Molecular and morphological characterization of free-floating filamentous cyanobacterial mats from geothermal springs in the Philippines

A novel cyanobacterial mat type is characterized from near-neutral pH, low sulphide geothermal springs of 45-60 °C in the Philippines. Mats were free floating, several metres in diameter and several cm in thickness. The upper surface of mats was covered in a waxy scytonemin-like layer, solvent extracts of which absorbed light strongly at 384nm. Light microscopy revealed mats to posses highly ordered layers of air spaces at both the macroscopic and microscopic level, apparently as an adaptation to buoyancy. Morphospecies composition was exclusively filamentous, with Fischerella-like and Oscillatoria-like taxa closely associated throughout mats. Abundant heterocystous cells were observed in Fischerella filaments, suggesting nitrogen fixation occurs in these mats. Morphological structure did not vary among mats from pools of different temperature, but several 16S rDNA-defined genotypes were resolved by DGGE with some displaying greater thermophily than others. Sequencing of fourteen DGGE bands (Genbank accession numbers: AY236467-AY236480) yielded nine novel Fischerella sequences, whilst the five Oscillatoria sequences showed high similarity to other thermophilic Oscillatoria sequences. These data are relevant to astrobiology in that they expand our knowledge of oxygenic photosynthetic community diversity in geothermal environments, which serve as modern analogues for early life on Earth and other planets. Acknowledgements The authors are extremely grateful to the Philippine Institute of Volcanology and Seismology (PHIVOLCS) for advice and assistance with fieldwork. This work was supported by grants awarded by The University of Hong Kong CRCG Seed Funding for Basic Research and Small Projects programmes.published_or_final_versio

Optimization of laccase production by Pycnoporus sanguineus in submerged liquid culture

The white-rot fungus Pycnoporus sanguineus produces laccase under a range of C/N ratios in submerged liquid culture. Enzyme production was increased 50 fold in the presence of 20 μM xylidine to a maximum of 1368 U L-1 in a high carbon low nitrogen medium. Slight repression of enzyme production was observed in high nitrogen culture medium. Other potential inducers were less effective (Tween 80, wood fibres) and reduced the stimulation observed by xylidine alone when included in the same culture medium. Veratryl alcohol failed to stimulate laccase production. Activity of the enzyme activity in crude culture nitrate was stable at temperatures of 35 C and below with a pH optimum of 3.0. The laccase of P. sanguineus was identified as a ca 65 kDa protein produced as multiple isoforms.published_or_final_versio

Identification and characterization of thermophilic Synechococcus spp. isolates from Asian geothermal springs

Two thermophilic cyanobacterial strains, Ts and Bs, collected from Asian geothermal springs were identified morphologically and phylogenetically as Synechococcus in the order Chroococcales and were isolated into axenic cultures. In addition to the high similarities between their full 16S rRNA gene sequences, both strains also shared similar pigment profiles and fatty acid compositions but with varied ratios. Strain Ts had elevated levels of photoprotective pigments such as carotenoid and scytonemin even after prolonged culture under identical laboratory conditions, whereas strain Bs produced more chlorophyll a per unit cell volume, perhaps resulting from UV adaptation in the natural habitats. In addition, strain Ts had more content than strain Bs in terms of the total fatty acids and the proportion of unsaturated fatty acids. Neither isolate was able to fix nitrogen, and they had zero susceptibility to ampicillin and streptomycin. © 2007 NRC.published_or_final_versio

Population structure in a Philippines hot spring microbial mat

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Cyanobacteria and chloroflexi-dominated hypolithic colonization of quartz at the hyper-arid core of the Atacama Desert, Chile

Quartz stones are ubiquitous in deserts and are a substrate for hypoliths, microbial colonists of the underside of such stones. These hypoliths thrive where extreme temperature and moisture stress limit the occurrence of higher plant and animal life. Several studies have reported the occurrence of green hypolithic colonization dominated by cyanobacteria. Here, we describe a novel red hypolithic colonization from Yungay, at the hyper-arid core of the Atacama Desert in Chile. Comparative analysis of green and red hypoliths from this site revealed markedly different microbial community structure as revealed by 16S rRNA gene clone libraries. Green hypoliths were dominated by cyanobacteria (Chroococcidiopsis and Nostocales phylotypes), whilst the red hypolith was dominated by a taxonomically diverse group of chloroflexi. Heterotrophic phylotypes common to all hypoliths were affiliated largely to desiccation-tolerant taxa within the Actinobacteria and Deinococci. Alphaproteobacterial phylotypes that affiliated with nitrogen-fixing taxa were unique to green hypoliths, whilst Gemmatimonadetes phylotypes occurred only on red hypolithon. Other heterotrophic phyla recovered with very low frequency were assumed to represent functionally relatively unimportant taxa. © 2010 The Author(s).published_or_final_versionSpringer Open Choice, 21 Feb 201

Low-diversity fungal assemblage in an Antarctic Dry Valleys soil

The McMurdo Dry Valleys of Antarctica present extreme environmental challenges. Life is restricted to patchy occurrence of lichens, mosses and invertebrates, plus soil microbial communities. Fungi have been described in lichen symbioses but relatively little is known about the occurrence of free-living soil fungi in the Dry Valleys. A challenge in estimating fungal species richness has been the extent to which estimates based on either cultivation or environmental DNA reflect the active assemblage in cold-arid soils. Here, we describe analysis for inland Dry Valleys soil of environmental DNA and RNA (cDNA) to infer total and putative metabolically active assemblages, respectively, plus cultivation approaches using a variety of laboratory growth conditions. Environmental sequences indicated a highly restricted assemblage of just seven phylotypes that affiliated phylogenetically within two known genera, Helicodendron and Zalerion, plus previously unidentified fungal phylotypes. None of the commonly encountered molds and mitosporic genera recorded from maritime Antarctic locations were encountered. A striking difference was observed in the frequency of recovery for phylotypes between libraries. This suggests that both species richness and beta diversity estimates based on DNA libraries have the potential to misinform putatively active assemblages. Cultivation yielded a cold-tolerant Zalerion strain that affiliated with DNA and RNA library clones, and a psychrotrophic yeast (Debaryomyces hansenii), which was not detected using either culture-independent approach. © 2011 The Author(s).published_or_final_versionSpringer Open Choice, 21 Feb 201

Community structure of free-floating filamentous cyanobacterial mats from the Wonder Lake geothermal springs in the Philippines

Cyanobacterial mats were characterized from pools of 45-60 °C in near-neutral pH, low-sulphide geothermal springs in the Philippines. Mat structure did not vary with temperature. All mats possessed highly ordered layers of airspaces at both the macroscopic and microscopic level, and these appear to be an adaptation to a free-floating growth habit. Upper mat layers supported biomass with elevated carotenoid:chlorophyll a ratios and an as yet uncharacterized waxy layer on the dorsal surface. Microscopic examination revealed mats comprised a single Fischerella morphotype, with abundant heterocysts throughout mats at all temperatures. Molecular analysis of mat community structure only partly matched morphological identification. All samples supported greater 16S rDNA-defined diversity than morphology suggested, with a progressive loss in the number of genotypes with increasing temperature. Fischerella-like sequences were recovered from mats occurring at all temperatures, but some mats also yielded Oscillatoria-like sequences, although corresponding phenotypes were not observed. Phylogenetic analysis revealed that Fischerella-like sequences were most closely affiliated with Fischerella major and the Oscillatoria-like sequences with Oscillatoria amphigranulata. © 2005 NRC Canada.published_or_final_versio

Hypolithic Microbial Community of Quartz Pavement in the High-Altitude Tundra of Central Tibet

The hypolithic microbial community associated with quartz pavement at a high-altitude tundra location in central Tibet is described. A small-scale ecological survey indicated that 36% of quartz rocks were colonized. Community profiling using terminal restriction fragment length polymorphism revealed no significant difference in community structure among a number of colonized rocks. Real-time quantitative PCR and phylogenetic analysis of environmental phylotypes obtained from clone libraries were used to elucidate community structure across all domains. The hypolithon was dominated by cyanobacterial phylotypes (73%) with relatively low frequencies of other bacterial phylotypes, largely represented by the chloroflexi, actinobacteria, and bacteriodetes. Unidentified crenarchaeal phylotypes accounted for 4% of recoverable phylotypes, while algae, fungi, and mosses were indicated by a small fraction of recoverable phylotypes

Hypolithic and soil microbial community assembly along an aridity gradient in the Namib Desert

The Namib Dessert is considered the oldest desert in the world and hyperarid for the last 5 million years. However, the environmental buffering provided by quartz and other translucent rocks supports extensive hypolithic microbial communities. In this study, open soil and hypolithic microbial communities have been investigated along an East–West transect characterized by an inverse fog-rainfall gradient. Multivariate analysis showed that structurally different microbial communities occur in soil and in hypolithic zones. Using variation partitioning, we found that hypolithic communities exhibited a fog-related distribution as indicated by the significant East– West clustering. Sodium content was also an important environmental factor affecting the composition of both soil and hypolithic microbial communities. Finally, although null models for patterns in microbial communities were not supported by experimental data, the amount of unexplained variation (68–97 %) suggests that stochastic processes also play a role in the assembly of such communities in the Namib Desert.Web of Scienc