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      Characterization of the Microbial Population Inhabiting a Solar Saltern Pond of the Odiel Marshlands (SW Spain)

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      Marine Drugs
      MDPI
      halo-extremophyles, archaea, 16S rRNA metagenomics, haloenzymes, Odiel marshlands

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          Abstract

          The solar salterns located in the Odiel marshlands, in southwest Spain, are an excellent example of a hypersaline environment inhabited by microbial populations specialized in thriving under conditions of high salinity, which remains poorly explored. Traditional culture-dependent taxonomic studies have usually under-estimated the biodiversity in saline environments due to the difficulties that many of these species have to grow at laboratory conditions. Here we compare two molecular methods to profile the microbial population present in the Odiel saltern hypersaline water ponds (33% salinity). On the one hand, the construction and characterization of two clone PCR amplified-16S rRNA libraries, and on the other, a high throughput 16S rRNA sequencing approach based on the Illumina MiSeq platform. The results reveal that both methods are comparable for the estimation of major genera, although massive sequencing provides more information about the less abundant ones. The obtained data indicate that Salinibacter ruber is the most abundant genus, followed by the archaea genera, Halorubrum and Haloquadratum. However, more than 100 additional species can be detected by Next Generation Sequencing (NGS). In addition, a preliminary study to test the biotechnological applications of this microbial population, based on its ability to produce and excrete haloenzymes, is shown.

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          Most cited references60

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          De novo metagenomic assembly reveals abundant novel major lineage of Archaea in hypersaline microbial communities.

          This study describes reconstruction of two highly unusual archaeal genomes by de novo metagenomic assembly of multiple, deeply sequenced libraries from surface waters of Lake Tyrrell (LT), a hypersaline lake in NW Victoria, Australia. Lineage-specific probes were designed using the assembled genomes to visualize these novel archaea, which were highly abundant in the 0.1-0.8 μm size fraction of lake water samples. Gene content and inferred metabolic capabilities were highly dissimilar to all previously identified hypersaline microbial species. Distinctive characteristics included unique amino acid composition, absence of Gvp gas vesicle proteins, atypical archaeal metabolic pathways and unusually small cell size (approximately 0.6 μm diameter). Multi-locus phylogenetic analyses demonstrated that these organisms belong to a new major euryarchaeal lineage, distantly related to halophilic archaea of class Halobacteria. Consistent with these findings, we propose creation of a new archaeal class, provisionally named 'Nanohaloarchaea'. In addition to their high abundance in LT surface waters, we report the prevalence of Nanohaloarchaea in other hypersaline environments worldwide. The simultaneous discovery and genome sequencing of a novel yet ubiquitous lineage of uncultivated microorganisms demonstrates that even historically well-characterized environments can reveal unexpected diversity when analyzed by metagenomics, and advances our understanding of the ecology of hypersaline environments and the evolutionary history of the archaea.
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            Industrial and environmental applications of halophilic microorganisms.

            In comparison with the thermophilic and the alkaliphilic extremophiles, halophilic microorganisms have as yet found relatively few biotechnological applications. Halophiles are involved in centuries-old processes such as the manufacturing of solar salt from seawater and the production of traditional fermented foods. Two biotechnological processes involving halophiles are highly successful: the production of beta-carotene by the green alga Dunaliella and the production of ectoine (1,4,5,6-tetrahydro-2-methyl-4-pyrimidinecarboxylic acid), used as a stabilizer for enzymes and now also applied in cosmetic products, from moderately halophilic bacteria. The potential use of bacteriorhodopsin, the retinal protein proton pump of Halobacterium, in optoelectronic devices and photochemical processes is being explored, and may well lead to commercial applications in the near future. Demand for salt-tolerant enzymes in current manufacturing or related processes is limited. Other possible uses of halophilic microorganisms such as treatment of saline and hypersaline wastewaters, and the production of exopolysaccharides, poly-beta-hydroxyalkanoate bioplastics and biofuel are being investigated, but no large-scale applications have yet been reported.
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              Environmental diversity of bacteria and archaea.

              The microbial way of life spans at least 3.8 billion years of evolution. Microbial organisms are pervasive, ubiquitous, and essential components of all ecosystems. The geochemical composition of Earth's biosphere has been molded largely by microbial activities. Yet, despite the predominance of microbes during the course of life's history, general principles and theory of microbial evolution and ecology are not well developed. Until recently, investigators had no idea how accurately cultivated microorganisms represented overall microbial diversity. The development of molecular phylogenetics has recently enabled characterization of naturally occurring microbial biota without cultivation. Free from the biases of culture-based studies, molecular phylogenetic surveys have revealed a vast array of new microbial groups. Many of these new microbes are widespread and abundant among contemporary microbiota and fall within novel divisions that branch deep within the tree of life. The breadth and extent of extant microbial diversity has become much clearer. A remaining challenge for microbial biologists is to better characterize the biological properties of these newly described microbial taxa. This more comprehensive picture will provide much better perspective on the natural history, ecology, and evolution of extant microbial life.
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                Author and article information

                Journal
                Mar Drugs
                Mar Drugs
                marinedrugs
                Marine Drugs
                MDPI
                1660-3397
                12 September 2018
                September 2018
                : 16
                : 9
                : 332
                Affiliations
                Laboratory of Biochemistry and Molecular Biology, Faculty of Experimental Sciences, Marine International Campus of Excellence (CEIMAR), University of Huelva, 21071 Huelva, Spain; patgomvil@ 123456gmail.com (P.G.-V.); vigara@ 123456uhu.es (J.V.)
                Author notes
                [* ]Correspondence: rleon@ 123456uhu.es ; Tel.: +34-959-219-951
                Article
                marinedrugs-16-00332
                10.3390/md16090332
                6164061
                30213145
                8e094c4c-0c0d-4038-81f6-1420202ef57a
                © 2018 by the authors.

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( http://creativecommons.org/licenses/by/4.0/).

                History
                : 28 June 2018
                : 08 September 2018
                Categories
                Article

                Pharmacology & Pharmaceutical medicine
                halo-extremophyles,archaea,16s rrna metagenomics,haloenzymes,odiel marshlands

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