A novel milky disease organism from Australian scarabaeids: Field occurrence,isolation, and infectivity

A novel milky disease organism has been found causing disease in Aphodius tasmaniae and other scarabaeid larvae in the field in Australia. The sporangium is exceptionally long, measuring 10.5 × 1.5 μm, with a small central spore, measuring 1.0 × 0.6 μm. The vegetative cell is about half the size of the sporangium. The disease was easily transmitted by injection of spores into the hemocoel, with typically milky symptoms developing in 2–4 weeks. Spores will form in vivo at temperatures down to 12°C. For A. tasmaniae third-instar larvae, the ID₅₀ by injection was 3 × 10² spores/larva, yet no infection resulted when larvae were reared in peat containing up to 10⁸ spores/g, i.e., the disease was not successfully transmitted per os. All 10 species of scarabaeids tested were susceptible to the disease when spores were injected; however, all attempts to infect larvae per os were unsuccessful. In vitro culture was also unsuccessful.     

Systematics of the genus Ceratopteris Brongn. (Parkeriaceae) II. Taxonomy

A revision of the genusCeratopteris is presented based on comparative morphology and geographical distribution. Four species are recognized:C. thalictroides, C. cornuta, C. pteridoides, andC. richardii. Characteristics of taxonomic importance include frond length, shape, and dissection; stipe width; insertion of basal pinnae; bud development; habit; annulus cell number; spore number per sporangium; spore size; and spore surface features. Evolution within species and putative hybridization between species are discussed.     

Sporangium Exposure and Spore Release in the Peruvian Maidenhair Fern (Adiantum peruvianum,Pteridaceae)

We investigated the different processes involved in spore liberation in the polypod fern Adiantum peruvianum (Pteridaceae). Sporangia are being produced on the undersides of so-called false indusia, which are situated at the abaxial surface of the pinnule margins, and become exposed by a desiccation-induced movement of these pinnule flaps. The complex folding kinematics and functional morphology of false indusia are being described, and we discuss scenarios of movement initiation and passive hydraulic actuation of these structures. High-speed cinematography allowed for analyses of fast sporangium motion and for tracking ejected spores. Separation and liberation of spores from the sporangia are induced by relaxation of the annulus (the ‘throwing arm’ of the sporangium catapult) and conservation of momentum generated during this process, which leads to sporangium bouncing. The ultra-lightweight spores travel through air with a maximum velocity of ~5 m s^-1, and a launch acceleration of ~6300g is measured. In some cases, the whole sporangium, or parts of it, together with contained spores break away from the false indusium and are shed as a whole. Also, spores can stick together and form spore clumps. Both findings are discussed in the context of wind dispersal.     

MORPHOLOGY AND LABORATORY CULTIVATION OF ECHINOSTELIUM MINUTUM

Alexopoulos , Constantine J. (State U. Iowa, Iowa City.) Morphology and laboratory cultivation of Echinostelium minutum. Amer. Jour. Bot. 47(1): 37—43. Illus. 1960.—The morphology of the sporangium, spores, swarm cells and Plasmodium of the white form of Echinostelium minutum is described. A peridium is present in the early stages of sporangial formation. It eventually disappears leaving only a small collar at the base of the columella. The structure of the spore wall is unique in this genus. The spore case may be described as consisting of a thin wall with several thickened portions distributed over its surface. These are particularly evident in germinated spores. Spore germination and swarm cells are described for the first time. Swarm cells are biflagellate with two long anterior flagella of nearly equal length. The Plasmodium remains microscopic until fruiting time, when it gives rise to but a single sporangium. The plasmodial protoplast never becomes differentiated into veins but remains more or less homogeneous. It exhibits almost imperceptibly slow, irregular streaming instead of the reversible, rapid, rhythmic motion characteristic of plasmodia of most other Myxomycetes which have been studied. It typifies, therefore, a third type of Plasmodium which may be placed alongside that of the Physarales, and that of Stemonitis flavogenita. The laboratory cultivation of E. minutum from spore to spore on agar media is reported here for the first time.     

CotC-CotU heterodimerization during assembly of the Bacillus subtilis spore coat

We report evidence that CotC and CotU, two previously identified components of the Bacillus subtilis spore coat, are produced concurrently in the mother cell chamber of the sporulating cell under the control of σ^K and GerE and immediately assembled around the forming spore. In the coat, the two proteins interact to form a coat component of 23 kDa. The CotU-CotC interaction was not detected in two heterologous hosts, suggesting that it occurs only in B. subtilis. Monomeric forms of both CotU and CotC failed to be assembled at the surface of the developing spore and accumulated in the mother cell compartment of cells mutant for cotE. In contrast, neither CotU nor CotC accumulated in the mother cell compartment of cells mutant for cotH. These results suggest that CotH is required to protect both CotU and CotC in the mother cell compartment of the sporangium and that CotE is needed to allow their assembly and subsequent interaction at the spore surface.     

REPRODUCTION IN ZONARIA FARLOWII. I. GAMETOGENESIS,SPOROGENESIS, AND EMBRYOLOGY1

The brown alga Zonaria farlowii which exhibits an alternation of isomorphic generations has been found to be a particularly suitable organism for a comparative study of game to gene sis, sporogenesis, and embryonic development. The germinations of the zygote and spore have been compared and found to be similar. The development of the oogonium and sporangium has been studied with the light and electron microscopes, in an attempt to discern ontogcnetical similarities or differences. The timing and periodicity of oogenesis and sporogenesis were found to differ: oogonial development requires a lunar month from the time of its initiation to the time of egg release which occurs around the first quarter of the moon, whereas the sporangium does not follow this periodicity. The development of the spore embryos and zygotic embryos is essentially the same. It appears that the difference between the 2 phases of the life cycle exists only in the later stages of oogenesis and sporogenesis. These differences continue in the egg until fertilization, after which the zygote has the same cytoplasmic configuration as the spore. It is also notable that during release of the spore it must increase in size considerably since before germination begins it has attained the size of the egg.     

A segregated model for heterologous amylase production by Bacillus subtilis

A segregated model was proposed to investigate the inherent relationships between growth, substrate consumption, cell differentiation and product formation in a Bacillus subtilis fermentation producing heterologous amylase in a 22-l bioreactor. The segregated model includes three distinguishable cell states and the transition from vegetative phase to sporangium and finally to mature spore. An age-based population balance model was applied to describe the maturity of sporangium toward the formation of spores. Parameters in the model were determined by fitting the model with experimental data. The model was able to predict the transient behavior of B. subtilis in both batch and fed-batch cultures.     

Spiroplasma Symbiont of the Pea Aphid, Acyrthosiphon pisum (Insecta: Homoptera)

From a laboratory strain of the pea aphid, Acyrthosiphon pisum, we discovered a previously unknown facultative endosymbiotic bacterium. Molecular phylogenetic analysis based on 16S ribosomal DNA revealed that the bacterium is a member of the genus Spiroplasma. The Spiroplasma organism showed stable vertical transmission through successive generations of the host. Injection of hemolymph from infected insects into uninfected insects established a stable infection in the recipients. The Spiroplasma symbiont exhibited negative effects on growth, reproduction, and longevity of the host, particularly in older adults. Of 58 clonal strains of A. pisum established from natural populations in central Japan, 4 strains possessed the Spiroplasma organism.     

Thiomicrospira pelophila,gen. n., sp. n., a new obligately chemolithotrophic colourless sulfur bacterium

From marine mud flats a very thin, comma- or spiral-shaped bacterium was isolated. The new organism was an obligately chemolithotrophic sulfur bacterium. Its physiology was found to be essentially similar to that ofThiobacillus thioparus. Because of the spirillum-like appearance it was proposed to classify this bacterium into a new genusThiomicrospira, with the species nameTms. pelophila. Tms. pelophila and a marineT. thioparus, which was isolated from the same mud, occupy different niches in this habitat.Tms. pelophila has a remarkable sulfide-tolerance as compared withT. thioparus. This property could be used for the specific enrichment ofTms. pelophila. The organism was also readily isolated in pure culture by filtering mud suspensions through a 0.22 Μm membrane filter.     

Spore Dormancy Mutants of Phycomyces

Rivero, F., and Cerdá-Olmedo, E. 1994. Spore dormancy mutants of Phycomyces. Experimental Mycology 18, 221-229. The spores of the Zygomycete Phycomyces blakesleeanus are called dormant because few of them germinate when placed in a medium that sustains mycelial growth and development. Nearly all the spores germinate after activation, that is, exposure to heat or certain chemicals. We have looked for mutants whose spores would not need activation. Nine mutants formed authentic, but transient spores, which germinated spontaneously in the sporangium. Mutant mycelia had lower alcohol and aldehyde dehydrogenase activities and less glycogen than wild-type mycelia. The spontaneous germination and the metabolic alterations are attributed to the same recessive mutations. No differences were found between mutants and wild type in the cyclic AMP and fructose 2,6-bisphosphate concentrations in immature sporangia and the trehalase activity in the mycelia. In another mutant the spore primordia did not form spores, but remained viable for some time in the sporangium. The mutants were difficult to keep in the laboratory (except as lyophils); this stresses the importance of preventing spore germination in the sporangium.     

Ultrastructure of the nematode pathogen, Bacillus penetrans

The ultrastructure and development of Bacillus penetrans in root-knot nematodes, Meloidogyne spp., was studied with a transmission electron microscope. Host infection was by a germ tube from the cup-shaped sporangium containing the endospore. The prokaryotic vegetative cells contained septa formed by an ingrowth of the inner layer of the trilaminate cell wall and were associated with mesosomes. Structure of the endospore was similar to other bacteria with a spore protoplast enclosed within two cortical layers and three spore coats. An exosporium which may function in attachment and host specificity surrounded the endospore. Ultrastructural changes accompanying sporulation were similar to those reported for other endospore-forming bacteria but with some parasite specialization. The filamentous vegetative growth was characteristic of some Actinomycetales. Endospore development at the apices of dichotomously branched filaments of the thallus resembled the genus Actinobifida.     

A new anaerobic bacterium,forming up to five endospores per cell —Anaerobacter polyendosporus gen. et spec. nov.

An obligately anaerobic sporeforming bacterium assigned to a new genus and species Anaerobacter polyendosporus gen. et spec. nov. is described. Characteristic features distinguishing the bacterium from known anaerobic sporeformers were variable cell shape, including spherical, the ability to form up to five endospores per cell, diffusive distribution of reserve polysaccharide throughout the cytoplasm, independence from growth factors. The eubacterial nature of the organism was revealed by its sensitivity to 1 mg/l of streptomycin, rifampicin, penicillin and to lysozyme. It belonged to Firmicutes by the type of cell wall structure. The cell wall consisted of one layer; the outer membrane was absent. The cells were not motile. The spores were spherical or oval, heat-resistant, contained dipicolinic acid and had typical endospore structure. Cortex, coats, spore coare, and in most cases exosporium could be distinguished. The bacterium fermented carbohydrates, but not amino acids. The products of fermentation included ethanol, acetate, lactate, butyrate, butanol, H₂ and CO₂. Sulfate or nitrate could not be used as electron acceptors, but nitrite was reduced to NH ₄ ⁺ in a dissimilatory process. The bacterium was capable of fixing N₂. The G + C content of the DNA was 29 mol %. The bacterium was isolated from meadow-gley soil.     

Structural and physiological changes during sporangial development inAchlya intricata beneke

Summary Sporangial development in the zoosporic fungusAchlya intricata has been studied using light microscopy, a plasmolytic technique, KCl-filled microelectrodes and ion-selective microelectrodes. The completion of cleavage (spore delimitation) is accompanied by a change in appearance of the sporangium, loss of turgor and membrane potential, decrease in volume and release of K⁺. The measured loss of K⁺ is in agreement with previous measurements of extracellular ionic currents around developing sporangia ofAchlya using a vibrating probe. The relationship between these changes and the mechanism of spore liberation is discussed.     

Chemical and Morphological Studies of Bacterial Spore Formation : I. The Formation of Spores in Bacillus cereus

Experimental conditions were developed whereby a culture of Bacillus cereus formed spores with reasonable synchrony following a growth cycle of some 8 hours. The cytology of this metamorphosis was studied by dark phase contrast, bright-field microscopy and electron microscopy of thin sections. Particular attention has been paid to the changes in chromatin patterns and these have been correlated with quantitative chemical estimations of the nucleic acids. The cell commencing sporulation contains two compact chromatin bodies and twice the spore amount of deoxyribonucleic acid. Following fusion of the two chromatin bodies, one-half of this chromatin becomes located at a cell end. A transverse septum growing inwards from, and remaining attached to, the inner surface of the cell wall separates this end-piece of chromatin and some associated cytoplasm from the rest of the cell to form the primordial spore. Although the synthesis of deoxyribonucleic acid ceases during the segregation process, it recommences in this organism and continues at a linear rate as the spore develops. Tracer studies with radioactive phosphorus indicated that this further synthesis is confined to the non-spore portion of the sporangium. Although the net synthesis of ribonucleic acid ceased prior to the onset of sporogenesis, some evidence of a turnover of this fraction during the sporulation process was found.     

The epiphytic diatom flora of mosses from Strømness Bay area, South Georgia

A total of 104 taxa, belonging to 27 genera, were recorded from 36 moss samples collected in the Strømness Bay area. Cluster analysis revealed two groups of samples. Group 1 consisted of the Achnanthes incognita–Fragilaria germainii assemblage and the Eunotia paludosa var. paludosa assemblage. Group 2 included the Pinnularia borealis assemblage. The major ecological factor dividing the two groups appears to be the moisture content of the samples. Group 1 represents wetter conditions, and the second group is found on very dry mosses. The assemblages of group 1 were separated from each other by pH. The Achnanthes incognita–Fragilaria germainii assemblage occurs in wet mosses with a circumneutral pH. The Eunotia paludosa var. paludosa assemblage prefers bryophyte habitats with low pH values. The size of Pinnularia borealis drops significantly according to the dryness of the moss habitat.     

A new opiine (Hymenoptera,Braconidae) from Australia with discussion of Diachasma Foerster

A new species of Opiinae, Diachasma dentatum Shirley, Restuccia & Ly, is described from Australia. This species is similar to several other Australian opiines previously described or included in the genus Diachasma, but the mandibles are unusually broad, nearly exodont. Notable differences between Australian and Palaearctic Diachasma are discussed. Diachasma tasmaniae Fischer, 1995, originally described from Tasmania and New South Wales, is newly recorded from Victoria. Diachasma rufipes Szépligeti, 1905 is transferred to Notiopambolus, new combination.     

A new species of <Emphasis Type="Italic">Vanilla</Emphasis> (Orchidaceae: Vanilloideae) from São Paulo,Brazil

A new orchid species, Vanilla paludosa, native to marshy forests close to cerrado vegetation in the state of São Paulo, Brazil, is described and illustrated. The relationship of V. paludosa to other species of the genus is discussed.     

Cloning and characterization of an ftsZ homologue from a bacterial symbiont of Drosophila melanogaster

A 1194 by open reading frame that codes for a 398 amino acid peptide was cloned from a λgt11 library of Drosophila melanogaster genomic DNA. The predicted peptide sequence is very similar to three previously characterized protein sequences that are encoded by the ftsZ genes in Escherichia coli, Bacillus subtilis and Rhizobium meliloti. The FtsZ protein has a major role in the initiation of cell division in prokaryotic cells. Using a tetracycline treatment that eradicates bacterial parasites from insects, the ftsZ homologue has been found to be derived from a bacterium that lives within the strain. However, polymerase chain reaction (PCR) amplification of the gene from treated embryos suggests that it is not derived from a gut bacterium. Nevertheless, by amplifying and characterizing part of the 16S rRNA from this bacterium we have been able to demonstrate that it is a member of the genus Wolbachia, a parasitic organism that infects, and disturbs the sexual cycle of various strains of Drosophila simulans. We suggest that this ftsZ homologue is implicated in the cell division of Wolbachia, an organism that fails to grow outside the host organism. Sequence and alignment analysis of this ftsZ homologue show the presence of a potential GTP-binding motif indicating that it may function as a GTPase. The consequences of this function particularly with respect to its role in cell division are discussed.     

A new genus of Early Devonian plants with novel strobilar structures and vegetative appendages from the Posongchong Formation of Yunnan,China

A new plant, Dibracophyton acrovatum gen. et sp. nov., is described from the Lower Devonian (Pragian) Posongchong Formation of Wenshan District, southeastern Yunnan, China. The plant has creeping axes from which arise vegetative and fertile axes. The vegetative axes helically bear lateral dichotomous appendages with curved or round tips. The fertile axes possess terminal strobili with numerous fertile units arranged in irregular helices. Each fertile unit consists of a stalked long-elliptical sporangium, with dehiscence into two equal valves, and two discrete long-ovate bracts covering sporangium from above–below directions. A new genus and species is thus established based on these characters and is temporarily regarded as incertae sedis of Tracheophyta, although it perhaps closes to the barinophytes in affinity. Detailed comparisons with other plants having a similar architecture, i.e., sporangia closely associated with modified vegetative structures, are made. The discovery of D. acrovatum further enriches the composition of the Posongchong flora and demonstrates great morphological disparity of the Early Devonian vascular plants.     

Binding of Cyt1Aa and Cry11Aa Toxins of Bacillus thuringiensis Serovar israelensis to Brush Border Membrane Vesicles of Tipula paludosa (Diptera: Nematocera) and Subsequent Pore Formation

Bacillus thuringiensis serovar israelensis (B. thuringiensis subsp. israelensis) produces four insecticidal crystal proteins (ICPs) (Cry4A, Cry4B, Cry11A, and Cyt1A). Toxicity of recombinant B. thuringiensis subsp. israelensis strains expressing only one of the toxins was determined with first instars of Tipula paludosa (Diptera: Nematocera). Cyt1A was the most toxic protein, whereas Cry4A, Cry4B, and Cry11A were virtually nontoxic. Synergistic effects were recorded when Cry4A and/or Cry4B was combined with Cyt1A but not with Cry11A. The binding and pore formation are key steps in the mode of action of B. thuringiensis subsp. israelensis ICPs. Binding and pore-forming activity of Cry11Aa, which is the most toxic protein against mosquitoes, and Cyt1Aa to brush border membrane vesicles (BBMVs) of T. paludosa were analyzed. Solubilization of Cry11Aa resulted in two fragments, with apparent molecular masses of 32 and 36 kDa. No binding of the 36-kDa fragment to T. paludosa BBMVs was detected, whereas the 32-kDa fragment bound to T. paludosa BBMVs. Only a partial reduction of binding of this fragment was observed in competition experiments, indicating a low specificity of the binding. In contrast to results for mosquitoes, the Cyt1Aa protein bound specifically to the BBMVs of T. paludosa, suggesting an insecticidal mechanism based on a receptor-mediated action, as described for Cry proteins. Cry11Aa and Cyt1Aa toxins were both able to produce pores in T. paludosa BBMVs. Protease treatment with trypsin and proteinase K, previously reported to activate Cry11Aa and Cyt1Aa toxins, respectively, had the opposite effect. A higher efficiency in pore formation was observed when Cyt1A was proteinase K treated, while the activity of trypsin-treated Cry11Aa was reduced. Results on binding and pore formation are consistent with results on ICP toxicity and synergistic effect with Cyt1Aa in T. paludosa.