Micro-eukaryotic diversity in hypolithons from Miers Valley, Antarctica

The discovery of extensive and complex hypolithic communities in both cold and hot deserts has raised many questions regarding their ecology, biodiversity and relevance in terms of regional productivity. However, most hypolithic research has focused on the bacterial elements of the community. This study represents the first investigation of micro-eukaryotic communities in all three hypolith types. Here we show that Antarctic hypoliths support extensive populations of novel uncharacterized bryophyta, fungi and protists and suggest that well known producer-decomposer-predator interactions may create the necessary conditions for hypolithic productivity in Antarctic deserts

A novel bacterial Water Hypersensitivity-like protein shows in vivo protection against cold and freeze damage

Metagenomic library screening, by functional or sequence analysis, has become an established method for the identification of novel genes and gene products, including genetic elements implicated in microbial stress response and adaptation. We have identified, using a sequence based approach, a fosmid clone from an Antarctic desert soil metagenome library containing a novel gene which codes for a protein homologous to a Water Hypersensitive domain (WHy). The WHy domain is typically found as a component of specific LEA (Late Embryogenesis Abundant) proteins, particularly the LEA-14 (LEA-8) variants, which occur widely in plants, nematodes, bacteria and archaea and which are typically induced by exposure to stress conditions. The novel Why-like protein, (165 amino acid, 18.6 kDa) exhibits a largely invariant NPN motif at the N-terminus and has high sequence identity to genes identified in Pseudomonas genomes. Expression of this protein in E. coli significantly protected the recombinant host against cold and freeze stress.South African National Antarctic Program of the National Research Foundation.http://femsle.oxfordjournals.org2016-08-31hb201

Normalization of environmental metagenomic DNA enhances the discovery of under-represented microbial community members

Normalization is a procedure classically employed to detect rare sequences in cellular expression profiles (i.e. cDNA libraries). Here, we present a normalization protocol involving the direct treatment of extracted environmental metagenomic DNA with S1 nuclease; referred to as Normalization of metagenomic DNA: NmDNA. We demonstrate that NmDNA, prior to post hoc PCR based experiments (16S rRNA gene T-RFLP fingerprinting and clone library), increased the diversity of sequences retrieved from environmental microbial communities by detection of rarer sequences. This approach could be used to enhance the resolution of detection of ecologically relevant rare members in environmental microbial assemblages.National Research Foundation (NRF) of South Africa (BlueSkies program 81693).http://onlinelibrary.wiley.com/journal/10.1111/(ISSN)1472-765X2016-04-30hb201

Draft genome sequence of the Antarctic polyextremophile Nesterenkonia sp. strain AN1

Nesterenkonia sp. strain AN1 was isolated from Antarctic soil and is a polyextremophile, being tolerant of low temperatures, high salt concentrations, and high alkalinity. Here we report the draft genome sequence of this strain.This project was partially supported by the National Research Foundation– South African National Antarctic Research Programme (NRFSANAP) (award 80256) and the Genomics Research Institute (GRI), University of Pretoria.http://genomea.asm.org/am201

Evidence of variability in the structure and recruitment of rhizospheric and endophytic bacterial communities associated with arable sweet sorghum (Sorghum bicolor (L) Moench)

Sorghum is the second most cultivated crop in Africa and is a staple food source of many African communities. Exploiting the associated plant growth promoting bacteria (PGPB) has potential as an agricultural biotechnology strategy to enhance sorghum growth, yield and nutritional properties. Here we use Terminal-Restriction Fragment Length Polymorphism (TRFLP) and Denaturing Gradient Gel Electrophoresis (DGGE) to evaluate the factors that potentially shape rhizospheric and endophytic bacterial communities associated with sorghum farmed in South Africa. Microbial diversity was typically higher in the rhizosphere and rhizoplane compared to the endophytic zones (root, shoot and stem). Geographical location was one of the main drivers in describing microbial community assemblages found in rhizospheric and endophytic sorghum-linked niches. NO3-N, total nitrogen and pH were clearly identified as the main abiotic factors shaping sorghum-associated soil communities. Our results also suggest that specific bacterial taxa with potential N-fixing capacities (Acetobacter sp., Azospirillum sp., Pantoea sp., Bacillus sp. and cyanobacteria) are consistently detected in sorghum-created rhizospheric and endophytic environments, irrespective of environmental factor effects.The South African National Research Foundation (NRF)http://link.springer.com/journal/11104hb2016Genetic

Engineering pyruvate decarboxylase-mediated ethanol production in the thermophilic host Geobacillus thermoglucosidasius

This study reports the expression, purification and kinetic characterization of a PDC from Gluconobacter oxydans. Kinetic analyses showed the enzyme to have high affinity for pyruvate (120μM at pH 5), high catalytic efficiency (4.75 x 105 M-1s-1 at pH 5), a pHopt of approximately 4.5 and an in vitro temperature optimum at approximately 55°C (the highest yet reported for a bacterial PDC). Due to good in vitro thermostablity (approximately 40% enzyme activity retained after 30 minutes at 65°C) this PDC was considered to be a suitable candidate for heterologous expression in the thermophile Geobacillus thermoglucosidasius. Initial studies using a variety of methods failed to detect activity at any growth temperature. However, the application of codon harmonization (i.e., mimicry of the heterogeneous host’s transcription and translational rhythm) yielded a protein that was fully functional in the thermophilic strain at 45°C (as determined by enzyme activity, Western blot, mRNA detection and ethanol productivity). Here we describe the successful expression of PDC in a true thermophile. Yields as high as 0.35 g/g ±0.04 ethanol per gram of glucose consumed were detected, highly competitive to those reported in ethanologenic thermophilic mutants. Although activities could not be detected at temperatures approaching the growth optimum for the strain, this study highlights that the possibility that previously unsuccessful expression of pdcs in Geobacillus spp. may be the result of ineffective transcription / translation coupling.National Research Foundation South Africahttp://link.springer.com/journal/253hb201

Evidence of species recruitment and development of hot desert hypolithic communities

Hypoliths, photosynthetic microbial assemblages found underneath translucent rocks, are widely distributed within the western region of the Namib Desert and other similar environments. Terminal restriction fragment length polymorphism (T-RFLP) analysis was used to assess the bacterial community structure of hypoliths and surrounding soil (below and adjacent to the hypolithic rock) at a fine scale (10 m radius). Multivariate analysis of T-RFs showed that hypolithic and soil communities were structurally distinct. T-RFLP-derived operational taxonomic units were linked to 16S rRNA gene clone libraries. Applying the ecological concept of ‘indicator species’, six and nine indicator lineages were identified for hypoliths and soil, respectively. Hypolithic communities were dominated by cyanobacteria affiliated to Pleurocapsales, whereas actinobacteria were prevalent in the soil. These results are consistent with the concept of species sorting and suggest that the bottom of the quartz rocks provides conditions suitable for the development of discrete and demonstrably different microbial assemblages. However, we found strong evidence for neutral assembly processes, as almost 90% of the taxa present in the hypoliths were also detected in the soil. These results suggest that hypolithons do not develop independently from microbial communities found in the surrounding soil, but selectively recruit from local populations.Innovation Fund UID 71682(PhD. scholarship for TPM), the National Research Fund of South Africa and the University of the Western Cape (T.P.M, A.V, I.M.T, and D.A.C). D.C.L. and S.B.P. were funded by the Hong Kong Research Grants Council (Grant number HKU7733/08HKU7763/10).http://onlinelibrary.wiley.com/journal/10.1111/(ISSN)1758-2229hb201

Evidence of novel plant-species specific ammonia oxidizing bacteria clades in acidic South African fynbos soils

Ammonia-oxidizing bacteria (AOB) are essential in the biogeochemical cycling of nitrogen as they catalyze the rate-limiting oxidation of ammonia into nitrite. Since their first isolation in the late 19th century, chemolithoautotrophic AOBs have been identified in a wide range of natural (e.g., soils, sediments, estuarine, and freshwaters) and man created or impacted habitats (e.g., wastewater treatment plants and agricultural soils). However, little is known on the plant-species association of AOBs, particularly in the nutrient-starved fynbos terrestrial biome. In this study, we evaluated the diversity of AOBs in the plant canopy of three South African fynbos-specific plant species, namely Leucadendron xanthoconus, Leucospermum truncatulum and Leucadendron microcephalum, through the construction of amoA-gene clone libraries. Our results clearly demonstrate that plant-species specific and monophyletic AOB clades are present in fynbos canopy soils.Claude Leon Foundation, the South African National Research Foundation (NRF) and University of Pretoria.http://onlinelibrary.wiley.com/journal/10.1002/(ISSN)1521-40282016-08-28hb201

Niche-dependent genetic diversity in Antarctic metaviromes

The metaviromes from 2 different Antarctic terrestrial soil niches have been analyzed. Both hypoliths (microbial assemblages beneath transluscent rocks) and surrounding open soils showed a high level diversity of tailed phages, viruses of algae and amoeba, and virophage sequences. Comparisons of other global metaviromes with the Antarctic libraries showed a niche-dependent clustering pattern, unrelated to the geographical origin of a given metavirome. Within the Antarctic open soil metavirome, a putative circularly permuted, »42kb dsDNA virus genome was annotated, showing features of a temperate phage possessing a variety of conserved protein domains with no significant taxonomic affiliations in current databases.National Research Foundation (South Africa) and the Genomics Research Institute of the University of Pretoria (South Africa).http://www.tandfonline.com/loi/kbac202015-12-31hb201

Metagenomic analysis of the viral community in Namib desert hypoliths

Hypolithic microbial communities are specialized desert communities inhabiting the underside of translucent rocks where they are sheltered from harsh environmental conditions. Here, we present the first study of the viral fraction of these communities isolated from the hyperarid Namib Desert (coastal South Western Africa). Using next-generation sequencing of the isolated viral fraction, the diversity and taxonomic composition of hypolith communities was mapped and a functional assessment of the sequences determined. Phylotypic analysis showed that bacteriophages belonging to the order Caudovirales with the family Siphoviridae were most prevalent. A major fraction of phage types was linked by database homologies to Bacillus or Geobacillus sp. as a host. Phylogenetic analyses of terL and phoH marker genes indicated that many of the sequences were novel and distinct from known isolates and environments, an observation supported by the class distribution of identified ribonucleotide reductases. The composition of the viral hypolith fraction was not completely consistent with Namib hypolith phylotypic surveys, in which the cyanobacterial genus Chroococcidiopsis was found to be dominant. This could be attributed to lacking sequence information about hypolith viruses/bacteria in public databases or the hypothesis that hypolithic communities actively recruit viruses from the surrounding open soil in which Bacillaceae-infecting phages are more commonly found.http://onlinelibrary.wiley.com/journal/10.1111/(ISSN)1462-2920hb201