Eukaryotic Microbial Communities Research Articles

Characterization of Prokaryotic and Eukaryotic Microbial Community in Pacific White Shrimp Ponds

This research profiles the community compositions and dominant taxonomies of prokaryotic and eukaryotic microbes in 30 samples of the water from 5 different Pacific white shrimp ponds. The V4 region of the 16S rRNA gene and 18S rRNA gene were sequenced by high-throughput sequencing technology. Total of 1,387,317 16S rRNA and 1,612,056 18S rRNA gene fragments were selected for classification, including 3,841 prokaryotic Operational Taxonomic Units (OTUs) and 990 eukaryotic OTUs. It’s observed that all of the 16S rRNA sequences were affiliated with at least 47 bacteria divisions and 18S rRNA sequences were affiliated with 50 eukaryotic divisions, respectively. Among all 30 samples, the dominant prokaryotic and eukaryotic community at phylum level shared considerable similarity in composition but not in abundance. The dominant prokaryotic community included Actinobacteria, Proteobacteria, Cyanobacteria, Planctomycetes, Verrucomicrobia, Bacteroidetes, Chlorobi, Chloroflexi, Firmicutes and Spirochaetes. Cercozoa, Chlororhyta, Arthropoda, Stramenopiles-unidentified, Fungi-unidentified, Prymnesiophyceae, Ciliophora, Mollusca, Choanomonada and Jakobida were the dominant compositions of the eukaryotic. Similarly, significant difference existed at the genus level among the 30 samples. Results of richness and diversity showed that prokaryotic and eukaryotic microbes possessed complex community compositions in 5 ponds. While in different periods and different ponds, the value of Chao, Ace, Shannon and Simpson index were not significantly different (P >0.05).

Large variability of bathypelagic microbial eukaryotic communities across the world's oceans.

In this work, we study the diversity of bathypelagic microbial eukaryotes (0.8-20 μm) in the global ocean. Seawater samples from 3000 to 4000 m depth from 27 stations in the Atlantic, Pacific and Indian Oceans were analyzed by pyrosequencing the V4 region of the 18S ribosomal DNA. The relative abundance of the most abundant operational taxonomic units agreed with the results of a parallel metagenomic analysis, suggesting limited PCR biases in the tag approach. Although rarefaction curves for single stations were seldom saturated, the global analysis of all sequences together suggested an adequate recovery of bathypelagic diversity. Community composition presented a large variability among samples, which was poorly explained by linear geographic distance. In fact, the similarity between communities was better explained by water mass composition (26% of the variability) and the ratio in cell abundance between prokaryotes and microbial eukaryotes (21%). Deep diversity appeared dominated by four taxonomic groups (Collodaria, Chrysophytes, Basidiomycota and MALV-II) appearing in different proportions in each sample. Novel diversity amounted to 1% of the pyrotags and was lower than expected. Our study represents an essential step in the investigation of bathypelagic microbial eukaryotes, indicating dominating taxonomic groups and suggesting idiosyncratic assemblages in distinct oceanic regions.

Characterization of Bacterial, Archaeal and Eukaryote Symbionts from Antarctic Sponges Reveals a High Diversity at a Three-Domain Level and a Particular Signature for This Ecosystem.

Sponge-associated microbial communities include members from the three domains of life. In the case of bacteria, they are diverse, host specific and different from the surrounding seawater. However, little is known about the diversity and specificity of Eukarya and Archaea living in association with marine sponges. This knowledge gap is even greater regarding sponges from regions other than temperate and tropical environments. In Antarctica, marine sponges are abundant and important members of the benthos, structuring the Antarctic marine ecosystem. In this study, we used high throughput ribosomal gene sequencing to investigate the three-domain diversity and community composition from eight different Antarctic sponges. Taxonomic identification reveals that they belong to families Acarnidae, Chalinidae, Hymedesmiidae, Hymeniacidonidae, Leucettidae, Microcionidae, and Myxillidae. Our study indicates that there are different diversity and similarity patterns between bacterial/archaeal and eukaryote microbial symbionts from these Antarctic marine sponges, indicating inherent differences in how organisms from different domains establish symbiotic relationships. In general, when considering diversity indices and number of phyla detected, sponge-associated communities are more diverse than the planktonic communities. We conclude that three-domain microbial communities from Antarctic sponges are different from surrounding planktonic communities, expanding previous observations for Bacteria and including the Antarctic environment. Furthermore, we reveal differences in the composition of the sponge associated bacterial assemblages between Antarctic and tropical-temperate environments and the presence of a highly complex microbial eukaryote community, suggesting a particular signature for Antarctic sponges, different to that reported from other ecosystems.

Eukaryotic Microbial Communities Associated with Rock- dwelling Foliose Lichens: A Functional Morphological and Microecological Analysis

Lichens are widely recognized as important examples of a fungal-algal or fungal-cyanophyte symbiosis; and in some cases they are a major food source for some animal grazers such as caribou ( Rangifer tarandus ), especially in the Arctic during winter. However, relatively little is known about the ecology of their co-associated bacterial and protistan communities. This is one of the first reports of an analysis of microbial communities associated with rock-dwelling foliose lichens ( Flavoparmelia sp.), including a more detailed analysis of the microbial communities associated with segments of the shield-like, radially arranged lobes. Samples were taken from lichens on granite boulders beneath an oak and maple tree stand on the Lamont-Doherty Earth Observatory Campus, Palisades, N.Y. The bacteria and protist members of the lichen associated microbial communities are comparable to recently reported associations for foliose lichens growing on tree bark at the same locale, including the presence of large myxomycete plasmodial amebas, heterotrophic nanoflagellates, and naked and testate amebas. To obtain evidence of possible differences in the microecology of different portions of each radial lobe, three segments of the radial lobe in the shield-like lichen were sampled: 1) inner, more mature, central segment; 2) middle section linking the central and peripheral segments; and 3) outer, peripheral, usually broader, less closely attached segment. The mean densities (number/g) and biomasses (µg/g) of bacteria and heterotrophic nanoflagellates were highest in the older central segment and lowest in the peripheral segment of the radial lobes, especially when expressed on moist weight basis. Large myxomycete plasmodial amebas were typically located in the outermost segment of the radial lobe. The proportion of vannellid amebas ( Vannella spp. and Cochliopodium spp.) were significantly more abundant in the samples of the inner lobes compared to non-vannellid amebas that were more prevalent in the outer lobes. The outer segment of the thallus lobe was typically more spongiose and absorbed more water per unit weight (based on a wet/dry-weight ratio) than the innermost segment. In general, patterns of densities and taxonomic composition of bacteria and eukaryotic microbes intergraded from the inner most segment to the outer part of each lobe – indicating a possible microecological gradient, coincident with the age-related and morphological radial gradations of the lobe. Overall, the evidence shows that the radial variation in the morphology and age-related variables of the three lobe segments may affect the microenvironment of the lobe segments and hence influence the organization of the microbial communities within each segment.

A retrospective metagenomics approach to studying Blastocystis.

Blastocystis is a common single-celled intestinal parasitic genus, comprising several subtypes. Here, we screened data obtained by metagenomic analysis of faecal DNA for Blastocystis by searching for subtype-specific genes in coabundance gene groups, which are groups of genes that covary across a selection of 316 human faecal samples, hence representing genes originating from a single subtype. The 316 faecal samples were from 236 healthy individuals, 13 patients with Crohn's disease (CD) and 67 patients with ulcerative colitis (UC). The prevalence of Blastocystis was 20.3% in the healthy individuals and 14.9% in patients with UC. Meanwhile, Blastocystis was absent in patients with CD. Individuals with intestinal microbiota dominated by Bacteroides were much less prone to having Blastocystis-positive stool (Matthew's correlation coefficient = -0.25, P < 0.0001) than individuals with Ruminococcus- and Prevotella-driven enterotypes. This is the first study to investigate the relationship between Blastocystis and communities of gut bacteria using a metagenomics approach. The study serves as an example of how it is possible to retrospectively investigate microbial eukaryotic communities in the gut using metagenomic datasets targeting the bacterial component of the intestinal microbiome and the interplay between these microbial communities.

A Multi Size-Level Assessment of Benthic Marine Communities in a Coastal Environment: Are They Different Sides of the Same Coin?

Organism body size has been demonstrated to be a discriminating element in shaping the response of living beings to environmental factors, thus playing a fundamental role in community structuring. Despite the importance of studies elucidating relations among communities of different size levels in ecosystems, the attempts that have been made in this sense are still very scarce and a reliable approach for these research still has to be defined. We characterized the benthic communities of bacteria, microbial eukaryotes, meiofauna and macrofauna in a coastal environment, encompassing a 10000-fold gradient in body size, testing and discussing a mixed approach of molecular fingerprinting for microbes and morphological observations for meio- and macrofauna. We found no correlation among structures of the different size-level communities: this suggests that community composition at one size-level could have no (or very low) influence on the community composition at other size-levels. Moreover, each community responds in a different way to the environmental parameters and with a degree of sensitivity which seems to increase with organism size. Therefore, our data indicate that the characterization of all the different size levels is clearly a necessity in order to study the dynamics really acting in a system.

Can dispersions be used for discriminating water quality status in coastal ecosystems? A case study on biofilm-dwelling microbial eukaryotes

Multivariate dispersions are an effective ecological indicator to evaluate the environmental heterogeneity and human disturbance in global ecological research. To explore the feasibility of the dispersions for assessing water quality status, a dataset of the biofilm-dwelling microbial eukaryote communities was studied in coastal waters of the Yellow Sea, northern China at two numerical resolutions. The community dispersion patterns represented a significant variability in both community structure and species composition in response to environmental heterogeneity. Multivariate approaches revealed that the species composition was the primary contributor to the community dispersion. Mantel analysis showed that the spatial variations in both community structure and species composition were significantly correlated with the changes of the nutrients, pH, dissolved oxygen (DO) and chemical oxygen demand (COD). The dispersion measures at both numerical resolutions were found to be remarkably related to the nutrient ammonium nitrogen and DO. Thus, we suggest that multivariate dispersions of the microbial eukaryote communities may be used as a potential bioindicator of water quality status in marine ecosystems.

Size-fractionated diversity of eukaryotic microbial communities in the Eastern Tropical North Pacific oxygen minimum zone

Oxygen minimum zones (OMZs) caused by water column stratification appear to expand in parts of the world's ocean, with consequences for marine biogeochemical cycles. OMZ formation is often fueled by high surface primary production, and sinking organic particles can be hotspots of interactions and activity within microbial communities. This study investigated the diversity of OMZ protist communities in two biomass size fractions (>30 and 30-1.6 μm filters) from the world's largest permanent OMZ in the Eastern Tropical North Pacific. Diversity was quantified via Illumina MiSeq sequencing of V4 region of 18S SSU rRNA genes in samples spanning oxygen gradients at two stations. Alveolata and Rhizaria dominated the two size fractions at both sites along the oxygen gradient. Community composition at finer taxonomic levels was partially shaped by oxygen concentration, as communities associated with versus anoxic waters shared only ∼32% of operational taxonomic unit (OTU) (97% sequence identity) composition. Overall, only 9.7% of total OTUs were recovered at both stations and under all oxygen conditions sampled, implying structuring of the eukaryotic community in this area. Size-fractionated communities exhibited different taxonomical features (e.g. Syndiniales Group I in the 1.6-30 μm fraction) that could be explained by the microniches created on the surface-originated sinking particles.

Quantifying the relative roles of selective and neutral processes in defining eukaryotic microbial communities.

We have a limited understanding of the relative contributions of different processes that regulate microbial communities, which are crucial components of both natural and agricultural ecosystems. The contributions of selective and neutral processes in defining community composition are often confounded in field studies because as one moves through space, environments also change. Managed ecosystems provide an excellent opportunity to control for this and evaluate the relative strength of these processes by minimising differences between comparable niches separated at different geographic scales. We use next-generation sequencing to characterize the variance in fungal communities inhabiting adjacent fruit, soil and bark in comparable vineyards across 1000 kms in New Zealand. By compartmentalizing community variation, we reveal that niche explains at least four times more community variance than geographic location. We go beyond merely demonstrating that different communities are found in both different niches and locations by quantifying the forces that define these patterns. Overall, selection unsurprisingly predominantly shapes these microbial communities, but we show the balance of neutral processes also have a significant role in defining community assemblage in eukaryotic microbes.

High diversity of protistan plankton communities in remote high mountain lakes in the European Alps and the Himalayan mountains.

We analyzed the genetic diversity (V4 region of the 18S rRNA) of planktonic microbial eukaryotes in four high mountain lakes including two remote biogeographic regions (the Himalayan mountains and the European Alps) and distinct habitat types (clear and glacier-fed turbid lakes). The recorded high genetic diversity in these lakes was far beyond of what is described from high mountain lake plankton. In total, we detected representatives from 66 families with the main taxon groups being Alveolata (55.0% OTUs97%, operational taxonomic units), Stramenopiles (34.0% OTUs97%), Cryptophyta (4.0% OTUs97%), Chloroplastida (3.6% OTUs97%) and Fungi (1.7% OTUs97%). Centrohelida, Choanomonada, Rhizaria, Katablepharidae and Telonema were represented by <1% OTUs97%. Himalayan lakes harbored a higher plankton diversity compared to the Alpine lakes (Shannon index). Community structures were significantly different between lake types and biogeographic regions (Fisher exact test, P < 0.01). Network analysis revealed that more families of the Chloroplastida (10 vs 5) and the Stramenopiles (14 vs 8) were found in the Himalayan lakes than in the Alpine lakes and none of the fungal families was shared between them. Biogeographic aspects as well as ecological factors such as water turbidity may structure the microbial eukaryote plankton communities in such remote lakes.

Analysis of Marine Fouling Microbial Communities Adhering to Carboxyl Modified MWCNTs-Filled PDMS Coating Surface during the Initial Stage of Biofouling

In this study, a polydimethylsiloxane (PDMS) coating filled with low concentrations of selected carboxyl modified multi-walled carbon nanotubes (cMWCNTs) has been fabricated. The antifouling properties of cMWCNTs-filled PDMS coatings were tested and the diversity level and succession phenomenon of marine fouling microbial communities were analyzed using single strand conformation polymorphism (SSCP) method. Marine adhesion test showed that cMWCNTs-filled PDMS coating presented decent antifouling property. SSCP analysis revealed that fouling prokaryotic species on the cMWCNTs-filled PDMS coating presented high and stable diversity level while diversity and dominance level of fouling eukaryotic species were relatively low, similar to those on PDMS alone and other four antifouling coating surface without nanoparticles, suggesting that the main mechanism by which cMWCNTs-mediated surface nanostructure improves antifouling capacity may have no direct relationship with the patterns in the context of succession dynamics of prokaryotic and eukaryotic microbial communities.

Contrasting elevational diversity patterns between eukaryotic soil microbes and plants

The diversity of eukaryotic macroorganisms such as animals and plants usually declines with increasing elevation and latitude. By contrast, the community structure of prokaryotes such as soil bacteria does not generally correlate with elevation or latitude, suggesting that differences in fundamental cell biology and/or body size strongly influence diversity patterns. To distinguish the influences of these two factors, soil eukaryotic microorganism community structure was investigated in six representative vegetation sites along an elevational gradient from forest to alpine tundra on Changbai Mountain in Northeast China, and compared with our previous determination of soil bacterial community structure along the same gradient. Using bar‐coded pyrosequencing, we found strong site differences in eukaryotic microbial community composition. However, diversity of the total eukaryotic microorganism community (or just the fungi or protists alone) did not correlate with elevation. Instead, the patterns of diversity and composition in the total eukaryotic microbial community (and in the protist community alone) were closely correlated with soil pH, suggesting that just as for bacteria, acidity is a particularly important determinant of eukaryotic microbial distributions. By contrast, as expected, plant diversity at the same sites declined along our elevational gradient. These results together suggest that elevational diversity patterns exhibited by eukaryotic microorganisms are fundamentally different from those of plants.

Protist community composition during early phytoplankton blooms in the naturally iron-fertilized Kerguelen area (Southern Ocean)

Abstract. Microbial eukaryotic community composition was examined by 18S rRNA gene tag pyrosequencing, during the early phase of spring phytoplankton blooms induced by natural iron fertilization, off Kerguelen Island in the Southern Ocean (KEOPS2 cruise). A total of 999 operational taxonomical units (OTUs), affiliated to 30 known high-level taxonomic groups, were retrieved from 16 samples collected in the upper 300 m water column. The alveolata group was the most abundant in terms of sequence number and diversity (696 OTUs). The majority of alveolata sequences were affiliated to Dinophyceae and to two major groups of marine alveolates (MALV-I and MALV-II). In the upper 180 m, only 13% of the OTUs were shared between of the fertilized stations and the reference site characterized by high-nutrient low-chlorophyll (HNLC) waters. Fungi and Cercozoa were present in iron-fertilized waters, but almost absent in the HNLC samples, while Haptophyta and Chlorophyta characterized the HNLC sample. Finally, the 300 m depth samples of all stations were differentiated by the presence of MALV-II and Radiolaria. Multivariate analysis, examining the level of similarity between different samples, showed that protistan assemblages differed significantly between the HNLC and iron-fertilized stations, but also between the diverse iron-fertilized blooms.

Oceanographic structure drives the assembly processes of microbial eukaryotic communities.

Arctic Ocean microbial eukaryote phytoplankton form subsurface chlorophyll maximum (SCM), where much of the annual summer production occurs. This SCM is particularly persistent in the Western Arctic Ocean, which is strongly salinity stratified. The recent loss of multiyear sea ice and increased particulate-rich river discharge in the Arctic Ocean results in a greater volume of fresher water that may displace nutrient-rich saltier waters to deeper depths and decrease light penetration in areas affected by river discharge. Here, we surveyed microbial eukaryotic assemblages in the surface waters, and within and below the SCM. In most samples, we detected the pronounced SCM that usually occurs at the interface of the upper mixed layer and Pacific Summer Water (PSW). Poorly developed SCM was seen under two conditions, one above PSW and associated with a downwelling eddy, and the second in a region influenced by the Mackenzie River plume. Four phylogenetically distinct communities were identified: surface, pronounced SCM, weak SCM and a deeper community just below the SCM. Distance–decay relationships and phylogenetic structure suggested distinct ecological processes operating within these communities. In the pronounced SCM, picophytoplanktons were prevalent and community assembly was attributed to water mass history. In contrast, environmental filtering impacted the composition of the weak SCM communities, where heterotrophic Picozoa were more numerous. These results imply that displacement of Pacific waters to greater depth and increased terrigenous input may act as a control on SCM development and result in lower net summer primary production with a more heterotroph dominated eukaryotic microbial community.

Solution hybrid selection capture for the recovery of functional full-length eukaryotic cDNAs from complex environmental samples.

Eukaryotic microbial communities play key functional roles in soil biology and potentially represent a rich source of natural products including biocatalysts. Culture-independent molecular methods are powerful tools to isolate functional genes from uncultured microorganisms. However, none of the methods used in environmental genomics allow for a rapid isolation of numerous functional genes from eukaryotic microbial communities. We developed an original adaptation of the solution hybrid selection (SHS) for an efficient recovery of functional complementary DNAs (cDNAs) synthesized from soil-extracted polyadenylated mRNAs. This protocol was tested on the Glycoside Hydrolase 11 gene family encoding endo-xylanases for which we designed 35 explorative 31-mers capture probes. SHS was implemented on four soil eukaryotic cDNA pools. After two successive rounds of capture, >90% of the resulting cDNAs were GH11 sequences, of which 70% (38 among 53 sequenced genes) were full length. Between 1.5 and 25% of the cloned captured sequences were expressed in Saccharomyces cerevisiae. Sequencing of polymerase chain reaction-amplified GH11 gene fragments from the captured sequences highlighted hundreds of phylogenetically diverse sequences that were not yet described, in public databases. This protocol offers the possibility of performing exhaustive exploration of eukaryotic gene families within microbial communities thriving in any type of environment.

Construction of sized eukaryotic cDNA libraries using low input of total environmental metatranscriptomic RNA.

BackgroundConstruction of high quality cDNA libraries from the usually low amounts of eukaryotic mRNA extracted from environmental samples is essential in functional metatranscriptomics for the selection of functional, full-length genes encoding proteins of interest. Many of the inserts in libraries constructed by standard methods are represented by truncated cDNAs due to premature stoppage of reverse transcriptase activity and preferential cloning of short cDNAs.ResultsWe report here a simple and cost effective technique for preparation of sized eukaryotic cDNA libraries from as low as three microgram of total soil RNA dominated by ribosomal and bacterial RNA. cDNAs synthesized by a template switching approach were size-fractionated by two dimensional agarose gel electrophoresis prior to PCR amplification and cloning. Effective size selection was demonstrated by PCR amplification of conserved gene families specific of each size class. Libraries of more than one million independent inserts whose sizes ranged between one and four kb were thus produced. Up to 80% of the insert sequences were homologous to eukaryotic gene sequences present in public databases.ConclusionsA simple and cost effective technique has been developed to construct sized eukaryotic cDNA libraries from environmental samples. This technique will facilitate expression cloning of environmental eukaryotic genes and contribute to a better understanding of basic biological and/or ecological processes carried out by eukaryotic microbial communities.

Effects of long-term differential fertilization on eukaryotic microbial communities in an arable soil: a multiple barcoding approach.

To understand the fine-scale effects of changes in nutrient availability on eukaryotic soil microorganisms communities, a multiple barcoding approach was used to analyse soil samples from four different treatments in a long-term fertilization experiment. We performed PCR amplification on soil DNA with primer pairs specifically targeting the 18S rRNA genes of all eukaryotes and three protist groups (Cercozoa, Chrysophyceae-Synurophyceae and Kinetoplastida) as well as the ITS gene of fungi and the 23S plastid rRNA gene of photoautotrophic microorganisms. Amplicons were pyrosequenced, and a total of 88,706 quality filtered reads were clustered into 1232 operational taxonomic units (OTU) across the six data sets. Comparisons of the taxonomic coverage achieved based on overlapping assignment of OTUs revealed that half of the eukaryotic taxa identified were missed by the universal eukaryotic barcoding marker. There were only little differences in OTU richness observed between organic- (farmyard manure), mineral- and nonfertilized soils. However, the community compositions appeared to be strongly structured by organic fertilization in all data sets other than that generated using the universal eukaryotic 18S rRNA gene primers, whereas mineral fertilization had only a minor effect. In addition, a co-occurrence based network analysis revealed complex potential interaction patterns between OTUs from different trophic levels, for example between fungivorous flagellates and fungi. Our results demonstrate that changes in pH, moisture and organic nutrients availability caused shifts in the composition of eukaryotic microbial communities at multiple trophic levels.

Communities of microbial eukaryotes in the mammalian gut within the context of environmental eukaryotic diversity.

Eukaryotic microbes (protists) residing in the vertebrate gut influence host health and disease, but their diversity and distribution in healthy hosts is poorly understood. Protists found in the gut are typically considered parasites, but many are commensal and some are beneficial. Further, the hygiene hypothesis predicts that association with our co-evolved microbial symbionts may be important to overall health. It is therefore imperative that we understand the normal diversity of our eukaryotic gut microbiota to test for such effects and avoid eliminating commensal organisms. We assembled a dataset of healthy individuals from two populations, one with traditional, agrarian lifestyles and a second with modern, westernized lifestyles, and characterized the human eukaryotic microbiota via high-throughput sequencing. To place the human gut microbiota within a broader context our dataset also includes gut samples from diverse mammals and samples from other aquatic and terrestrial environments. We curated the SILVA ribosomal database to reflect current knowledge of eukaryotic taxonomy and employ it as a phylogenetic framework to compare eukaryotic diversity across environment. We show that adults from the non-western population harbor a diverse community of protists, and diversity in the human gut is comparable to that in other mammals. However, the eukaryotic microbiota of the western population appears depauperate. The distribution of symbionts found in mammals reflects both host phylogeny and diet. Eukaryotic microbiota in the gut are less diverse and more patchily distributed than bacteria. More broadly, we show that eukaryotic communities in the gut are less diverse than in aquatic and terrestrial habitats, and few taxa are shared across habitat types, and diversity patterns of eukaryotes are correlated with those observed for bacteria. These results outline the distribution and diversity of microbial eukaryotic communities in the mammalian gut and across environments.

Optimal Eukaryotic 18S and Universal 16S/18S Ribosomal RNA Primers and Their Application in a Study of Symbiosis

Eukaryotic 18S ribosomal RNA (rRNA) gene primers that feature a wide coverage are critical in detecting the composition of eukaryotic microscopic organisms in ecosystems. Here, we predicted 18S rRNA primers based on consecutive conserved sites and evaluated their coverage efficiency and scope of application to different eukaryotic groups. After evaluation, eight of them were considered as qualified 18S primers based on coverage rate. Next, we examined common conserved regions in prokaryotic 16S and eukaryotic 18S rRNA sequences to design 16S/18S universal primers. Three 16S/18S candidate primers, U515, U1390 and U1492, were then considered to be suitable for simultaneous amplification of the rRNA sequences in three domains. Eukaryotic 18S and prokaryotic 16S rRNA genes in a sponge were amplified simultaneously using universal primers U515 and U1390, and the subsequent sorting of pyrosequenced reads revealed some distinctive communities in different parts of the sample. The real difference in biodiversity between prokaryotic and eukaryotic symbionts could be discerned as the dissimilarity between OTUs was increased from 0.005 to 0.1. A network of the communities in external and internal parts of the sponge illustrated the co-variation of some unique microbes in certain parts of the sponge, suggesting that the universal primers are useful in simultaneous detection of prokaryotic and eukaryotic microbial communities.

Eukaryotic microbial communities in hypersaline soils and sediments from the alkaline hypersaline Huama Lake as revealed by 454 pyrosequencing

In hypersaline ecosystems, microbial assemblages are structurally distinctive and play important roles in many microbiological and ecological processes. Here, eukaryotic microorganisms in hypersaline samples were investigated by 454 pyrosequencing of internal transcribed spacer (ITS) gene libraries. In total, 4,645, 1,677, and 5,912 reads were obtained from ITS libraries of waterlogged samples, salt crusts, and saline loess from the alkaline Huama Lake in Shaanxi, China. Analyses of pyrosequencing data revealed that the dominant genera were Dunaliella, Alternaria and Chlamydomonas, which dominated the microbial assemblages in the waterlogged sediments, the salt crusts and the saline loess from the lake banks, respectively. The various infrequent species were not commonly shared by the three types of samples, demonstrating that the eukaryotic microbial compositions of the different environments were distinct. However, the micro-eukaryotic assemblages associated with similar environmental conditions shared some components and were phylogenetically related. The eukaryotic microbial community composition was correlated with the pH value of the site (p=0.001; r(2)=0.99), but not with the concentration of total nitrogen or the inorganic ions investigated in this study. The results of this study demonstrated that the hypersaline ecosystems hosted surprisingly diverse eukaryotic microbial community.