A Robust and Universal Metaproteomics Workflow for Research Studies and Routine Diagnostics Within 24 h Using Phenol Extraction, FASP Digest, and the MetaProteomeAnalyzer

The investigation of microbial proteins by mass spectrometry (metaproteomics) is a key technology for simultaneously assessing the taxonomic composition and the functionality of microbial communities in medical, environmental, and biotechnological applications. We present an improved metaproteomics workflow using an updated sample preparation and a new version of the MetaProteomeAnalyzer software for data analysis. High resolution by multidimensional separation (GeLC, MudPIT) was sacrificed to aim at fast analysis of a broad range of different samples in less than 24 h. The improved workflow generated at least two times as many protein identifications than our previous workflow, and a drastic increase of taxonomic and functional annotations. Improvements of all aspects of the workflow, particularly the speed, are first steps toward potential routine clinical diagnostics (i.e., fecal samples) and analysis of technical and environmental samples. The MetaProteomeAnalyzer is provided to the scientific community as a central remote server solution at www.mpa.ovgu.de.Peer Reviewe

Cryptic photosynthesis, Extrasolar planetary oxygen without a surface biological signature

On the Earth, photosynthetic organisms are responsible for the production of virtually all of the oxygen in the atmosphere. On the land, vegetation reflects in the visible, leading to a red edge that developed about 450 Myr ago and has been proposed as a biosignature for life on extrasolar planets. However, in many regions of the Earth, and particularly where surface conditions are extreme, for example in hot and cold deserts, photosynthetic organisms can be driven into and under substrates where light is still sufficient for photosynthesis. These communities exhibit no detectable surface spectral signature to indicate life. The same is true of the assemblages of photosynthetic organisms at more than a few metres depth in water bodies. These communities are widespread and dominate local photosynthetic productivity. We review known cryptic photosynthetic communities and their productivity. We link geomicrobiology with observational astronomy by calculating the disk-averaged spectra of cryptic habitats and identifying detectable features on an exoplanet dominated by such a biota. The hypothetical cryptic photosynthesis worlds discussed here are Earth-analogs that show detectable atmospheric biomarkers like our own planet, but do not exhibit a discernable biological surface feature in the disc-averaged spectrum.Comment: 23 pages, 2 figures, Astrobiology (TBP) - updated Table 1, typo in detectable O2 correcte

Estimating Net Photosynthesis of Biological Soil Crusts in the Atacama Using Hyperspectral Remote Sensing

Biological soil crusts (BSC) encompassing green algae, cyanobacteria, lichens, bryophytes, heterotrophic bacteria and microfungi are keystone species in arid environments because of their role in nitrogen- and carbon-fixation, weathering and soil stabilization, all depending on the photosynthesis of the BSC. Despite their importance, little is known about the BSCs of the Atacama Desert, although especially crustose chlorolichens account for a large proportion of biomass in the arid coastal zone, where photosynthesis is mainly limited due to low water availability. Here, we present the first hyperspectral reflectance data for the most wide-spread BSC species of the southern Atacama Desert. Combining laboratory and field measurements, we establish transfer functions that allow us to estimate net photosynthesis rates for the most common BSC species. We found that spectral differences among species are high, and differences between the background soil and the BSC at inactive stages are low. Additionally, we found that the water absorption feature at 1420 nm is a more robust indicator for photosynthetic activity than the chlorophyll absorption bands. Therefore, we conclude that common vegetation indices must be taken with care to analyze the photosynthesis of BSC with multispectral data

Common loss of far-red light photoacclimation in cyanobacteria from hot and cold deserts: a case study in the Chroococcidiopsidales

Deserts represent an extreme challenge for photosynthetic life. Despite their aridity, they are often inhabited by diverse microscopic communities of cyanobacteria. These organisms are commonly found in lithic habitats, where they are partially sheltered from extremes of temperature and UV radiation. However, living under the rock surface imposes additional constraints, such as limited light availability, and enrichment of longer wavelengths than are typically usable for oxygenic photosynthesis. Some cyanobacteria from the genus Chroococcidiopsis can use this light to photosynthesize, in a process known as far-red light photoacclimation, or FaRLiP. This genus has commonly been reported from both hot and cold deserts. However, not all Chroococcidiopsis strains carry FaRLiP genes, thus motivating our study into the interplay between FaRLiP and extreme lithic environments. The abundance of sequence data and strains provided the necessary material for an in-depth phylogenetic study, involving spectroscopy, microscopy, and determination of pigment composition, as well as gene and genome analyses. Pigment analyses revealed the presence of red-shifted chlorophylls d and f in all FaRLiP strains tested. In addition, eight genus-level taxa were defined within the encompassing Chroococcidiopsidales, clarifying the phylogeny of this long-standing polyphyletic order. FaRLiP is near universally present in a generalist genus identified in a wide variety of environments, Chroococcidiopsis sensu stricto, while it is rare or absent in closely related, extremophile taxa, including those preferentially inhabiting deserts. This likely reflects the evolutionary process of gene loss in specialist lineages

Common loss of far-red light photoacclimation in cyanobacteria from hot and cold deserts: a case study in the Chroococcidiopsidales

Deserts represent an extreme challenge for photosynthetic life. Despite their aridity, they are often inhabited by diverse microscopic communities of cyanobacteria. These organisms are commonly found in lithic habitats, where they are partially sheltered from extremes of temperature and UV radiation. However, living under the rock surface imposes additional constraints, such as limited light availability, and enrichment of longer wavelengths than are typically usable for oxygenic photosynthesis. Some cyanobacteria from the genus Chroococcidiopsis can use this light to photosynthesize, in a process known as far-red light photoacclimation, or FaRLiP. This genus has commonly been reported from both hot and cold deserts. However, not all Chroococcidiopsis strains carry FaRLiP genes, thus motivating our study into the interplay between FaRLiP and extreme lithic environments. The abundance of sequence data and strains provided the necessary material for an in-depth phylogenetic study, involving spectroscopy, microscopy, and determination of pigment composition, as well as gene and genome analyses. Pigment analyses revealed the presence of red-shifted chlorophylls d and f in all FaRLiP strains tested. In addition, eight genus-level taxa were defined within the encompassing Chroococcidiopsidales, clarifying the phylogeny of this long-standing polyphyletic order. FaRLiP is near universally present in a generalist genus identified in a wide variety of environments, Chroococcidiopsis sensu stricto, while it is rare or absent in closely related, extremophile taxa, including those preferentially inhabiting deserts. This likely reflects the evolutionary process of gene loss in specialist lineages

The green algal underground : evolutionary secrets of desert cells

Author Posting. © American Institute of Biological Sciences, 2008. This article is posted here by permission of American Institute of Biological Sciences for personal use, not for redistribution. The definitive version was published in BioScience 58 (2008): 114-122, doi:10.1641/B580206.Microscopic, unicellular, free-living green algae are found in desert microbiotic crusts worldwide. Although morphologically simple, green algae in desert crusts have recently been found to be extraordinarily diverse, with membership spanning five green algal classes and encompassing many taxa new to science. This overview explores this remarkable diversity and its potential to lead to new perspectives on the diversity and evolution of green plants. Molecular systematic and physiological data gathered from desert taxa demonstrate that these algae are long-term members of desert communities, not transient visitors from aquatic habitats. Variations in desiccation tolerance and photophysiology among these algae include diverse evolutionary innovations that developed under selective pressures in the desert. Combined with the single embryophyte lineage to which more familiar terrestrial green plants belong, multiple desert green algal lineages provide independent evolutionary units that may enhance understanding of the evolution and ecology of eukaryotic photosynthetic life on land.This work was supported by grants from the National Aeronautics and Space Administration, Exobiology Program (EXB02-0042-0054) to L. A. L. and Z. G. C., from the National Science Foundation (DEB- 0529737) to L. A. L., and from the University of Connecticut Research Foundation to Z. G. C

Biological soil crusts of Arctic Svalbard and of Livingston Island, Antarctica

Biological soil crusts (BSCs) occur in arid and semi-arid regions worldwide including the Polar Regions. They are important ecosystem engineers, and their composition and areal coverage should be understood before assessing key current functional questions such as their role in biogeochemical nutrient cycles and possible climate change scenarios. Our aim was to investigate the variability of BSCs from Arctic Svalbard and the Antarctic Island, Livingston, using vegetation surveys based on classification by functional group. An additional aim was to describe the structure of BSCs and represent a classification system that can be used in future studies to provide a fast and efficient way to define vegetation type and areal coverage. Firstly, this study demonstrates huge areas occupied by BSCs in Arctic Svalbard, with up to 90 % of soil surface covered, dominated by bryophytes and cyanobacteria, and showing an unexpectedly high variability in many areas. Livingston Island has lower percentage coverage, up to 55 %, but is dominated by lichens. Our findings show that both Polar Regions have varied BSC coverage, within the sites and between them, especially considering their harsh climates and latitudinal positions. Secondly, we have classified the BSCs of both areas into a system that describes the dominant functional groups and local geography, creating a simple scheme that allows easy identification of the prevailing vegetation type. Our results represent the first contribution to the description of BSCs based on their functional group composition in Polar Regions

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