Zoospore chemotaxis of mangrove thraustochytrids from Hong Kong

Zoospores of mangrove isolates of Schizochytrium mangrovei KF6, KF7, KF12 (three strains), Thraustochytrium striatum KF9 and Ulkenia sp. KF13 were examined for their chemotactic responses to amino acids, carbohydrates, ethanol, and leaf extracts using a capillary root model. Most leaf extracts of mangrove plants and a marsh grass tested were shown to induce moderate chemotactic responses in zoospores of both S. mangrovei KF6 and Ulkenia sp. KF13. Of the remaining amino acids and carbohydrates evaluated, glutamic acid and pectin induced strong attraction in zoospores of S. mangrovei KF6 and Ulkenia sp. KF13, suggesting these are the major components in leaves which may be responsible for the chemotactic response of thraustochytrid zoospores in nature. Zoospores of T. striatum KF9, in general, showed a weak chemotactic response to all the tested compounds and extracts except cellulose, which elicited a moderate response. The ecological significance of the data presented is discussed.     

Über Plastidenstapel beiBotrydiopsis alpina sowie Anlage und Vermehrung der Stigmen bei dieser undHeterococcus (Xanthophyceae)

Under certain conditions inBotrydiopsis alpina stacks of chloroplasts are formed. They consist of up to 8 elements. In contrast to what is known from other algae in zoosporangia of this species and ofHeterococcus caespitosus, stigmata are formed in early developmental stages. They are reproduced together with the chloroplasts, in which they occupy a position at the edge and near the existing or future incision. At the side of the old stigma a new one is formed, and partitioning of the chloroplast between these two leads to their distribution to the daughter chloroplasts. Young daughter cells in the zoosporangia ofBotrydiopsis alpina contain one chloroplast which undergoes a last unequal division giving rise to one astigmate and usually somewhat smaller and to one stigmate chloroplast. In both species the capacity for locomotion may be suppressed, the presumptive zoospores thereby becoming aplanospores. Autospores in the proper sense were not observed. Their development quite generally is different from that of aplanospores (and zoospores), and both types of spores should be distinguished.

Herrn Professor Dr.Lothar Geitler zum 80. Geburtstag in Verehrung gewidmet.     

Observations of the Life Cycle and Movement of the Thraustochytrid Ulkenia amoeboidea (Bahnweg & Sparrow) from the North Sea*

SYNOPSIS Two isolates of the thraustochytrid Ulkenia amoeboidea (Bahnweg & Sparrow) were obtained from the North Sea. In both isolates, the limax ameba liberated from the thallus rapidly assumed a nematode-like shape and movement characteristic of several sporozoan trophozoites. On rounding up, it underwent repeated binary fission and produced biflagellate zoospores.     

Seasonal development of five Cephaleuros species (Trentepohliaceae,Chlorophyta) on the leaves of woody plants and the behaviors of their gametes and zoospores

The infection cycle of five Cephaleuros species – C. aucubae, C. biolophus, C. japonicus, C. microcellularis, and C. virescens – was clarified by investigating seasonal development and gamete and zoospore behavior at study sites in Matsue, Shimane Prefecture, Japan. Fresh thalli or lesions appeared on the leaves of various woody plants from August and continued to develop in general from April to September in the following year. Gametangia and zoosporangia matured on 2‐ and 3‐year‐old leaves. Gametes were released in C. microcellularis samples collected from late March to late May and in samples of the other four species collected from late April to late July. Zoospores were released in C. biolophus, C. japonicus, and C. virescens samples collected from mid‐May to early August. In the present study, gametes did not conjugate but germinated like zoospores to produce new plants. After inoculation with zoospores and gametes that behave as asexual spores, fresh thalli or lesions became evident in 2–5 months following inoculation and gametes and zoospores were produced on the developed thalli or lesions the following year; one cycle of the infection was completed per year in the study area.     

Freeze tolerance of soil chytrids from temperate climates in Australia

Very little is known about the capacity of soil chytrids to withstand freezing in the field. Tolerance to freezing was tested in 21 chytrids isolated from cropping and undisturbed soils in temperate Australia. Samples of thalli grown on peptone–yeast–glucose (PYG) agar were incubated for seven days at −15 °C. Recovery of growth after thawing and transferring to fresh medium at 20 °C indicated survival. All isolates in the Blastocladiales and Spizellomycetales survived freezing in all tests. All isolates in the Chytridiales also survived freezing in some tests. None of the isolates in the Rhizophydiales survived freezing in any of the tests. However, some isolates in the Rhizophydiales recovered growth after freezing if they were grown on PYG agar supplemented with either 1 % sodium chloride or 1 % glycerol prior to freezing. After freezing, the morphology of the thalli of all isolates was observed under LM. In those isolates that recovered growth after transfer to fresh media, mature zoosporangia were observed in the monocentric isolates and resistant sporangia or resting spores in the polycentric isolates. Encysted zoospores in some monocentric isolates also survived freezing. In some of the experiments the freezing and thawing process caused visible structural damage to the thalli. The production of zoospores after freezing and thawing was also used as an indicator of freeze tolerance. The chytrids in this study responded differently to freezing. These data add significantly to our limited knowledge of freeze tolerance in chytrids but leave many questions unanswered.     

Two newhalophytophthora species,H. tartarea andH. masteri,from intertidal decomposing leaves in saltmarsh and mangrove regions

Two new pythiaceous fungi were obtained from decaying leaves submerged in saltmarsh environments of the east coast of the U.S.A., or fringing mangroves in the Bahama Islands, and are described here asHalophytophthora tartarea andH. masteri. The two species have superficially similar zoosporangia whose dehiscence tubes have ragged-looking apices. However, differences in fine structures and development of the dehiscence tube and plug, characteristics of the dehiscence plugs, and presence or absence of vesicles clearly distinguish the two species.Halophytophthora masteri is the only species ofHalophytophthora that has a zoospore release mechanism involving both an extruded plug and a vesicle. Cultural properties concerning growth and asexual reproduction at various salinities and temperatures are also different between the two, probably reflecting adaptation to their respective habitats. Though zoospore release inH. masteri occurs spontaneously from mature zoosporangia, it is remarkably enhanced inH. tartarea and alsoH. masteri by mildly dehydrating mature zoosporangia followed by rewetting with seawater, which suggests a possible relation between the asexual reproduction of these oomycetes and the tidal rhythm in their natural habitats.     

Atkinsiella dubia and its related species

Atkinsiella dubia, isolated from the mantle of abalone (Haliotis sieboldii), is described and illustrated as a new record from Japan. The fungus was also obtained from the gills of swimming crab (Portunus trituberculatus). Six other species of the genusAtkinsiella have hitherto been reported from various aquatic animals. The fungus is distinguished from the other six species by the morphology of its mycelia and the process of zoospore production. The most distinctive feature is that zoospores in the first motile stage ofA. dubia encyst in zoosporangia, unlike the other species. We therefore proposeHalocrusticida gen. nov. (Lagenidiales, Haliphthoraceae) for the other six species ofAtkinsiella. A key to species of the genusHalocrusticida is provided.     

STUDIES ON THE LIFE CYCLE AND TAXONOMY OF LIGNIERA VERRUCOSA

Miller , Charles E. (A. and M. College of Texas, College Station.) Studies on the life cycle and taxonomy of Ligniera verrucosa. Amer. Jour. Bot. 46(10): 725–729. Illus. 1959.—A study of the roots of Veronica persica Poir. and V. hederaefolia L. plants infected with Sorosphaera veronicae Schroeter revealed intracellular cystosori and zoosporangial sori of Ligniera verrucosa. The zoosporangial phase of this species has been heretofore unknown. The plasmodia of L. verrucosa occur in root hairs, and other epidermal and sub-epidermal cells of the roots. Zoosporangial and cystosoral plasmodia are indistinguishable until cleavage has started. It is thought that plasmodia produced during early infection develop into zoosporangia, while those produced later develop into resting spores. Zoospores discharged from zoosporangia may reinfect host cells developing there into zoosporangial or cystosoral plasmodia. No evidence for any sexual process was observed. The spherical zoosporangia making up a single zoosporangial sorus may be interconnected; a single discharge pore may serve to liberate zoospores from different zoosporangia. In the Plasmodiophorales the classical basis for generic distinction has been the arrangement of the resting spores in the sorus. Ligniera, because of the supposedly uncharacteristic nature of its cystosori, has been suggested as a host-variety of Sorosphaera. A comparative study of the cystosori and zoosporangia of Ligniera and Sorosphaera growing in a single host has led to the conclusion that these genera should be considered distinct.     

Development and ultrastructure of the marine,parasitic oomycete,Lagenisma coscinodisci Drebes (Lagenidiales)

The thalli of Lagenisma coscinodisci are coenocytic. In young thalli each nucleus is associated with two procentrioles and four dictyosomes. Before nuclear division two pairs of procentrioles are found on two opposite poles each lying in a pocket of the nuclear membranes. The spindle microtubules radiate from these poles; the nuclear membrane remains completely closed in all divisional stages. The thallus transforms holocarpically into a zoosporangium. The differentiation begins after the breakdown of the host with the formation of a cell wall and with the development of a discharge tube. During these processes peripheral dilated tubules are observed which are in close, local contact with small infoldings of the plasmalemma. The discharge tube develops independent of the point of entry of the infection tube and enlarges by tip growth. There are several light and electron microscopical observations which suggest that the last nuclear divisions in zoosporangia of old cultures are meiotic.They give rise to sexually determined zoomeiospores.     

Early Devonian fungi: a blastocladalean fungus with sexual reproduction

In this paper we describe a fossil fungus–Paleoblastocladia milleri gen. et sp. nov.–from the 400 million-year-old Early Devonian Rhynie chert that shares numerous features with modern zoosporic fungi placed in the order Blastocladiales. The fungus occurs in tufts that arise from stomata or between the cuticle and epidermis of Aglaophyton major axes. Thallus development begins from an irregular bipolar basal cell that produces a system of intramatrical rhizoids and clavate-shaped extramatrical, nonseptate hyphae. These hyphae develop into two types of mature thalli. Sporothalli are characterized by several orders of dichotomous branching and the production of terminal, globose zoosporangia, as well as thick-walled, pitted resting sporangia. On separate dichotomously branched thalli (gametothalli) are terminal chains of two or three gametangia, in which the terminal one is slightly larger. Despite the fact that all of the reproductive organs contain either zoospores or gametes, none show evidence of discharge papillae. The fossil fungus is compared with extant members of the Blastocladiales, and the presence of sexual reproduction is discussed.     

Anaeromyces mucronatus nov. gen., nov. sp. A new strictly anaerobic rumen fungus with polycentric thallus

A new species of strictly anaerobic fungus was isolated from the cow rumen. It is characterized by a polycentric thallus, a polynuclear rhizomycelium, mucronate zoosporangia and uniflagellated zoospores. It is also singular in that the sporocysts do not react to the specific lectins of L-fucose, N-acetyl-D-galactosamine and diacetyl chitobiose. These characteristics justify the creation of a new genus.     

Lipids in the classification of Nocardioides: Reclassification of Arthrobacter simplex (Jensen) lochhead in the genus Nocardioides (Prauser) emend. O'Donnell et al. as Nocardioides simplex comb. nov.

The tubulin proteins of Blastocladiella emersonii have been characterized, and the pool sizes of soluble tubulins measured to evaluate turnover during early development. The axonemal tubulins and soluble tubulin dimers were typical of tubulin proteins from other eukaryotes.[³H]cholchicine binding assays were used to estimate the soluble tubulin pools of zoospores and during early development. The free colchicine-binding pool of tubulin in zoospores represents 1% of the soluble protein. It increases by 49% after encystment (at 30 min), decreases to 21% below the spore level by 50 min, and then increases slowly with growth. Neither deflagellation of zoospores prior to encystment, nor inhibition of axonemal disassembly, alter the postencystment pool increases. Disassembly of cytoskeletal microtubules occurs in either circumstance, but can account for only 54% of the pool increase. It was concluded that (1) the retracted axonemal tubulins are not returned to the soluble pool detected by cholchicine binding and are probably degraded; (2) new microtubules are supplied by the preexisting cytoplasmic pool that expands from disassembly of cytoplasmic microtubules; and (3) that the tubulins of the axonemes and soluble pools may be distinct.     

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.     

A Widely Distributed Thraustochytrid Parasite of Diatoms Isolated from the Arctic Represents a gen. and sp. nov.

A unicellular, heterotrophic, eukaryotic parasite was isolated from nearshore Arctic marine sediment in association with the diatom Pleurosigma sp. The parasite possessed ectoplasmic threads that could penetrate diatom frustules. Healthy and reproducing Pleurosigma cultures would begin to collapse within a week following the introduction of this parasite. The parasite (2–10 μm diameter) could reproduce epibiotically with biflagellate zoospores, as well as binary division inside and outside the diatom host. While the parasite grew, diatom intracellular content disappeared. Evaluation of electron micrographs from co‐cultures revealed the presence of hollow tubular processes and amorphic cells that could transcend the diatom frustule, generally at the girdle band, as well as typical thraustochytrid ultrastructure, such as the presence of bothrosomes. After nucleotide extraction, amplification, and cloning, database queries of DNA revealed closest molecular affinity to environmental thraustochytrid clone sequences. Testing of phylogenetic hypotheses consistently grouped this unknown parasite within the Thraustochytriidae on a distinct branch within the environmental sequence clade Lab19. Reclassification of Arctic high‐throughput sequencing data, with appended reference datasets that included this diatom parasite, indicated that the majority of thraustochytrid sequences, previously binned as unclassifiable stramenopiles, are allied to this new isolate. Based on the combined information acquired from electron microscopy, life history, and phylogenetic testing, this unknown isolate is described as a novel species and genus.     

Blidingia minima var. stolonifera var. nov. (Ulvales, Chlorophyta) from British Columbia: systematics, life history and morphogenesis

Blidingia minima var. stolonifera var. nov. is described from Vancouver, British Columbia. In previous literature this variety may have been confused with B. chadefaudii and B. minima var. minima. The new variety is part of the B. minima species complex in which spores germinate by evacuating the original spore and forming a germ tube. B. minima var. stolonifera is characterized by the development of marginal filaments on the basal disc that have one to five colourless cells and terminate in a cell from which a new disc grows. Colourless cells are devoid of chloroplasts and nuclei, and contain only small remnants of cytoplasm. The runner system is considered a mechanism of vegetative growth and propagation that enables a disc to cover a large amount of substratum before producing erect, unbranched thalli. In field and in culture, B. minima var. stolonifera typically reproduces by means of quadriflagellate zoospores; however, three plants from Vancouver formed biflagellate spores. These germinated poorly and developed into highly irregular, branched thalli.     

Relationship between aggressiveness and zoosporangia viability in Plasmopara halstedii (sunflower downy mildew)

Relationship between aggressiveness and zoosporangia viability was studied in seven Plasmopara halstedii (sunflower downy mildew) isolates of races 100, 300, 304, 314, 710, 704 and 714. Aggressiveness criteria including latent period and sporulation density were analysed on sunflower inbred line showing a high level of quantitative resistance. There were significant differences between pathogen isolates for the two aggressiveness criteria. Viability analyses were performed on oval and spheric zoosporangia. The number of zoospores released from oval zoosporangia was significantly higher than those released from spheric ones. The oval zoosporangia for more aggressive isolates of races 100 and 3xx produced more zoospores than the oval ones for less aggressive isolates of races 7xx. There was a significant correlation between aggressiveness criteria and the number of zoospores released from oval zoosporangia and vice versa for zoospores released from spheric ones. It is concluded that the relationship between aggressiveness and oval zoosporangia viability may be established in P. halstedii.     

Functional necessity of the cytoskeleton during cleavage membrane development and zoosporogenesis in Allomyces macrogynus

Cleavage membrane development and cytokinesis were examined in zoosporangia of Allomyces macrogynus treated with cytoskeletal inhibitors and compared to zoosporogenesis under control conditions. Developing membranes were visualized in living zoosporangia with laser-scanning confocal microscopy using the lipophilic membrane dye FM4-64. Under control conditions, cleavage membranes developed in four discrete stages, ultimately interconnecting to delimit the cytoplasm into polygonal uninucleate domains of near uniform size. Disruption of microtubules did not impede the normal four-stage development of cleavage membranes, and cytokinesis occurred with only minor detectable anomalies, although zoospores lacked flagella. Disruption of actin microfilaments did not inhibit membrane formation but blocked nuclear migration and significantly disrupted membrane alignment and cytoplasmic delimitation. This resulted in masses of membrane that remained primarily in cortical regions of the zoosporangia, as did nuclei, throughout zoosporogenesis. Zoospores formed in the absence of microtubules had only a slightly larger mean diameter than control zoospores, although nearly 50% of spores contained two or more nuclei. Microfilament inhibitor treatments produced spores with substantially larger mean diameters and correspondingly larger numbers of nuclei per spore, with greater than 85% containing three or more nuclei. These results showed that a functional actin microfilament cytoskeleton was required for proper alignment of cleavage elements and cytokinesis in Allomyces zoosporangia while microtubules played a less significant role.     

TWO NEW MEMBERS OF THE BLASTOCLADIACEAE. II. MORPHOGENETIC RESPONSES TO O2 AND CO2

The development of Microallomyces dendroideus and Allomyces reticulatus was studied under various mixtures of O₂, CO₂ and N₂. All thalli grown in broth or agar media in air produce abundant resistant sporangia but rarely produce any zoosporangia. In both fungi, the proportion of zoosporangia borne by the thalli increases as the oxygen level decreases. In addition, increased CO₂ strongly stimulates resistant sporangium germination in M. dendroideus. Neither fungus grows in the absence of oxygen; therefore, they seem to be adapted for rapid multiplication in a microaerophilic niche.     

Development of a transplantation technique of Cystoseira amentacea var. stricta and Cystoseira compressa

On the North-Western Mediterranean coastline, the genus Cystoseira (Fucales) has a key role on upper infralittoral community structure. Cystoseira amentacea var. stricta and C. compressa are the most common superficial Cystoseira species. In this area, the coastline is subjected to a strong anthropogenic pressure which has caused severe Cystoseira population decreases. The present study consists in the development of a cheap and easy to use transplantation technique of C. amentacea var. stricta and C. compressa in order to either restore former natural populations or help their settlement on artificial substrates. The thalli were fixed with epoxy glue in holes made by a drill. The success of the technique was assessed by counting the number of remaining transplanted thalli after 3 weeks, 3 months and 6 months. Results were found encouraging with 75% of survival for both species after 6 months. Moreover fertile fronds were observed on the transplanted thalli showing the harmlessness of the technique.