Structure and formation of the perforated theca defining the Pelagophyceae (Heterokonta), and three new genera that substantiate the diverse nature of the class.

The pelagophytes, a morphologically diverse class of marine heterokont algae, have been historically united only by DNA sequences. Recently we described a novel perforated theca (PT) encasing cells from the Pelagophyceae and hypothesized it may be the first morphological feature to define the class. Here we consolidate that observation, describing a PT for the first time in an additional seven pelagophyte genera, including three genera new to science. We established clonal cultures of pelagophytes collected from intertidal pools located around Australia, and established phylogenetic trees based on nuclear 18S rDNA and plastid rbcL, psaA, psaB, psbA and psbC gene sequences that led to the discovery of three new species: Wyeophycus julieharrissiae and Chromopallida australis form a distinct lineage along with Ankylochrysis lutea within the Pelagomonadales, while Pituiglomerulus capricornicus is sister genus to Chrysocystis fragilis in the Chrysocystaceae (Sarcinochrysidales). Using fixation by high-pressure freezing for electron microscope observations, a distinctive PT was observed in the three new genera described in this paper, as well as four genera not previously investigated: Chrysoreinhardia, Sargassococcus, Sungminbooa and Andersenia. The mechanism of PT formation is novel, being fabricated from rafts in Golgi-derived vesicles before being inserted into an established PT. Extracellular wall and/or mucilage layers assemble exterior to the PT in most pelagophytes, the materials likewise secreted by Golgi-derived vesicles, though the mechanism of secretion is novel. Secretory vesicles never fuse with the plasma membrane as in classic secretion and deposition, but rather relocate extracellularly beneath the PT and disintegrate, the contents having to pass through the PT prior to wall and/or mucilage synthesis. This study substantiates the diverse nature of pelagophytes, and provides further evidence that the PT is a sound morphological feature to define the Pelagophyceae, with all 14 of the 20 known genera studied to date by TEM possessing a PT.

New pelagophytes show a novel mode of algal colony development and reveal a perforated theca that may define the class.

Pelagophytes (Heterokonta) are a morphologically diverse class of marine algae historically united only by DNA sequences. We established clonal cultures of sand-dwelling pelagophytes collected from intertidal pools around Australia. Phylogenetic trees based on nuclear 18S rDNA and plastid rbcL, psaA, psaB, psbA, and psbC sequences revealed two new genera, Gazia and Glomerochrysis, related to Aureoumbra in a distinct lineage within the Sarcinochrysidaceae (Pelagophyceae). The three new species (Gazia saundersii, Gazia australica, and Glomerochrysis psammophila), along with an Australian strain of Aureoumbra geitleri, are characterized by dominant benthic stages that differ significantly from one another, while occasionally producing classic heterokont zoospores. The benthic stage of Ga. saundersii has a novel development not observed in any other colonial alga, consisting of large, spherical colonies (up to 140 µm in diameter) containing c. 2,500 cells that eventually differentiate and segregate into a large number of daughter colonies that are subsequently liberated. Alternatively, colonies may differentiate into a mass of zoospores that escape and settle to develop into new colonies. In Gl. psammophila, cubic packets of cells form large sticky clusters that bind sand together, while Ga. australica and A. geitleri are unicellular species. Using fixation by high-pressure freezing, a distinctive perforated theca was observed by TEM in all genera of this lineage, and we hypothesize this unique covering may be the first morphological feature to characterize most, if not all, pelagophytes. This study substantiates the diverse nature of sand-dwelling pelagophytes as well as their mechanisms for thriving in a dynamic habitat.

Morphological study of the integument and corporal skeletal muscles of two psammophilous members of Scincidae (Scincus scincus and Eumeces schneideri).

Sand deserts are common biotopes on the earth's surface. Numerous morphological and physiological adaptations have appeared to cope with the peculiar conditions imposed by sandy substrates, such as abrasion, mechanical resistance and the potential low oxygen levels. The psammophilous scincids (Lepidosauria) Scincus scincus and Eumeces schneideri are among those. S. scincus is a species frequently used to study displacement inside a sandy substrate. E. schneideri is a species phylogenetically closely related to S. scincus with a similar lifestyle. The aims of this study focus on the morphology of the integument and the muscular system. Briefly, we describe interspecific differences at the superficial architecture of the scales pattern and the thickness of the integument. We highlight a high cellular turnover rate at the level of the basal germinal layer of the epidermis, which, we suggest, corresponds to an adaptation to cutaneous wear caused by abrasion. We demonstrate the presence of numerous cutaneous holocrine glands whose secretion probably plays a role in the flow of sand along the integument. Several strata of osteoderms strengthen the skin. We characterize the corporal (M. longissimus dorsi and M. rectus abdominus) and caudal muscular fibers using immunohistochemistry, and quantify them using morphometry. The musculature exhibits a high proportion of glycolytic fast fibers that allow rapid burying and are well adapted to this mechanically resistant and oxygen-poor substrate. Oxidative slow fibers are low in abundance, less than 10% in S. scincus, but a little higher in E. schneideri.

A new marine prasinophyte genus alternates between a flagellate and a dominant benthic stage with microrhizoids for adhesion.

Prasinophytes (Chlorophyta) are a diverse, paraphyletic group of planktonic microalgae for which benthic species are largely unknown. Here, we report a sand-dwelling, marine prasinophyte with several novel features observed in clonal cultures established from numerous locations around Australia. The new genus and species, which we name Microrhizoidea pickettheapsiorum (Mamiellophyceae), alternates between a benthic palmelloid colony, where cell division occurs, and a planktonic flagellate. Flagellates are short lived, settle and quickly resorb their flagella, the basal bodies then nucleate novel tubular appendages, termed "microrhizoids", that lack an axoneme and function to anchor benthic cells to the substratum. To our knowledge, microrhizoids have not been observed in any other green alga or protist, are slightly smaller in diameter than flagella, generally contain nine microtubules, are long (3-5 times the length of flagella) and are not encased in scales. Following settlement, cell divisions result in a loose, palmelloid colony, each cell connected to the substratum by two microrhizoids. Flagellates are round to bean-shaped with two long, slightly uneven flagella. Both benthic cells and flagellates, along with their flagella, are encased in thin scales. Phylogenies based on the complete chloroplast genome of Microrhizoidea show that it is clearly a member of the Mamiellophyceae, most closely related to Dolichomastix tenuilepsis. More taxon-rich phylogenetic analyses of the 18S rRNA gene, including metabarcodes from the Tara Oceans and Ocean Sampling Day projects, confidently show the distinctive nature of Microrhizoidea, and that the described biodiversity of the Mamiellophyceae is a fraction of its real biodiversity. The discovery of a largely benthic prasinophyte changes our perspective on this group of algae and, along with the observation of other potential benthic lineages in environmental sequences, illustrates that benthic habitats can be a rich ground for algal biodiscovery.

Psammodinium inclinatum gen. nov. et comb. nov. (=Thecadinium inclinatum Balech) is the closest relative to the toxic dinoflagellate genera Gambierdiscus and Fukuyoa.

The heterotrophic sand-dwelling dinoflagellate Thecadinium inclinatum has been re-examined by light and scanning electron microscopy in order to resolve the discrepancies on its plate pattern from the literature, and to obtain its phylogenetic information single-cell PCR technique has been used. The comparison of morphological and molecular information available for other Thecadinium species confirms the genus is polyphyletic and T. inclinatum seems not related to other representatives of the genus sensu lato. Thus, a new genus and combination for the species, Psammodinium inclinatum gen. nov., comb. nov. is proposed. Cells are heterotrophic and strongly laterally flattened, with sulcal pocket. The revised tabulation is: APC 3' 7" 7c 7s? 5"' 1p 2"" with a long-shank fishhook-shaped apical pore and descending cingulum. The cingulum inclines ventrally and declines on the right lateral side producing an asymmetrical epitheca. The epitheca is much smaller than the hypotheca. The phylogenetic results showed a strong relationship with the autotrophic epiphytic genera Gambierdiscus and Fukuyoa, being closely related with the latter. The Gambierdiscus species typically have a tropical and sub-tropical distribution and produce ciguatoxins, causing thousands of intoxications every year by consumption of contaminated fish. Fukuyoa representatives have a wider distribution including warm and temperate waters, and it has been demonstrated that they are also able to produce ciguatoxins, even though at lower amounts. P. inclinatum, which potential toxicity remains to be determined, represents an interesting independent evolutionary branch that resulted in the loss of chloroplasts, the strong lateral compression and the adaptation to sandy habitats in temperate and cold waters.

The golden paradox - a new heterokont lineage with chloroplasts surrounded by two membranes.

A marine, sand-dwelling, golden-brown alga is described from clonal cultures established from a high intertidal pool in southeastern Australia. This tiny, unicellular species, which we call the "golden paradox" (Chrysoparadoxa australica gen. et sp. nov.), is benthic, surrounded by a multilayered cell wall and attached to the substratum by a complex adhesive plug. Each vegetative cell gives rise to a single, naked zoospore with heterokont flagella that settles and may become briefly amoeboid prior to dividing. Daughter cells are initially amoeboid, then either permanently attach and return to the benthic stage or become motile again prior to final settlement. Two deeply lobed chloroplasts occupy opposite ends of the cell and are surrounded by only two membranes. The outer chloroplast membrane is continuous between the two chloroplasts via the outer membrane of the nuclear envelope. Only two membranes occupy the chloroplast-nucleus interface, the inner membrane of the nuclear envelope and the inner chloroplast membrane. A small pyrenoid is found in each chloroplast and closely abuts the nucleus or protrudes into it. It contains an unusual, membrane-bound inclusion that stains with SYBR green but is unlikely to be a nucleomorph. Phylogenies inferred from a 10-gene concatenated alignment show an early-branching position within the PX clade. The unusual morphological features and phylogenetic position indicate C. australica should be classified as a new class, Chrysoparadoxophyceae. Despite an atypical plastid, exploration of the C. australica transcriptome revealed typical heterokont protein targeting to the plastid.

Spatial and temporal distribution of Gymnorhamphichthys rondoni (Gymnotiformes: Rhamphichthyidae) in a long-term study of an Amazonian terra firme stream, Leticia - Colombia

Weakly electric fishes continually emit electric organ discharges (EOD) as a means of communication and localization of objects in their surroundings. Depending on water conductivity, the amplitude of the electric field generated is known to increase with decreases in electrical conductivity of the water. In Amazonian terra firme streams, water conductivity is extremely low and fluctuates constantly due to local and regional rains. In this context, the space between freely moving weakly electric fishes may be expected to decrease, on average, with an increase in water conductivity. To test this hypothesis, we recorded the positions at rest of the sand-dwelling fish Gymnorhamphichthys rondoni in a terra firme stream for several days in alternating months, over two years. Based on daily nearest neighbor distances among individual fish in a grid, we found a uniform temporal distribution pattern (which was not affected by water conductivity) indicative of site fidelity. Here we highlight the role of other factors that could influence resting site fidelity.(AU)

Los peces eléctricos emiten continuamente descargas con su órgano eléctrico (DOE) para comunicarse y localizar objetos a su alrededor. Dependiendo de la conductividad del agua, la amplitud del campo eléctrico generado aumenta con una disminución en la conductividad eléctrica del agua. En los arroyos de terra firme amazónicos, la conductividad eléctrica del agua es extremadamente baja y fluctua constantemente dependiendo de las lluvias locales y regionales. Teniendo esto en cuenta, esperamos que el espacio entre peces eléctricos débiles con libertad de movimiento disminuya, en promedio, de acuerdo al incremento en la conductividad del agua. Para evaluar esta hipótesis, registramos bajo condiciones naturales por varios dias en meses alternos y durante dos años las posiciones en reposo del pez Gymnorhamphichthys rondoni en un arroyo de terra firme. Con los valores de registro diario de la distancia al vecino mas cercano en peces individuales dentro de una grilla, encontramos un patrón de distribución temporal uniforme (que no fue afectado por la conductividad del agua) e indicativo de una fidelidad por los sitios de reposo. Entonces, nosotros resaltamos aquí el posible papel que otros factores pueden estar jugando para entender la fidelidad por sus sitios de reposo.(AU)

Kraftionema allantoideum, a new genus and family of Ulotrichales (Chlorophyta) adapted for survival in high intertidal pools.

The marine, sand-dwelling green alga Kraftionema allantoideum gen. et sp. nov. is described from clonal cultures established from samples collected in coastal, high intertidal pools from south eastern Australia. The species forms microscopic, uniseriate, unbranched, 6-8 µm wide filaments surrounded by a gelatinous capsule of varying thickness. Filaments are twisted, knotted, and variable in length from 4 to 50 cells in field samples but straighter and much longer in culture, up to 1.5 mm in length. Cell division occurs in several planes, resulting in daughter cells of varying shape, from square to rectangular to triangular, giving rise to gnarled filaments. Mature cells become allantoid, elongate with rounded ends, before dividing one time to form bicells comprised of two domed cells. Adjacent bicells separate from one another and mature filaments appeared as a string of loosely arranged sausages. A massive, single, banded chloroplast covered 3/4 of the wall circumference, and contained a single large pyrenoid encased in a starch envelope that measures 1.5-2.5 µm. Filaments were not adhesive nor did they produce specialized adhesive cells or structures. Reproduction was by fragmentation with all cells capable of producing a new filament. No motile or reproductive cells were observed. Filaments in culture grew equally well in freshwater or marine media, as well as at high salinity, and cells quickly recovered from desiccation. Phylogenetic analysis based on the nuclear-encoded small subunit ribosomal RNA (18S) shows the early branching nature of the Kraftionema lineage among Ulotrichales, warranting its recognition as a family (Kraftionemaceae).

Bispinodinium angelaceum gen. et sp. nov. (Dinophyceae), a new sand-dwelling dinoflagellate from the seafloor off Mageshima Island, Japan(1).

A new athecate dinoflagellate, Bispinodinium angelaceum N. Yamada et Horiguchi gen. et sp. nov., is described from a sand sample collected on the seafloor at a depth of 36 m off Mageshima Island, subtropical Japan. The dinoflagellate is dorsiventrally compressed and axi-symmetric along the sulcus. The morphology resembles that of the genus Amphidinium sensu lato by having a small epicone that is less than one third of the total cell length. However, it has a new type of apical groove, the path of which traces the outline of a magnifying glass. The circular component of this path forms a complete circle in the center of the epicone and the straight "handle" runs from the sulcus to the circular component. Inside the cell, a pair of elongated fibrous structure termed here the "spinoid apparatus" extends from just beneath the circular apical groove to a point near the nucleus. Each of two paired structures consists of at least 10 hyaline fibers and this is a novel structure found in dinoflagellates. Phylogenetic analyses based on the SSU and LSU RNA genes did not show any high bootstrap affinities with currently known athecate dinoflagellates. On the basis of its novel morphological features and molecular signal, we conclude that this dinoflagellate should be described as a new species belonging to a new genus.