are killed with the commonly used fixatives.     

SOME PRYMNESIACEAE (HAPTOPHYTA,PRYMNESIOPHYCEAE) FROM THE MEDITERRANEAN SEA,WITH THE DESCRIPTION OF TWO NEW SPECIES: CHRYSOCHROMULINA LANCEOLATA SP. NOV. AND C. PSEUDOLANCEOLATA SP. NOV.1

Five species belonging to the family Prymnesiaceae (one Prymnesium and four Chrysochromulina) have been identified in cultures obtained from water collected in the Bay of Banyuls‐sur‐Mer (Mediterranean Sea, France) using LM, SEM, and TEM. Two are described as new species, Chrysochromulina lanceolata sp. nov. and C. pseudolanceolata sp. nov. Both species are large and lanceolate with an acute posterior and two anterior arms. They are easily detectable with LM but difficult to distinguish to species level with live cells, without experience. EM reveals two completely different scale patterns in the two species. Cells of C. lanceolata are 21–38 μm long, 7–12 μm wide, and 3–7 μm thick. They possess two subequal flagella (30–51 and 29–44 μm), and the haptonema is shorter than the flagella (23–37 μm). The cell body is covered by plate and spine scales. Cells of C. pseudolanceolata sp. nov. are slightly smaller (15–18 × 6–8 μm) with more rounded extremities, two subequal flagella (19–26 and 17–24 μm), and the haptonema is longer than the flagella (about 35 μm). Three types of plate scales are observed in this species. Other findings are C. alifera Parke et Manton and C. throndsenii Eikrem (a new record for the Mediterranean Sea). Prymnesium faveolatum Fresnel, a new toxic species recently described, is illustrated with both LM and SEM.     

棕囊藻北部湾株的18S rDNA分子鉴定

为研究北部湾棕囊藻(Phaeocystis)藻华的成因,采用PCR克隆了棕囊藻北部湾株核糖体18S r DNA序列。结果表明,棕囊藻北部湾株具有游动单细胞与群体结构两种形态;其18S r DNA序列和NCBI基因库中球形棕囊藻(Phaeocystis globosa)的同源性为99%~100%,在系统进化树上与不同海域来源的球形棕囊藻聚在一大分支上,且与球形棕囊藻间的遗传距离均小于其他种。首次从分子生物学上确定棕囊藻北部湾株为球形棕囊藻。     

PFIESTERIA PISCICIDA GEN. ET SP. NOV. (PFIESTERIACEAE FAM. NOV.), A NEW TOXIC DINOFLAGELLATE WITH A COMPLEX LIFE CYCLE AND BEHAVIOR1

The newly described toxic dinoflagellate Pfiesteria piscicida is a polymorphic and multiphasic species with flagellated, amoeboid, and cyst stages. The species is structurally a heterotroph; however, the flagellated stages can have cleptochloroplasts in large food vacuoles and can temporarily function as mixotrophs. The flagellated stage has a typical mesokaryotic nucleus, and the theca is composed of four membranes, two of which are vesicular and contain thin plates arranged in a Kofoidian series of Po, cp, X, 4′, 1a, 5″, 6c, 4s, 5″′, and 2″″. The plate tabulation is unlike that of any other armored dinoflagellate. Nodules often demark the suture lines underneath the outer membrane, but fixation protocols can influence the detection of plates. Amoeboid benthic stages can be filose to lobose, are thecate, and have a reticulate or spiculate appearance. Amoeboid stages have a eukaryotic nuclear profile and are phagocytic. Cyst stages include a small spherical stage with a honeycomb, reticulate surface and possibly another stage that is elongate and oval to spherical with chrysophyte-like scales that can have long bracts. The species is placed in a new family, Pfiesteriaceae, and the order Dinamoebales is emended.     

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.     

Historical phytoplankton data of the Marsdiep

Published and unpublished data on phytoplankton of the Marsdiep tidal inlet were studied. Most older data, going back to 1897, are based on net-phytoplankton only, the earliest quantitative (Utermöhl) data being from 1965.Phaeocystis sp. bloomed in the Marsdiep after a spring diatom peak, at least as long ago as 1897. Summer or automn peaks ofPhaeocystis sp., frequent now, were also observed in 1898 and 1899. The duration of thePhaeocystis blooms in 1897 to 1899 was shorter than observed after 1978, but longer than in the early 1970s. The recent (1987 to 1989) duration ofPhaeocystis blooms is 2 to 3 times that of 1897–1899. This increase surpasses normal yearly variation and can be related to anthropogenically caused in crease in nutrient concentrations. A number of diatomspecies, at present numerically dominant in the spring peak, are not mentioned as dominant in the earlier periods of observation. They are small and passed through the nets used.Biddulphia sinensis, at present often abundant, is an immigrant in the North Sea since 1903, and for that reason absent from the earliest Marsdiep observations. No clear trend in duration of diatom blooms is apparent during 1965 to 1989. Anthropogenic eutrophication did not affect diatom blooms. Marsdiep records in the literature ofPhaeocystis globosa, P. pouchetii andP. sp. all refer to the same species.     

ULTRASTRUCTURE OF PRYMNESIUM NEMAMETHECUM SP. NOV. (PRYMNESIOPHYCEAE)1

Based on material collected from Cape Town, a new sand-dwelling, marine species of Prymnesium is described. Using light and electron microscopy, Prymnesium nemamethecum sp. nov. has been found to resemble other species of the genus in size, organelle arrangement, and swimming behavior. It differs from other described species in that it has three types of scales, one of which is confined to the region of appendage insertion and forms a sheath of simple plate scales over the haptonema. In addition, the scales constituting the proximal body scale layer(s) are unusual because they are not simple plate scales but are specifically ornamented.     

Preliminary observations on differences of scale morphology at various stages in the life cycle of ‘Apistonema-Syracosphaera’ Sensu von Stosch

A preliminary study of the fine structure of various stages in the life cycle of von Stosch's solate of ‘Syracosphaera carterae’ has been made by means of sections and whole mounts. Protoplast structure appears to be similar throughout the cycle. However, the Apistonema thallus possesses a cell wall composed of numerous layers of closely adpressed rimless unmineralised scales. Two types of asexual swarmers are shown to be covered with one or two layers of somewhat similar scales. One type of swarmer possesses a short haptonema which is covered with small scales. In addition the coccolith-bearing stages are shown to be identical with Hymenomonas carterae sensu Manton and Peterfi (1969) and this isolate has therefore been transferred to H. carterae.     

控制水稻胚乳淀粉合成代谢若干关键酶基因对花后高温的响应表达

以稻米品质温度敏感型的早籼稻品种嘉早935为材料,利用人工气候箱控温试验和实时荧光定量PCR技术,探讨了不同灌浆温度(日均温分别为22和32 ℃)处理下胚乳淀粉分支酶(SBE)、淀粉去分支酶(DBE)和淀粉合酶(SS)的10个同工型基因(sbe1、sbe3、sbe4、pul、isa1、isa2、isa3、Wx、sss1和sss2a)的相对表达量差异及动态变化特征.结果表明: 淀粉合成相关功能基因对水稻灌浆期高温胁迫的响应表达方式存在明显差异,而且因同工型的类型而不同.在高温处理下,sbe1和sbe3的相对表达量显著下降,二者属于SBE类基因中对高温胁迫较敏感的主要同工型;DBE基因中,pul属于高表达的同工型,而且其对高温胁迫响应比isa1、isa2和isa3敏感;在Wx、sss1和sss2a中,sss2a的相对表达量显著低于sss1和Wx, 但sss2a和sss1对高温胁迫响应比Wx敏感,因此二者可能也是高温胁迫对胚乳淀粉结构进行调控的重要位点,尤其在水稻灌浆的中后期发挥重要作用.     

THREE NEW BENTHIC SPECIES OF PROROCENTRUM (DINOPHYCEAE) FROM CARRIE BOW CAY,BELIZE: P. SABULOSUM SP. NOV., P. SCULPTILE SP. NOV., AND P. ARENARIUM SP. NOV.1

Three new benthic, sand-dwelling dinqflagellate species, Prorocentrum sabulosum, Prorocentrum scuptile, and Prorocentrum arenarium, from coral rubble are described from scanning electron micrographs. Species were identified based on shape, size, surface micromorphology, ornamentation of thecal plates, and architecture of the periflagellar area and intercalary band. Cells of P. sabulosum are oval with a cell size of 48–50 μm long and 41–48 μm wide. The areolae are round to oval and numerous (332–450 per valve) and range from 1 to 1.6 μm in size. The periflagellar area of P. sabulosum bears a wide V-shaped depression with a flat ridge and lacks ornamentation; it accommodates six pores: one large flagellar pore, an adjacent smaller auxiliary pore, and four pores of unknown function. The flagellar and auxiliary pores are surrounded by a narrow apical collar. The intercalary band of P. sabulosum is smooth. Prorocentrum sculptile cells are broadly oval, 32–37 nm long, and 30–32 μm wide in valve view with a deep-sculptured apical area. The valves are smooth and are marked with shallow depressions (856–975 per valve). Some of these depressions have a small round opening (0.13 μm in diameter). The periflagellar area is V-shaped with a deeply indented depression; it accommodates the two flagella and a thin angled apical plate. The intercalary band is smooth. Prorocentrum arenarium cells are nearly round in valve view 30–32 μm in diameter. Thecal surface is smooth with scattered kidney-shaped valve poroids (65–73 per valve) and marginal poroids (50–57 per valve). Length and width of poroids are 0.62 μm and 0.36 μm, respectively. The periflagellar area is an unornamented, broad triangle into which a large flagellar pore and a smaller auxiliary pore are fitted. Both flagella, longitudinal and transverse, protrude from the flagellar pore. The intercalary band is smooth. The presence of a peduncle-like structure (2–3 μm long) in P. arenarium was observed situated in the flagellar pore.     

DIMETHYLSULFONIOPROPIONATE STORAGE IN PHAEOCYSTIS (PRYMNESIOPHYCEAE) SECRETORY VESICLES1

Despite the global importance of dimethylsulfoniopropionate (DMSP)/dimethyl sulfide (DMS) and their role in climate regulation, little is known about the mechanisms of their production and storage in Phaeocystis sp., a major contributor of DMS in polar areas. Phaeocystis secretes polymer microgels, by regulated exocytosis, remaining in condensed phase while stored in secretory vesicles ( Chin et al. 2004 ). In secretory cells, vesicles also store small molecules, which are released during exocytosis. Here, we demonstrated that DMSP and DMS were stored in the secretory vesicles of Phaeocystis antarctica G. Karst. They were trapped within a polyanionic gel matrix, which prevented an accurate measurement of their concentration in the absence of a chelating agent such as EDTA. Understanding the production and the export mechanisms of DMSP and DMS into seawater is important because of the impact the cellular and extracellular pools of these highly relevant biogeochemical metabolites have on the environment. The pool of total DMSP in the presence of Phaeocystis may be underestimated by as much as half. Obtaining accurate budget measurements is the first step toward gaining a better understanding of key issues related to the DMS ocean–air interaction and the effect of phytoplankton DMS production on climate change.     

The genus Phyllomyza Fallén (Diptera: Milichiidae) from Japan, with descriptions of four new species

The Japanese species of the genus Phyllomyza Fallén are revised. Four new species, Phyllomyza kanmiyai sp. nov., P. amamiensis sp. nov., P. proceripalpis sp. nov. and P. japonica sp. nov., are described and illustrated. P. securicornis Fallén is recorded from Japan for the first time. A key to the Japanese species of Phyllomyza is presented.     

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 contribution of single and colonial cells of Phaeocystis pouchetii to spring and summer blooms in the north-eastern North Atlantic

Studies of the phytoplankton ecology in different localities in north-Norwegian fjords, the White Sea and the Barents Sea were carried out in spring and early summer to investigate the contribution of single and colonial stages of Phaeocystis pouchetii to phytoplankton abundance. Three different types of flagellated and four colonial cells were observed in all localities. P. pouchetii was rare under the ice of the Barents and White Seas, but their abundance increased rapidly during ice retreat. Single cell C dominated over colonial cell C, often by 50 times or more. The highest share of colonial cells was encountered in April in northern Norwegian fjords, in May in the Barents Sea and in May–June in the White Sea. At times the single cell dominated the total P. pouchetii biomass in Balsfjord (April 1999, 2001) with hardly any colonies present. In the White Sea colonies of P. pouchetii were less abundant than in the other regions. Cell carbon of P. pouchetii colonies appears never to be as dominating in the north-eastern North Atlantic as P. globosa blooms in coastal regions such as the southern North Sea. However, the lobal matrix of P. pouchetii colonies appears to be less solid than that of P. globosa and partly dissolution of the colony matrix during handling and storage of fixes samples induces uncertainty about the absolute numbers of P. pouchetii colonial cell counts. Despite of that, single cells of P. pouchetii seem to dominate significantly over colonial cell biomass at most sites and during some years and in some regions colonial cells seem rare. We speculate that top-down regulation of Phaeocystis spp. blooms possibly determines the ratio between single and colonial cells.     

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

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

MORPHOLOGY,PLOIDY, PIGMENT COMPOSITION,AND GENOME SIZE OF CULTURED STRAINS OF PHAEOCYSTIS (PRYMNESIOPHYCEAE)1

We examined cell morphology, ploidy level, cell size, pigment composition, and genome size in 16 cultured strains of Phaeocystis Lagerheim. Two strains originated from the Antarctic, 3 from the tropical Western Atlantic, and 11 from temperate regions (Eastern Atlantic, English Channel, North Sea, and Mediterranean Sea). Thirteen strains made colonies morphologically similar to P. glo-bosa Scherffel, whereas three never formed colonies under any circumstances. Five-rayed star-like structures with filaments were observed in 11 strains. In several strains, two ploidy levels were observed, one (haploid) linked to flagellates and one (diploid) linked to colonies. Cell size did not appear to be a very good criterion for distinguishing strains since size distributions overlapped. Pigment analysis by reversed-phase-high-performance liquid chroma-tography allowed the strains to be grouped into three clusters that differed from each other mainly by the relative proportions of three carotenoids: fucoxanthin, 19′-hex-anoyloxyfucoxanthin, and diadinoxanthin. All strains contained low levels of 19′-butanoyloxyfucoxanthin. Differences in genome size measured by flow cytometry delimited at least five groups. On the basis of both pigment composition and genome size, six clusters were defined, one corresponding to an Antarctic species (possibly P. antarc-tica), one to P. globosa, and the rest probably to several yet-undescribed species or subspecies. Two main conclusions emerge from this study. First, the taxonomy of the genus Phaeocystis needs to be clarified through a combination of morphological, biochemical, and molecular studies. Second, sexuality is a prevalent phenomenon in Phaeocystis, but controls of the sexual cycle are most likely strain-dependent.     

MORPHOLOGY,MOLECULAR PHYLOGENY AND TAXONOMY OF TWO NEW SPECIES OF PLEODORINA (VOLVOCEAE,CHLOROPHYCEAE)1

The volvocacean genus Pleodorina has been morphologically characterized as having small somatic cells in spheroidal colonies and anisogamous sexual reproduction with sperm packets. In this study we examined two new species that can be assigned to the genus Pleodorina based on morphology: P. starrii H. Nozaki et al. sp. nov. and P. thompsonii F. D. Ott et al. sp. nov. P. starrii was collected from Japan and had 32‐ or 64‐celled colonies with anterior somatic cells and spheroidal individual cellular sheaths that were weakly attached to each other within the colonial envelope. P. thompsonii from Texas (USA) exhibited four or 12 somatic cells in the anterior pole of 16‐ or 32‐celled colonies, respectively, and had a single large pyrenoid in the chloroplast of mature reproductive cells. The chloroplast multigene phylogeny placed P. starrii and P. indica (Iyenger) H. Nozaki in a clade that was robustly separated from the type species P. californica Shaw and P. japonica H. Nozaki. Pleodorina thompsonii was resolved as a basal branch within a large monophyletic group (Eudorina group) composed of Eudorina, Pleodorina and Volvox (excluding section Volvox). Thus, Pleodorina was found among three separate lineages within the Eudorina group in which Eudorina and Volvox were also resolved as nonmonophyletic. The DNA sequences from additional species/strains as well as recognition of morphological attributes that characterize the monophyletic groups within the Eudorina group are needed to construct a natural generic classification within these members of the Volvocaceae.