Four new dinoflagellates

Fragilidium fissile is a new species of this rare genus. It somewhat resemblesF. subglobosum. It differs from the latter in having a slot and a pore in the first apical plate 1′ (the nomenclature of dinoflagellate plate designation follows the Kofoid system). Both species are also distinguishable by noticeable differences in Po, 1″″ and 1‴.Peridinium tyrrhenicum is a small species differing from all the other known species ofPeridinium in its shape, apical channel and several plates, especially some of the sulcal components.Alexandrium foedum somewhat resemblesA. balechii, but it differs from the latter in that its width is greater than its length, and in the characters of all the main sulcal plates. The above listed species were found in a sample from the Tyrrhenian Sea. The fourth species,Alexandrium andersoni, is a small dinoflagellate obtained in coastal waters off Cape Cod. It differs from all the other species of the minutum group in the very typical shapes of both the 6″ and the S. s.Pentapharsodinium daleii Indelicato and Loeblich is transferred toPeridinium.     

The first description of the potentially toxic dinoflagellate, Alexandrium minutum in Hunts Bay, Kingston Harbour, Jamaica

The occurrence and morphology of the potentially toxic dinoflagellate species Alexandrium minutum found for the first time in Jamaica, were examined and described by light and scanning electron microscopy. Classical morphological examinations of whole cells, the thecal plate pattern of intact cells and more importantly the structure of individual thecal plates of squashed cells, were conducted in an attempt to positively identify the species. Characteristics such as a tear-drop shaped apical pore plate with a comma-shaped apical pore and no anterior attachment pore; a narrow sixth precingular plate; a narrow anterior sulcal plate longer than or approximately as long as it is wide; and a posterior sulcal plate wider than long, confirmed the Jamaican species as A. minutum. This dinoflagellate which produces potent neurotoxins responsible for paralytic shellfish poisoning (PSP) in humans in many parts of the World, as well as mass mortality of various marine flora and fauna, was identified in water samples collected during an extensive bloom of the species in the brackish to saline water body of Hunts Bay, an estuarine arm of Kingston Harbour, Jamaica in August 1994. The highest cell concentration was 4.6 × 10⁵ cells l⁻¹, a concentration which far exceeds acceptable concentrations (<10³ cells l⁻¹) of PSP-toxin producing A. minutum in several countries including: Spain and Denmark. No PSP human symptoms were reported during the bloom; however it was accompanied by a large kill of small pelagic fish extending across a third of the bay. Since then, smaller blooms of A. minutum have occurred with the most recent in February and April 2004. Hunts Bay is an important fishing, shrimping and to some extent oyster/mussel collection area and provides an important source of livelihood and food for many fishermen in nearby fishing communities as well as an important source of food for members of other communities. Although there are no known records of human illness due to PSP in Jamaica, the occurrence and blooming in Jamaican waters of this potentially toxic dinoflagellate, is great cause for concern.     

Ultrastructure of Stylodinium Iittorale (Dinophyceae) with special reference to the stalk and apical stalk complex

The ultrastructure of Stylodinium littorale Horiguchi et Chihara, a marine, sand-dwelling coccoid dinoflagel-late, was investigated with special emphasis on its stalk and the apical stalk complex. The dinoflagellate alternates between non-motile and motile cells in its life cycle. The non-motile cell possesses a long and distinct stalk. The stalk, consisting of a main cylindrical part and a holdfast, is firmly attached to a thecal plate (the apical pore plate). A part of its proximal portion is hollow and V-shaped in section. The V-shaped hollow space is underlain by a projection from the apical pore plate. An apical stalk complex is present in the motile cells and consists of a large apical pore plate and mucilaginous material. The apical pore plate is depressed into the cell, but has a narrow central tubular projection. The mucilaginous stalk-building material is stored between this plate and the outer plate membrane. The tubular projection of the apical pore plate corresponds to the apical pore of other dinoflagellates and its lumen is filled with electron-dense material. The structure of the apical stalk complex is compared with the homologous structure in Bysmatrum arenicola, the only other example of an apical stalk complex that has been investigated. A general ultrastructural survey revealed that S. littorale possesses a typical dinoflagellate cellular structure.     

THE RECLASSIFICATION OF PFIESTERIA SHUMWAYAE (DINOPHYCEAE): PSEUDOPFIESTERIA,GEN. NOV.1

Pfiesteria shumwayae Glasgow et Burkholder is assigned to a new genus Pseudopfiesteria gen. nov. Plate tabulation differences between Pfiesteria and Pseudopfiesteria gen. nov. as well as a maximum likelihood phylogenetic analysis based on rDNA sequence data warrant creation of this new genus. The Kofoidian thecal plate formula for the new genus is Po, cp, X, 4′, 1a, 6′′, 6c, PC, 5+s, 5′′′, 0p, 2′′′′. In addition to having six precingular plates, P. shumwayae comb. nov. also has a distinctive diamond or rectangular‐shaped anterior intercalary plate. Both Pfiesteria and Pseudopfiesteria gen. nov. are reassigned to the order Peridiniales based on an apical pore complex (APC) with a canal (X) plate that contacts a symmetrical 1′, four to five sulcal plates, and the conservative hypothecal tabulation of 5′′′, 0p, and 2′′′′. These morphological characters and the life histories of Pfiesteria and Pseudopfiesteria are consistent with placement of both genera in the Peridiniales. Based on the plate tabulations for P. shumwayae, P. piscicida, and the closely related “cryptoperidiniopsoid” and “lucy” groups, the family Pfiesteriaceae is amended to include species with the following tabulation: 4‐5′, 0‐2a, 5‐6′′, 6c, PC, 5+s, 5′′′, 0p, and 2′′′′ as well as an APC containing a pore plate (Po), a closing plate (cp), and an X plate; the tabulation is expanded to increase the number of sulcal plates and to include a new plate, the peduncle cover (PC) plate. Members of the family have typical dinoflagellate life cycles characterized by a biflagellated free‐living motile stage, a varying number of cyst stages, and the absence of multiple amoeboid stages.     

Laboratory culture and taxonomy ofHymenomonas coronata andOchrosphaera verrucosa (Class Prymnesiophyceae) from the Northwest Pacific

Hymenomonas coronata andOchrosphaera verrucosa, both members of the coccolithophorids, Class Prymnesiophyceae, have been studied by means of electron microscopy and with the aid of laboratory culture. Living specimens of these two species were collected in temperate and subtropical regions of Japan, including the Kii Peninsula and the Ryukyu Islands, and unialgal cultures were established in the laboratory. Their life histories are fundamentally identical, and consist of a non-motile vegetative stage that produces motile cells. The vegetative stage is either unicellular, or a packet consisting of a few cells. Both the non-motile cells and the motile cells are covered with two kinds of scales: these are thin scales of unmineralized nature and coccoliths. These two species differ from each other in the shape of the coccoliths and in the presence or absence of visible rudimentary haptonema, and they have been in separate families. The present study reveals that both species are fundamentally identical in the structure and the distribution of major organelles, especially with respect to two opposed pyrenoids which bulge from chloroplasts, each being traversed by two thylakoid bands, and a group of microtubules forming a flagellar root. On the basis of these characteristics, it would appear more logical to place these two species in the same family, namely the Hymenomonadaceae.     

Pyramidodinium spinulosum sp. nov. (Dinophyceae), a sand‐dwelling non‐motile dinoflagellate from the seafloor (36 m deep) off Mageshima Island,Kagoshima, Japan

A new species of benthic marine dinoflagellate, Pyramidodinium spinulosum Horiguchi, Moriya, Pinto & Terada is described from the deep (36 m) seafloor off Mageshima Island, Kagoshima Prefecture, Japan in the subtropical region of the northwest Pacific. The life cycle of the dinoflagellate consists of a dominant, attached, dome‐shaped, vegetative form and short‐lasting, motile cell. Asexual reproduction takes place by the formation of two motile cells within each non‐motile cell. The released motile cells swim only for a short period and transform directly into the dome‐shaped vegetative form. The duration of the cell cycle varies and can be extremely long, ranging 5–38 days under culture conditions. The non‐motile cell is enclosed by a cell wall and its surface is covered with many (80 – 130) spines of various length. The dinoflagellate is photosynthetic and contains many (more than 50) discoidal chloroplasts. Phylogenetic analysis reveals that the dinoflagellate is closely related to the type species of the genus Pyramidodinium, P. atrofuscum which also possesses a dominant, attached, non‐motile form. However, P. spinulosum can be clearly distinguished from P. atrofuscum by the cell shape (dome‐shaped vs. pyramid‐shaped) and surface ornamentation (spines vs. wart‐like processes) of the non‐motile form. Based on these morphological differences together with molecular evidence, it was concluded that this organism from a deep water sand sample should be described as a second species of the genus Pyramidodinium, P. spinulosum.     

Symbiotic and competitive properties of motility mutants of Rhizobium trifolii TA1

Non-motile mutants of Rhizobium trifolii defective in either flagellar synthesis or function were isolated by transposon Tn5 mutagenesis. they were indistinguishable from motile control strains in growth in both laboratory media and in the rhizosphere of clover roots. When each non-motile mutant was grown together with a motile strain in continuous culture, the numbers of motile and non-motile organisms remained in constant proportion, implying that their growth rates were essentially identical. When inoculated separately onto clover roots, the mutants and wildtype did not differ significantly in the number of nodules produced or in nitrogen fixing activity. However, when mixtures of equal numbers of mutant and wild-type cells were inoculated onto clover roots, the motile strain formed approximately five times more nodules than the flagellate or non-flagellate, non-motile mutants, suggesting that motility is a factor in competition for nodule formation.     

Transient expression of lacZ in bombarded unicellular green alga Haematococcus pluvialis

This paper reports for the first time the transient expression of areporter gene, LacZ, in the unicellular green alga Haematococcuspluvialis. By employing the micro-particle bombardment method,motilecells in the exponential phase showed transient expression oflacZ. This was detected in bombarded motile cells undertherupture-disc pressures of 3103 KPa and 4137 KPa.Transient expression of LacZ gene could not be observed in non-motile cells ofthis alga under the same transformation condition. No LacZ background was foundin either the motile cells or the non-motile cells. The study suggests apromising potential of the SV40 promoter and the lacZreporter gene in genetic engineering of unicellular green algae.     

Laciniporus arabicus gen. et sp. nov. (Dinophyceae,Peridiniales), a new thecate,marine, sand‐dwelling dinoflagellate from the northern Indian Ocean (Arabian Sea)1

A new thecate, photosynthetic, sand‐dwelling marine dinoflagellate, Laciniporus arabicus gen. et sp. nov., is described from the subtidal sediments of the Omani coast in the Arabian Sea, northern Indian Ocean, based on detailed morphological and molecular data. Cells of L. arabicus are small (16.2–30.1 μm long and 13.1–23.2 μm wide), dorsoventrally compressed, with a small apical flap‐shaped projection pointing to the left. The thecal plate pattern is distinguished by minute first precingular plate and sulcus, which extends into the epitheca, with large anterior and right sulcal plates. The Kofoidian thecal tabulation is Po, X, 4′, 2a, 7′′, 6c, 6s, 5′′′, 2′′′′. Morphologically, the revealed plate pattern has an affinity to the Peridiniales, and LSU rDNA based phylogenetic analyses placed L. arabicus within the Thoracosphaeraceae, close to calcareous‐cyst producing scrippsielloids, predatory pfiesteriaceans, and photosynthetic freshwater peridinioids Chimonodinium lomnickii and Apocalathium spp. However, the thecal plate arrangement of L. arabicus differs noticeably from any currently described dinoflagellates, and the species stands out from closely related taxa by extensive differences in physiology and ecology.     

Fine structure of the dinoflagellate Aureodinium pigmentosum gen. et sp. nov.

The morphology and fine structure of a small marine dinoflagellate Aureodinium pigmentosum gen. et sp. nov. is described. In the motile state this organism possesses a delicate theca and two typically dinoflagellate flagella. The fine structure is similar in many respects to that of Woloszynskia micra Leadbeater & Dodge, which has already been described in detail. However, the new genus differs from Woloszynskia in having stalked pyrenoids and not having trichocysts. Peridinin is the main xanthophyll pigment. A non-motile athecate phase of the organism is also described.     

UV-B induction of flavonoid biosynthesis in Scots pine (Pinus sylvestris L . ) seedlings

 Cultivation of Scots pine (Pinus sylvestris L.) seedlings under simulated global radiation including the UV-B band (280 – 320 nm; 220 mW m^–2 UV-B_BE) led to increased formation of the diacylated flavonol glucosides 3″,6″-di-p-coumaroyl-astragalin and 3″,6″-di-p-coumaroyl-isoquercitrin in primary and cotyledonary needles, respectively. 3″,6″-Di-p-coumaroyl-astragalin was also the main constitutive diacylated flavonol glucoside in both needle types. This compound predominantly accumulated in primary needles upon UV-B irradiation, and reached concentrations of 2.4 μmol g^–1 fresh weight (fw). Its concentration was only weakly affected in cotyledonary needles. 3″,6″-Di-p-coumaroyl-isoquercitrin was mainly induced in cotyledonary needles with maximum concentrations of 0.8 to 0.9 μmol g^–1 fw, but was virtually unaffected in primary needles under the same irradiation conditions. Pulse labelling with L-(U-¹⁴C)phenylalanine revealed that these metabolites were formed de novo. Phenylalanine ammonia-lyase (EC 4.3.1.5) and chalcone synthase (EC 2.3.1.74) were only slightly induced by the UV-B treatment. The results described here represent the first report on UV-B-induced flavonoid biosynthesis in a conifer species. Received: 5 December 1995 / Accepted: 20 March 1996     

Karyology of a Marine Non-Motile Dinoflagellate, Pyrocystis lunula

Dinoflagellates have a unique and interesting intracellular architecture such as permanently condensed chromosomes throughout the cell cycle. However the study of dinoflagellate chromosomes is not amendable because of the unusually higher number of chromosomes and problems in sample preparation. The species of Pyrocystis spend most of their life cycle as vegetative cyst forms and have been used as experimental organisms for bioluminescence and circadian rhythms. Here, we documented the content of DNA in different life stages and the chromosome karyology in a marine non-motile dinoflagellate Pyrocystis lunula, through light and fluorescent microscopy, serial ultra-thin sectioning, and three dimension (3D) modeling. The DNA content doubles during DNA synthesis and in the end of the cell division two separate daughter cells have the approximately same fluorescent values for the mother cells. Using serial ultra-thin sectioning and 3D modeling, we report the first ultrastructural karyogram. The cells chosen were at the end of karyokinesis. A total of 98 chromosomes were counted and assigned to 49 pairs. In this species, DNA synthesis appears to occur before, or during asexual division and P. lunula lives a diplontic life cycle.     

Durinskia baltica (Dinophyceae), a newly recorded species and genus from China, and its systematics

A freshwater dinoflagellate was identified as Durinskia baltica (Levander) Carty & Cox by morphological characteristics,with the plate formula:Po,x,4',2a,6',5c,4s,5',2'.Durinskia was a newly recorded dinoflagellate genus for China with two anterior intercalary plates and six characteristic precingular plates.Partial sequences of the small and large subunit ribosomal DNA and internal transcribed spacer sequences for the dinoflagellate cells were obtained from field samples.Molecular phylogenetic results indicated Durinskia species could cluster into a monophyletic group,which were distinct from Peridinium species.According to morphological and molecular evidence,it was agreed that the genus Durinskia was separated from the genus Peridinium,which could be a polyphyletic group.In addition,D.baltica was an infrequent diatom-harboring dinoflagellate which was known to possess an endosymbiotic diatom or diatom-like alga.The phylogenetic analyses indicated that D.baltica had a close affinity with Peridiniopsis penardii and P.niei,common freshwater bloom-forming species in China.     

Pyramidodinium atrofuscum gen. et sp. nov. (Dinophyceae), a new marine sand-dwelling coccoid dinoflagellate from tropical waters

A new marine sand‐dwelling coccoid dinoflagellate Pyramidodinium atrofuscum Horiguchi et Sukigara gen. et sp. nov. is described from Jellyfish Lake, Republic of Palau. The dinoflagellate alternates a non‐motile vegetative stage with a motile gymnodinioid stage within its life cycle. The non‐motile stage is dominant in the life cycle and the dinoflagellate reproduces itself by means of the production of two motile cells. The released motile cell swims only for a short period and is directly transformed into the non‐motile cell. The non‐motile cell is sessile, pyramidal in shape, with a single longitudinal ridge and a double transverse ridge. The surface of the cell wall is covered with many processes. The motile cell has a Gymnodinium‐like morphology, but no apical groove is present. An ultrastructural study revealed that the dinoflagellate possesses typical dinoflagellate organelles. Based on the unique morphology of the vegetative non‐motile stage, we propose a new genus Pyramidodinium for this dinoflagellate, with the type species Pyramidodinium atrofuscum Horiguchi et Sukigara, gen. et sp. nov.     

ALEXANDRIUM TAMUTUM SP. NOV. (DINOPHYCEAE): A NEW NONTOXIC SPECIES IN THE GENUS ALEXANDRIUM1

A new species of the dinoflagellate genus Alexandrium, A. tamutum sp. nov., is described based on the results of morphological and phylogenetic studies carried out on strains isolated from two sites in the Mediterranean Sea: the Gulf of Trieste (northern Adriatic Sea) and the Gulf of Naples (central Tyrrhenian Sea). Vegetative cells were examined in LM and SEM, and resting cysts were obtained by crossing strains of opposite mating type. Alexandrium tamutum is a small‐sized species, resembling A. minutum in its small size, the rounded‐elliptical shape and the morphology of its cyst. The main diagnostic character of the new species is a relatively wide and large sixth precingular plate (6″), whereas that of A. minutum is much narrower and smaller. Contrary to A. minutum, A. tamutum strains did not produce paralytic shellfish poisoning toxins. Phylogenies inferred from the nuclear small subunit rDNA and the D1/D2 domains of the large subunit nuclear rDNA of five strains of A. tamutum and numerous strains of other Alexandrium species showed that A. tamutum strains clustered in a well‐supported clade, distinct from A. minutum.     

Life cycle,behavior and morphology of a new tide pool dinoflagellate,Gymnodinium pyrenoidosum sp. nov. (Gymnodiniales,Pyrrhophyta)

A new tide pool dinoflagellate,Gymnodinium pyrenoidosum Horiguchi et Chihara sp. nov. is described from central Japan. It was found to form dense blooms with a characteristic greenish color from April to November. The species exhibits a characteristic diurnal vertical migration and an alternation of a motile with a nonmotile phase, which are dependent on light intensity and tidal movement. Cells of the motile phase are unarmored and relatively small. They have a single, reticulate chloroplast, orange stigma situated near the sulcus and conspicuous pyrenoid in epicone. The alga reproduces itself by means of zoospores which are produced by the bipartition of protoplasm within the parent cell wall during the nonmotile stage which occurs at night. The occurrence of another type of motile cell, termed a macroswarmer, which differs from normal zoospore in size and shape has also been demonstrated.