Cultivation and complete genome sequencing of Gloeobacter kilaueensis sp. nov., from a lava cave in Kilauea Caldera, Hawai'i.

The ancestor of Gloeobacter violaceus PCC 7421(T) is believed to have diverged from that of all known cyanobacteria before the evolution of thylakoid membranes and plant plastids. The long and largely independent evolutionary history of G. violaceus presents an organism retaining ancestral features of early oxygenic photoautotrophs, and in whom cyanobacteria evolution can be investigated. No other Gloeobacter species has been described since the genus was established in 1974 (Rippka et al., Arch Microbiol 100:435). Gloeobacter affiliated ribosomal gene sequences have been reported in environmental DNA libraries, but only the type strain's genome has been sequenced. However, we report here the cultivation of a new Gloeobacter species, G. kilaueensis JS1(T), from an epilithic biofilm in a lava cave in Kilauea Caldera, Hawai'i. The strain's genome was sequenced from an enriched culture resembling a low-complexity metagenomic sample, using 9 kb paired-end 454 pyrosequences and 400 bp paired-end Illumina reads. The JS1(T) and G. violaceus PCC 7421(T) genomes have little gene synteny despite sharing 2842 orthologous genes; comparing the genomes shows they do not belong to the same species. Our results support establishing a new species to accommodate JS1(T), for which we propose the name Gloeobacter kilaueensis sp. nov. Strain JS1(T) has been deposited in the American Type Culture Collection (BAA-2537), the Scottish Marine Institute's Culture Collection of Algae and Protozoa (CCAP 1431/1), and the Belgian Coordinated Collections of Microorganisms (ULC0316). The G. kilaueensis holotype has been deposited in the Algal Collection of the US National Herbarium (US# 217948). The JS1(T) genome sequence has been deposited in GenBank under accession number CP003587. The G+C content of the genome is 60.54 mol%. The complete genome sequence of G. kilaueensis JS1(T) may further understanding of cyanobacteria evolution, and the shift from anoxygenic to oxygenic photosynthesis.

Novel methylotrophic isolates from lake sediment, description of Methylotenera versatilis sp. nov. and emended description of the genus Methylotenera.

Phylogenetic positions, and genotypic and phenotypic characteristics of three novel methylotrophic isolates, strains 301(T), 30S and SIP3-4, from sediment of Lake Washington, Seattle, USA, are described. The strains were restricted facultative methylotrophs capable of growth on single carbon compounds (methylamine and methanol) in addition to a limited range of multicarbon compounds. All strains used the N-methylglutamate pathway for methylamine oxidation. Strain SIP3-4 possessed the canonical (MxaFI) methanol dehydrogenase, but strains 301(T) and 30S did not. All three strains used the ribulose monophosphate pathway for C1 assimilation. The major fatty acids in the three strains were C(16:0) and C(16:1)ω7c. The DNA G+C contents of strains 301(T) and SIP3-4 were 42.6 and 54.6 mol%, respectively. Based on 16S rRNA gene sequence phylogeny and the relevant phenotypic characteristics, strain SIP3-4 was assigned to the previously defined species Methylovorus glucosotrophus. Strains 301(T) and 30S were closely related to each other (100% 16S rRNA gene sequence similarity) and shared 96.6% 16S rRNA gene sequence similarity with a previously described isolate, Methylotenera mobilis JLW8(T). Based on significant genomic and phenotypic divergence with the latter, strains 301(T) and 30S represent a novel species within the genus Methylotenera, for which the name Methylotenera versatilis sp. nov. is proposed; the type strain is 301(T) (=VKM B-2679(T)=JCM 17579(T)). An emended description of the genus Methylotenera is provided.

Microbial community in acidic hydrothermal waters of volcanically active White Island, New Zealand.

We report the first description of the microbial community in a stream of acidic hydrothermal waters on volcanically active White Island, New Zealand, using both molecular and microbiological methods. alpha- and beta-Proteobacteria, green-sulfur bacteria, and uncultured Firmicutes were identified from the community DNA-based 16s rRNA gene library. The same bacterial groups and the Rhodophyte Cyanidium caldarium were represented in enrichment cultures. C. caldarium, two Firmicutes and an acidophilic alpha-Proteobacterium, Acidiphilium cryptum, were brought into pure culture. Bacteria cultured from the stream grow at pH > or =2, and the Cyanidium grows at pH 0.2.

Mechanistic distinctions between agrin and laminin-1 induced aggregation of acetylcholine receptors.

BACKGROUND: One of the earliest steps in synaptogenesis at the neuromuscular junction is the aggregation of nicotinic acetylcholine receptors at the postsynaptic membrane. This study presents quantitative analyses of receptor and alpha-Dystroglycan aggregation in response to agrin and laminin-1, alone or in combination. RESULTS: Both laminin and agrin increased overall expression of receptors on the plasma membrane. Following a 24 hour exposure, agrin increased the number of receptor aggregates but did not affect the number of alpha-Dystroglycan aggregates, while the reverse was true of laminin-1. Laminin also increased receptor concentration within aggregates, while agrin had no such effect. Finally, the spatial distribution of aggregates was indistinguishable from random in the case of laminin, while agrin induced aggregates were closer together than predicted by a random model. CONCLUSIONS: Agrin and laminin-1 both increase acetylcholine receptor aggregate size after 24 hours, but several lines of evidence indicate that this is achieved via different mechanisms. Agrin and laminin had different effects on the number and density of receptor and alpha-Dystroglycan aggregates. Moreover the random distribution of laminin induced (as opposed to agrin induced) receptor aggregates suggests that the former may influence aggregate size by simple mass action effects due to increased receptor expression.

Ultrastructure of a novel eurytele nematocyst of Carybdea alata Reynaud (Cubozoa, Cnidaria).

The ultrastructural characteristics of nematocysts from the cubozoan Carybdea alata Reynaud, 1830 (Hawaiian box jellyfish) were examined using light, scanning and transmission electron microscopy. We reclassified the predominant nematocyst in C. alata tentacles as a heterotrichous microbasic eurytele, based on spine, tubule and capsule measurements. These nematocysts exhibited a prominent and singular stylet, herein referred to as the lancet. Discharged nematocysts from fixed tentacle preparations displayed the following structures: a smooth shaft base, lamellae, a hemicircumferential fissure demarking the proximal end of a stratified lancet, and a gradually tapering tubule densely covered with large triangularly shaped spines. The lancet remained partially adjoined to the shaft base in a hinge-like fashion in rapidly fixed, whole-tentacle preparations. In contrast, this structure was not observed in discharged nematocyst preparations which involved multiple transfer steps prior to fixation. Various approaches were designed to detect this structure in the absence of fixative. Detached lancets were located in proximity to discharged tubules in undisturbed coverslip preparations of fresh tentacles. In addition, examination of embedded nematocysts from fresh tentacles laid on polyacrylamide gels revealed still-attached lancets. To examine the function of this structure in prey capture, Artemia sp. laden tentacles were prepared for scanning electron microscopy. While carapace exteriors exhibited structures proximal to the lancet, i.e., the nematocyst capsule and shaft base, neither tubule nor lancet structures were visible. Taken together, the morphological data suggested a series of events involved in the discharge of a novel eurytele from C. alata.

The Sauromatum guttatum appendix as an osmophore: excretory pathways, composition of volatiles and attractiveness to insects.

This report combines chemical, electron microscopic and ecological studies on the volatiles liberated by the Sauromatum guttatum appendix on D-Day, the day of inflorescence-opening and heat-production. More than 100 compounds from at least nine different chemical classes (monoterpenes, sesquiterpenes, fatty acids, ketones, alcohols, aldehydes, indole, and phenolic and sulphur compounds) are liberated during the thermogenic activity. The volatiles were identified using gas chromatography-mass spectrometry. Electron microscopy provides additional evidence that the endoplasmic reticulum (ER) interacts with the plasma membrane, creating novel routes of excretion of the volatiles to the exterior of the cell. It seems that the fusion event creates channels from the interior to the exterior of the cell. Furthermore, a multitubular body, conceivably originating in the ER, seems to fuse with the plasma membrane and to appear only on D-day. This multitubular body is closely associated with lipid bodies during heat-production and might be involved in the oxidation of lipids to volatile products. The foul odour produced by the appendix attracts at least 30 species of insects.