Genotypic diversity and distribution of Ostreobium quekettii within scleractinian corals

The green filamentous endolithic alga Ostreobium quekettii resides inside skeletons of scleractinian corals in close proximity with their tissue and plays a role in the viability of the coral and its associates. This study examined the distribution and diversity of O. quekettii within scleractinian corals from the Red Sea (Eilat, Gulf of Aqaba), using a molecular phylogenetic marker. The massive coral species Porites lutea and Goniastrea perisi were sampled from a depth range of 6–55 m, and ribulose 1,5-bisphosphate carboxylase large subunit gene (rbcL) DNA sequence of the alga was amplified and analyzed for diversity and distribution of ecological patterns. This work reveals that O. quekettii has at least seven different clades distributed along a depth gradient in the examined scleractinian corals. Among the seven identified clades, four were found only in P. lutea, while the other two clades are found in both P. lutea and G. perisi. Goniastrea perisi colonies at depth of 30 m had a distinct O. quekettii clade that was absent in P. lutea. It is obvious from this study that the green endolithic alga O. quekettii is not a single genotype as previously considered but a complex of genotypes and that this differentiation is of ecological significance.     

Laboratory culture and taxonomy of two species ofDerbesia (Class Chlorophyceae) in Japan

Two species ofDerbesia (Class Chlorophyceae),D. marina andD. tenuissima, have been studied for the purpose of obtaining a better understanding of their morphological details and life histories, using preserved and living specimens as well as laboratory cultures. The life histories of both species were completed in the laboratory, starting from both zoospores and zygotes. Specimens were collected at Asamushi, Aomori-ken, and Shimoda, Shizuoka-ken. Their life history types are fundamentally identical, zoospores giving rise upon germination toa Halicystis-phase, while zygotes grow into aDerbesia-phase. The thallus of theHalicystis-phase which alternates withD. marina is the same as that ofH. ovalis which grows in the northern regions of Japan. On the other hand, the thallus of theHalicystis-phase alternating withD. tenuissima is the same as that ofH. parvula known to occur in the temperate to subtropical regions of Japan. These results coincide with those obtained withD. marina andD. tenuissima in Europe, where the type localities of both species are located. Specimens assignable to these two species were collected at several localities in Japan and, as a result of detailed examination of the morphology, they are believed to be identical with eitherD. marina orD. tenuissima.     

AXILOSPHAERA AND HETEROTETRACYSTIS,NEW CHLOROSPHAERACEAN GENERA FROM TENNESSEE SOIL12

Two new chlorosphaeracean genera were isolated into axenic culture from soil collected in cedar glades in Cedars of Lebanon State Forest, Wilson County, Tennessee. The distinguishing characteristics of the new monotypic genus Axilosphaera include an axile (asymmetric) ckloroplast with at least 1 pyrenoid and Chlamydomonas-type (walled) zoospores. A. vegetata is the type species. Reproduction is by dissociation of daughter cells following vegetative cell division, by zoospores, and by aplanospores. The new polytypic genus Heterotetracystis, comprising 3 species, H. akinetos, H. macrogranulosa, and H. intermedia, is characterized by a parietal chloroplast with at least 1 pyrenoid and walled zoospores with flagella of unequal length. Reproduction is by dissociation of daughter cells following vegetative cell division and by zoospores. H. akinetos is designated as the type species.     

TWO SPECIES OF THE CHLOROPHYTE GENUS OSTREOBIUM FROM SKELETONS OF ATLANTIC AND CARIBBEAN REEF CORALS1,2

Two species of the siphonaceous chlorophyte Ostreobium inhabiting the aragonite skeletons of Atlantic and Caribbean reef corals were studied. Ostreobium quekettii Bornet & Flahault has been previously reported from these locations, but the species is here amended to include filament forms previously described under the name O. reineckei Bornet. Ostreobium constrictum sp. n. is described here for the first time. The 2 sympatric species are distinguished on the basis of filament morphology and chloroplast form.     

The flagellar root system of zoospores of the green algaChlorosarcinopsis (Chlorosarcinales) as compared withChlamydomonas (Volvocales)

The flagellar root system of zoospores in two species ofChlorosarcinopsis (C. minuta andC. spec.) has been studied in detail. The biflagellate zoospores show a cruciate root system, two of the four microtubular roots containing two microtubules, the other two four microtubules. The flagellar apparatus is otherwise identical with that ofChlamydomonas reinhardi as described byRingo (1967). Evidence is presented that the genusChlamydomonas is characterized by a bilateral symmetric root system (4-2-4-2) rather than a system with four equally numbered roots (i.e. 4-4-4-4). It is suggested that a root system with four identical cruciate roots is not present in any biflagellate algal cell. The taxonomic significance of cruciate root systems in green algae is discussed refering to the identical root systems ofChlorosarcinopsis andChlamydomonas.     

Nematophagous fungi: new species of the Lagenidiales endoparasitic on Rhabditis

Two new species of the Lagenidiales endoparasitic on the nematode Rhabditis are reported from Ontario soils. In Myzocytium lenticulare the biflagellate zoospores encyst at any point on the cuticle of the host and penetrate directly through the wall. Sexual reproduction in this species is by oogamy. The antheridial protoplast migrates through a pore in the adjacent wall and fuses with the oogonial protoplast to produce a thick-walled reticulate oospore. In Lagenidium caudatum large biflagellate zoospores encyst at or near the body orifices of the host. Many of the zoospores possess a distinct tail-like appendage. Germination tubes from the encysted zoospores penetrate the body orifices to infect the host. The thallus in this species is distinctive in being composed of narrow, irregular hyphae which fill the host. No sexual stage is known.     

MORPHOLOGICAL AND ECOLOGICAL STUDY OF PHYSODERMA DULICHII

A new species of aquatic Phycomycete, Physoderma dulichii Johns, parasitic on the aquatic sedge Dulichium arundinaceum (L.) Britt., is described from northern Michigan. This parasite infects and kills the upper epidermal cells of the host leaves. Macroscopically, infection by P. dulichii is indicated by striking brown bands with irregular margins, at intervals on the upper surfaces of the leaves. Like other species of Physoderma, this organism's development includes two distinct phases, an epibiotic monocentric phase producing asexual zoospores and an endobiotic polycentric phase bearing thick-walled resting spores that germinate after an extensive period of maturation at low temperature to form zoospores. The morphology and development of the two phases and of resting spore germination are reported in detail. Only the immature leaves of the host are susceptible to infection, which may be initiated by the introduction of mature resting spores, zoospores from germinated resting spores, or zoospores from epibiotic sporangia. Resting-spore zoospores may also produce the endobiotic stage directly. Initiation of infection in nature requires that the terminal cluster of immature leaves on the host plant be submerged, but infection of subsequently formed leaves of emergent culms can be accomplished through the agency of zoospores from epibiotic sporangia on older leaves. The relation of infected stands of hosts to their environment is discussed and the importance of standing water to infection noted. The geographical distribution of the parasite shows correlation with the drainage basins of the Great Lakes, the St. Lawrence River, and the northern Atlantic Coastal Plain     

Quantification of laminarialean zoospores in seawater by real‐time PCR

Laminarialean plants undergo heteromorphic alternation of generations between the macroscopic diploid sporophyte and the microscopic haploid gametophyte. The generations change through the formation and release of asexual or sexual reproductive cells. It is difficult to monitor the release and diffusion of zoospores into the environment. Furthermore, even if zoospores can be detected, species identification remains difficult. This study attempted to develop a sensitive and fast identification method for laminarialean zoospores using quantitative PCR. In addition, we aimed to estimate the density of zoospores in natural seawater. Specific primers for Saccharina japonica and Undaria pinnatifida were designed and used to estimate the quantity of zoospores in seawater. DNA samples (environmental DNA) were collected from seawater once or twice each month for 2 years at the same area, and seasonal variations in the release of zoospores was monitored. The estimated maturation period based on the number of released zoospores in this study was comparable with those of the previously reported maturation periods of S. japonica and U. pinnatifida sporophytes. This supports the validity of our method in estimating zoospore release from laminarialean plants. The method will be a useful tool for ecological studies on these commercially relevant species.     

Dissolution of Dead Corals by Euendolithic Microorganisms Across the Northern Great Barrier Reef (Australia)

Spatial and temporal variabilities in species composition, abundance, distribution, and bioeroding activity of euendolithic microorganisms were investigated in experimental blocks of the massive coral Porites along an inshore–offshore transect across the northern Great Barrier Reef (Australia) over a 3-year period. Inshore reefs showed turbid and eutrophic waters, whereas the offshore reefs were characterized by oligotrophic waters. The euendolithic microorganisms and their ecological characteristics were studied using techniques of microscopy, petrographic sections, and image analysis. Results showed that euendolithic communities found in blocks of coral were mature. These communities were dominated by the chlorophyte Ostreobium quekettii, the cyanobacterium Plectonema terebrans, and fungi. O. quekettii was found to be the principal agent of microbioerosion, responsible for 70–90% of carbonate removal. In the offshore reefs, this oligophotic chlorophyte showed extensive systems of filaments that penetrated deep inside coral skeletons (up to 4.1 mm) eroding as much as 1 kg CaCO₃ eroded m⁻² year⁻¹. The percentage of colonization by euendolithic filaments at the surface of blocks did not vary significantly among sites, while their depths of penetration, especially that of O. quekettii (0.6–4.1 mm), increased significantly and gradually with the distance from the shore. Rates of microbioerosion (0.1–1.4 kg m⁻² after 1 year and 0.2–1.3 kg m⁻² after 3 years of exposure) showed a pattern similar to the one found for the depth of penetration of O. quekettii filaments. Accordingly, oligotrophic reefs had the highest rates of microbioerosion of up to 1.3 kg m⁻² year⁻¹, whereas the development of euendolithic communities in inshore reefs appeared to be limited by turbidity, high sedimentation rates, and low grazing pressure (rates <0.5 kg m⁻² after 3 years). Those results suggest that boring microorganisms, including O. quekettii, have a significant impact on the overall calcium carbonate budget of coral reef ecosystems, which varies according to environmental conditions.     

Comparative ultrastructure of the zoospores of nine species ofNeochloris (Chlorophyta)

Nine species ofNeochloris can be divided into three groups on the basis of comparative ultrastructure of the flagellar apparatus, the cell wall and the pyrenoid of zoospores. In Group I,N. wimmeri andN. minuta, zoospores are thin-walled, pyrenoids are penetrated by stromal channels, and the basal bodies are in the clockwise absolute orientation and connected by the distal and two proximal fibers. In Group II,N. aquatica, N. vigenis, N. terrestris, N. pyenoidosa, andN. pseudostigmatica, zoospores are naked or covered by fuzzy material, pyrenoids are covered by a continuous starch sheath or invaginated by cytoplasmic channels, basal bodies are directly opposed, the distal fiber is differentiated into a ribbed structure at the central region, a striated microtubule-associated component (SMAC) is continuous between opposite two-membered rootlets and connected to the ribbed structure, proximal ends of basal bodies are covered by partial caps, each two-membered rootlet and a basal body are connected by a striated fiber to the X-membered rootlet associated with the opposite basal body, and the basal bodies, when oriented at wide angles, are joined at their proximal ends by core extensions. In Group III,N. pseudoalveolaris andN. cohaerens, zoospores are naked, pyrenoids are traversed by parallel thylakoids, basal bodies are in the counterclockwise absolute orientation and overlapped, and each X-membered rootlet is connected to the end of the opposite basal body by a terminal cap. It is suggested that the genusChlorococcopsis gen. nov. be erected for the Group I species. Group II, which includes the type species,N. aquatica, should be preserved asNeochloris. The group appears to be closely related to the coenobial generaPediastrum, Hydrodictyon, andSorastrum, and to have affinities with the coenocytic generaSphaeroplea andAtractomorpha as well. It is also suggested that the genusParietochloris gen. nov. be erected in thePleurastrophyceae for the species of Group III.     

Observations on chytridiaceous parasites of Phanerogams

Summary A reexamination of Physoderma maculare Wallroth the type species of the genus, on Alisma, confirms Clinton's (1902) account of the production of an epibiotic stage from resting spore zoospores. The latter on mature host tissue may also give rise to the endobiotic stage which bears resting spores. On seedlings, however, resting spore zoospores produce only epibiotic sporangia. The fungus could not successfully infect Sagittaria, a closely related host.Contribution No. 1152     

Carbon and nutrients of indigestible pollen are transferred to zooplankton by chytrid fungi

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CAULOCHYTRIUM PROTOSTELIOIDES SP. NOV., A NEW CHYTRID WITH AERIAL SPORANGIA

Caulochytrium is a unique genus of chytrids characterized by the production of both sessile zoosporangia and aerial sporangiocarps, the latter unknown in other chytrids. The type species, C. gloeosporii, is an obligate parasite on conidia of the fungus Gloeosporium. The newly described species, C. protostelioides, which was discovered first in the British West Indies and then in North Carolina, is an obligate parasite of the dematiaceous fungus Cladosporium. It differs from the type species in microdimensions, smaller number of zoospores per aerial sporangium, lack of sexuality, production of protostelid-like sporangiocarps that do not parasitize the host and which float freely on water, and an unrelated host fungus. The family Caulochytriaceae and genus Caulochytrium are emended.     

Zoospore ultrastructure in species ofTrebouxia andPseudotrebouxia (Chlorophyta)

Zoospore ultrastructure (incl. flagellar apparatus) has been investigated in three species ofTrebouxia (T. glomerata, T. erici, T. pyriformis) and one species ofPseudotrebouxia (P. impressa) using an absolute configuration analysis. Zoospores in all taxa studied are nearly identical in ultrastructure and exhibit a very distinctive disposition of cell organelles: cells are naked, biflagellate and considerably flattened along the plane of flagellar beat, the single contractile vacuole is located anteriorly in the ventral region of the cell, the nucleus is anteriorly to centrally located in the dorsal region of the cell. A single dictyosome is located close to the anterior, ventral edge of the nucleus. The chloroplast occupies a posterior position in the cell and usually has an anterior profile in the left region of the cell. There are two branched mitochondria per cell or a single mitochondrial reticulum with profiles anterior to the nucleus (in the dorsal region of the cell), and posterior to the nucleus. In zoospores ofTrebouxia spp. the posterior mitochondrial profile is associated with a microbody, inP. impressa zoospores the anterior mitochondrial profiles are associated with a microbody. The zoospores contain a distinctive system of three ER-cisternae: one system links to both basal bodies and extends to the nucleus, the other two systems subtend the plasmamembrane on the left and right broad cell surfaces and extend to the posterior region of the cell. The flagellar apparatus is structurally identical to that previously described for zoospores ofFriedmannia israelensis and exhibits basal body displacement by one basal body diameter into the 11/5 o'clock direction, a non-striated distal connecting fiber, a cruciate microtubular root system lacking system I fibers and presence of a single system II fiber which connects the basal bodies with the nucleus and runs parallel to one of the ER-strands. The left flagellar roots (X-roots) are subtended by a complex set of amorphous and striated material that connects each left root with both basal bodies.—This study demonstrates the close systematic relationship between the phycobiontsTrebouxia andPseudotrebouxia and the generaFriedmannia, Pleurastrum, andMicrothamnion and supports recent classification schemes which place all these taxa into a single order separate from otherChlorophyta. Dedicated to Prof. DrElisabeth Tschermak-Woess on the occasion of her 70th birthday.     

Laboratory culture and taxonomy of two species ofPedobesia (Bryopsidales,Chlorophyceae) in Japan

The morphology ofPedobesia lamourouxii andDerbesia ryukyuensis, both collected in Shimoda and the adjacent areas in central Japan, was studied from field specimens and laboratory cultures. Specimens which had the same morphology as EuropeanP. lamourouxii produced stephanokont zoospores which developed into either prostrate filaments or expanded discoidal thalli similar to those described by Feldmann and Codomier (1974) and Feldmannet al. (1975). Erect filament identical with the thallus found in nature developed directly from prostrate filaments. The specimens which had morphology similar to that ofDerbesia ryukyuensis described by Yamada and Tanaka (1938) also produced stephanokont zoospores which developed similarly to those ofP. lamourouxii. This species is, therefore, a member ofPedobesia, and it is made a new combinationP. ryukyuensis (Yamada et Tanaka) Kobara et Chihara, comb. nov.     

COMPARATIVE ULTRASTRUCTURE OF ZOOSPORES WITH PARALLEL BASAL BODIES FROM THE GREEN ALGAE DICTYOCHLORIS FRAGRANS AND BRACTEACOCCUS SP.

The chlorococcalean algae Dictyochloris fragrans and Bracteacoccus sp. produce naked zoospores with two unequal flagella and parallel basal bodies. Ultrastructural features of the flagellar apparatus of these zoospores are basically identical and include a banded distal fiber, two proximal fibers, and four cruciately arranged microtubular rootlets with only one microtubule in each dexter rootlet. In D. fragrans, each proximal fiber is composed of two subfibers, one striated and one nonstriated, and each sinister rootlet is composed of five microtubules (4/1), decreasing to four away from the basal bodies. In Bracteacoccus sp., each proximal fiber is a single unit, the sinister rootlets are four (3/1) or rarely five (4/1) microtubules, and each basal body is associated with an unusual curved structure. The basic features of the flagellar apparatus of the zoospores of these two algae resemble those of Heterochlamydomonas rather than most other chlorococcalean algae that have equal length flagella, basal bodies in the V-shape arrangement, and clockwise absolute orientation. It is proposed that these algae with unequal flagella and parallel basal bodies have a shared common ancestry within the green algae.     

CELL SHAPE AND WALL PATTERN IN RELATION TO CYTOPLASMIC ORGANIZATION IN PEDIASTRUM SIMPLEX

P. simplex is a single-pronged, fenestrated species of Pediastrum. Comparison is made in regard to cell differentiation and structure with P. boryanum, a 2-pronged, unfenestrated species, with emphasis on the origin of cell wall pattern and the regulation of cell shape. The characteristic wall pattern is initiated with the deposition of plaques of wall material of the outer wall layer when zoospores have assembled in the colony. The pattern is postulated to be templated in the plasma membrane. The inner, thicker wall layer is fibrillar and deposited from vesicles derived from the golgi apparatus. In P. simplex 2–4 dictyosomes are present in contrast to the single dictyosome of P. boryanum. The dictyosomes lie at the concave inner face of the nucleus. Blebs of its ribosome-free outer membrane are contributed to the forming face of the golgi apparatus. Parallel microtubules underlie the plasma membrane in the aggregating zoospores and disappear after the initiation of wall formation. The possible role of microtubules and other organelles in the determination of cell shape in Pediastrum is discussed.     

VARIATION IN OCHLOCHAETE HYSTRIX (CHAETOPHORALES. CHLOROPHYYCEAE) STUDIED IN CULTURE1

Six different isolates of Ochlochaete Thwaites ex Harvey have been studied under identical culture conditions. All the isolates show open branching, a character previously ascribed specifically to O. hystrix Thwaites ex Harv. sensu stricto, and all form hairs on rounded cells in the central part of the thallus, a character hitherto attributed only to O. ferox Huber. Consequently, separation of these two entities on the species level is untenable. The plant described by Huber is referred to O. hystrix var. ferox (Huber) var. nov. The type material of Chaetobolus gibbus Rosenvinger is similar to one of the isolates studied and is included in O. hystrix. Quadriflagellate zoospores have been observed in all the isolates, and in one of them also biflagellate swarmers.     

Phototaxis in a new Allomyces

Summary The zoospores of a new species of Allomyces are highly phototactic. An action spectrum of this phototaxis showed a broad peak in the 470 nm to 525 nm region of the spectrum, and the ultrastructure of the zoospores did not reveal any obvious structural basis for this mechanism. The findings are discussed in relation to phototaxis in Chlamydomonas and the rumposome or fibrous body previously found in several other watermold zoospores.