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International audience; Les communautés bactériennes associées aux racines dePanicum turgide, exposés à des conditions arides, ont été investis-associée à une combinaison d'approches culturelles et métataxonomiques. Technologie traditionnelle basée sur la culture-ont été utilisées et 32 ​​isolats de racines irradiées ont été identifiés comme appartenant àActinobactéries, Bacteroidetes, Firmicutes et Proteobacteria embranchements. Quatre souches actinobactériennes ont étéRésistant aux rayonnements ionisants (IR) :Microbactériesp. PT8 (4,8 kGy (kGy)),Microcoquesp.PT11 (4,4 kGy),Kocuria rhizophilaPT10 (2,9 kGy) etPromicromonospora paniciPT9T(2,6 kGy), basésur le Ddixdose nécessaire pour une réduction de 90 % des unités formant colonies (UFC). Concernant l'enquêtedes communautés microbiennesin situ, analyses métataxonomiques de la diversité des microorganismes résistants aux IRassociée aux racines irradiées a révélé une dominance marquée des Actinobactéries (46,6%) etProtéobactéries (31,5%) par rapport aux Bacteroidetes (4,6%) et Firmicutes (3,2%). Irradiation gamma nonn'ont fait que modifier la structure des communautés bactériennes, mais ont également affecté leurs propriétés fonctionnelles.Des analyses comparatives des profils métaboliques ont indiqué l'induction de plusieurs voies liées à l'adaptation.au stress oxydatif dans les racines irradiées, comme la réparation de l'ADN, la synthèse de métabolites secondaires, la réactionenzymes atténuant les espèces oxygénées (ROS),etc. P. turgidumest emblématique des plantes adaptées au désert.Jusqu'à présent, aucun autre travail n'a porté sur le profil microbien des racines irradiées de cexérophyte.

Purpose: To assess exopolysaccharides (EPS) of Bacillus siamensis CV5, isolated from irradiated roots of Cistanche violacea, for their induction by ionizing radiation (IR) and their antioxidant and...

Simple Summary In the current investigation, we determine the archaeal diversity from halite-crystal salts sampled from Chott Djerid and Chott Douz, two saline Chotts in the Tunisian Sahara, using both culture-dependent and independent approaches citing DGGE (V3 regions), 16S rRNA-gene (full length gene ~ 1500 bp) based clone libraries and high-throughput sequencing technology of Illumina MiSeq platform (V3–V4 regions). The amalgamation of all results yielded a comprehensive view of the archaeal diversity represented by members of Halobacteria and Nanohaloarchaea classes. Abstract A thorough assessment of the phylogenetic diversity and community structure of halophilic archaea from three halite-crystal salts, processed from two separated saline systems of Southern Tunisia has been performed using culture dependent and independent methods targeting different regions of 16S rRNA gene sequences including DGGE, 16S rRNA clone libraries and Illumina Miseq sequencing. Two samples, CDR (red halite-crystal salts) and CDW (white halite-crystal salts), were collected from Chott-Eljerid and one sample CDZ (white halite-crystal salts) from Chott Douz. Fourteen isolates were identified as Halorubrum, Haloferax, Haloarcula, and Halogeometricum genera members. Culture-independent approach revealed a high diversity of archaeal members present in all samples, represented by the Euryarchaeal phylum and the dominance of the Halobacteria class. Nanohaloarchaea were also identified only in white halite samples based on metagenomic analysis. In fact, a total of 61 genera were identified with members of the Halorhabdus, Halonotius, Halorubrum, Haloarcula, and unclassified. Halobacteriaceae were shared among all samples. Unexpected diversity profiles between samples was observed where the red halite crust sample was considered as the most diverse one. The highest diversity was observed with Miseq approach, nevertheless, some genera were detected only with 16S rRNA clone libraries and cultured approaches.

14 páginas, 5 tablas, 4 figuras, A new strain belonging to the genus Kocuria, designed PT10, was isolated from irradiated roots of the xerophyte Panicum turgidum. Isolate PT10 is a Gram-positive, coccoid, aerobic and ionizing-radiation (IR)-resistant acti-nobacterium. PT10 has shown an ability to survive under extreme conditions, such as gamma irradiation, desiccation and high concentration of hydrogen peroxide. Phenotypic, chemotaxonomic and comparative genome analyses support the assignment of strain PT10 (LMG 31102 =DSM 108617) as Kocuria rhizophila. The complete genome sequence of PT10 consists of one chromosome (2,656,287 bps), with a 70.7% G +C content and comprises 2481 protein-coding sequences. A total of 1487 proteins were identified by LC-MS/MS profiling. In silico analyses revealed that the proteome of the oxidation-tolerant PT10 possesses several features explaining its IR-resistant phenotype and many adaptive pathways implicated in response to environmental pressures — desiccation, cold, reactive oxygen species and other stressors., This work was performed under the auspices of Université Lyon 1, UMR 5557, Ecologie Microbienne (France) and the National Center for Nuclear Sciences and Technologies CNSTN (MESRS, Tunisia) and in the ambit of the BIODESERT research program of the LR11-ES31 (BVBGR, ISBST, University of Manouba (MESRS, Tunisia). The LABGeM (CEA/ Genoscope & CNRS UMR8030), the France G ́enomique and French Bioinformatics Institute national infrastructures (funded as part of Investissement d’Avenir program managed by Agence Nationale pour la Recherche, contracts ANR-10-INBS-09 and ANR-11-INBS-0013) are acknowledged for support within the MicroScope annotation platform. We thank Betty Bigai, Corinne Sannaire and platform Genomique Environnementale PGE, UMR CNRS 5557 for technical support

4 tablas y 4 figuras., A novel strain of the genus Promicromonospora, designated PT9T, was recovered from irradiated roots of the xerophyte Panicum turgidum collected from the Ksar Ghilane oasis in southern Tunisia. Strain PT9T is aerobic, non-spore-forming, Gram- positive actinomycete that produces branched hyphae and forms white to yellowish-white colonies. Chemotaxonomic features, including fatty acids, whole cell sugars and polar lipid profles, support the assignment of PT9T to the genus Promicromonospora. The genomic relatedness indexes based on DNA-DNA hybridization and average nucleotide identity values revealed a signifcant genomic divergence between strain PT9T and all sequenced type strains of the taxon. Phylogenomic analysis showed that isolate PT9T was most closely related to Promicromonospora soli CGMCC 4.7398T. Phenotypic and phylogenomic analyses suggest that isolate PT9T represents a novel species of the genus Promicromonospora, for which the name Promicromonospora panici sp. nov. is proposed. The type strain is PT9T (LMG 31103T= DSM 108613T).The isolate PT9T is an ionizing-radiation-resistant actinobacterium (D10 value=2.6 kGy), with resistance to desiccation and hydrogen peroxide. The complete genome sequence of PT9T consists of 6,582,650 bps with 71.2% G+C content and 6291 protein-coding sequences. This genome will help to decipher the microbial genetic bases for ionizing-radiation resistance mechanisms including the response to oxidative stress., This work was performed under the auspices of Université Lyon 1, CNRS UMR 5557, Écologie Microbienne (France) and the National Center for Nuclear Sciences and Technology CNSTN (Tunisia) and in the ambit of the BIODESERT research program of the LR11-ES31 (BVBGR, ISBST, University of Manouba). The LABGeM (CEA/Genoscope & CNRS UMR8030), the France Génomique and French Bioinformatics Institute national infrastructures (funded as part of Investissement d'Avenir program managed by Agence Nationale pour la Recherche, contracts ANR-10-INBS-09 and ANR-11-INBS-0013) are acknowledged for support within them MicroScope annotation platform.

Bacterial communities associated with roots of

Phosphogypsum (PG) is a by-product of the phosphate fertilizer industry that represents an environmental concern. BRM17, a newly isolated strain from Tunisian PG BRM17 with the ability to solubilize inorganic phosphate, was identified as Pantoea sp. based on the analysis of 16S rDNA sequencing. The plant growth-promoting traits of BRM17 were analyzed by investigating the production of siderophores, indolic compounds (IAA), exopolysaccharides (EPS), ammonia (NH3), and 1-aminocyclopropane-1-carboxylate (ACC) deaminase activity. Observation of Arabidopsis thaliana roots inoculated with BRM17 tagged with a far-red fluorescent protein (mkate2) revealed that Pantoea sp. colonized the surface and inner part of the roots. A pot experiment was conducted to investigate the ability of BRM17 to promote the growth of canola (Brassica napus) in PG-amended soil in both sterilized and non-sterilized conditions. An increase in shoot and root biomass was observed in plants that were inoculated with BRM17 and cultivated in 2% PG amended soil. This effect was more significant in the sterilized condition. Furthermore, canola inoculated with BRM17 was able to grow in 100% PG, unlike non-inoculated plants. This study provides a strong basis for the further development of Pantoea sp. BRM17 inoculum to solubilize inorganic phosphate, due to its ability to overcome difficulties related to plant growth in PG dump sites or in PG-amended soils.

In this work, a native exopolysaccharide (nEPS) produced by Halomonas desertis G11 isolated from a Tunisian extreme environment was modified by gamma irradiation. Characterization as well as the antioxidant and antitumor activities of nEPS and its gamma-irradiated derivatives (iEPSs) were comparatively evaluated. In vitro and in vivo antioxidant potentials were determined by using different methods and through different antioxidant enzymes. The antitumor activity was checked against a human colon cancer cell line. Analyses of the complete genome sequence were carried out to identify genes implicated in the production of nEPS. Thus, the genomic biosynthesis pathway and the export mechanism of nEPS were proposed. Analyses of irradiation data showed that iEPSs acquired new functional groups, lower molecular weights, and gained significantly (p &lt, 0.05) higher antioxidant and antitumor abilities compared with nEPS. These findings provide a basis for using iEPSs as novel pharmaceutical agents for human therapies.

This study explored the possible use of a microbial carboxymethylated sulfated heteroexopolysaccharide (CS-hEPS) as a potential anticancer agent. The investigation was carried out through antioxidant, antifatigue, and antiproliferative activities. Antioxidant potential including scavenging DPPH and hydroxyl radical activities and reducing power was evaluated. Antifatigue activity was determined by assessing the endurance of mice using the forced swimming test. Following 30 days of CS-hEPS oral treatment at different doses, biochemical parameters related to fatigue such as lactic dehydrogenase (LDH), serum urea nitrogen (SUN), and hepatic glycogen (HG) contents were measured. Antitumor activities were investigated against human cancer liver and myelogenous leukemia cells. Results showed that CS-hEPS possesses notable antioxidant, antifatigue, and antitumor effects. CS-hEPS significantly inhibited the proliferation of leukemia (86.6 ± 0.32%) and cancer liver (58.6 ± 0.43%) cells. CS-hEPS are promising natural antioxidant, antifatigue, and antitumor harmless adjuvant materials that could be applied in human cancer therapy.

International audience; Phosphogypsum (PG) is an acidic by-product from the phosphate fertilizer industry and it is characterized by a low nutrient availability and the presence of radionuclides and heavy metals which pose a serious problem in its management. Here, we have applied Illumina MiSeq sequencing technology and five bioinformatics pipelines to explore the phylogenetic communities in Tunisian PG. Taking One Codex as a reference method, we present the results of 16S-rDNA-gene-based metataxonomics abundances with four other alternative bioinformatics pipelines (MetaGenome Rapid Annotation using Subsystem Technology (MG-RAST), mothur, MICrobial Community Analysis (MICCA) and Quantitative Insights into Microbial Ecology (QIIME)), when analyzing the Tunisian PG. Importantly, based on 16S rDNA datasets, the functional capabilities of microbial communities of PG were deciphered. They suggested the presence of PG autochthonous bacteria valorizable into (1) removal of radioactive elements and toxic heavy metals, (2) promotion of plant growth, (3) oxidation and (4) reduction of sulfate. These bacteria can be explored further for applications in the bioremediation of by-products, like PG, by different processes.

The presence of non-steroidal anti-inflammatory drugs, such as diclofenac (DCF), in the environment, is an emerging problem due to their harmful effects on non-target organisms, even at low concentrations. We studied the biodegradation of DCF by the strain D15 of Enterobacter hormaechei. The strain was isolated from an activated sludge, and identified as E. hormaechei based on its physiological characteristics and its 16 S RNA sequence. Using HPTLC and GC-MS methods, we demonstrated that this strain metabolized DCF at an elimination rate of 52.8%. In the presence of an external carbon source (glucose), the elimination rate increased to approximately 82%. GC-MS analysis detected and identified one metabolite as 1-(2,6-dichlorophenyl)-1,3-dihydro-2H-indol-2-one; it was produced as a consequence of dehydration and lactam formation reactions.

International audience; The early Frankia-Alnus symbiotic molecular exchanges were analyzed in detail by protein and RNA omics. For this, Frankia cells were placed in the presence of Alnus roots but separated by a dialysis membrane for 64 h. The bacterial cells were then harvested and analyzed by high-throughput proteomics and transcriptomics (RNA-seq). The most upregulated gene clusters were found to be the potassium transporter operon kdp and an ABC transporter operon of uncharacterized function. The most upregulated proteins were found to be acyl dehydrogenases and the potassium transporter Kdp. These suggest a preadaptation to the impending stresses linked to the penetration into isotonic host tissues and a possible rearrangement of the membrane. Another cluster among the 60 most upregulated ones that comprised two cellulases and a cellulose synthase was conserved among the Frankia and other actinobacteria such as Streptomyces. Cellulase activity was detected on CMC all along the length of the root but not away from it. Frankia alni ACN14a was found to be unable to respire or grow on glucose as sole carbon source. The cellulose synthase was found active at the tip of hyphae in response to Alnus root exudates, resulting in a calcofluor stained tip.

Here, we report the genomic features and the bioremediation potential of Halomonas desertis G11, a new halophilic species which uses crude oil as a carbon and energy source and displays intrinsic resistance to salt stress conditions (optimum growth at 10% NaCl). G11 genome (3.96 Mb) had a mean GC content of 57.82%, 3622 coding sequences, 480 subsystems and 64 RNA genes. Annotation predicted 38 genes involved in osmotic stress including the biosynthesis of osmoprotectants glycine-betaine, ectoine and osmoregulated periplasmic glucans. Genome analysis revealed also the versatility of the strain for emulsifying crude oil and metabolizing hydrocarbons. The ability of G11 to degrade crude oil components and to secrete a glycolipid biosurfactant with satisfying properties was experimentally confirmed and validated. Our results help to explain the exceptional capacity of G11 to survive at extreme desertic conditions, and highlight the metabolic features of this organism that has biotechnological and ecological potentialities.

International audience; Actinorhizal plants are ecologically and economically important. Symbiosis with nitrogen-fixing bacteria allows these woody dicotyledonous plants to colonise soils under nitrogen deficiency, water-stress or other extreme conditions. However, proteins involved in xerotolerance of symbiotic microorganisms have yet to be identified. Here we characterise the polyethylene glycol (PEG)-responding desiccome from the most geographically widespread Gram-positive nitrogen-fixing plant symbiont, Frankia alni, by next-generation proteomics, taking advantage of a Q-Exactive HF tandem mass spectrometer equipped with an ultra-high-field Orbitrap analyser. A total of 2,052 proteins were detected and quantified. Under osmotic stress, PEG-grown F. alni cells increased the abundance of envelope-associated proteins like ABC transporters, mechano-sensitive ion channels and Clustered Regularly Interspaced Short Palindromic Repeats CRISPR-associated (cas) components. Conjointly, dispensable pathways, like nitrogen fixation, aerobic respiration and homologous recombination, were markedly down-regulated. Molecular modelling and docking simulations suggested that the PEG is acting on Frankia partly by filling the inner part of an up-regulated osmotic-stress large conductance mechanosensitive channel. Actinobacteria belonging to the genus Frankia do establish nitrogen-fixing nodular symbiosis with the roots of 23 angiosperm genera that are collectively called " actinorhizals " 1. These plants form root nodules in which Frankia fixes nitrogen, thus permitting them to thrive in pioneer soils poor in nitrogen and organic matter, such as glacial moraines, lava fields, forest burnouts or anthropogenic sites such as mine spoils or hydrodam dykes 2. Frankia establishes a symbiotic association with the roots of several dicotyledonous plants. The different Frankia lineages form a coherent cluster at the root of the aerobic actinobacteria phylum 3 , and F. alni in particular establishes symbiosis with alder (Alnus) and bayberry (Morella) species 4. The interaction has evolved over several million years with a sophisticated dialogue that does not imply acy-lated N-acetyl-glucosamine oligomeric Nod factors 5. The Frankia determinants of symbiosis are still poorly known, which is for the most part due to the lack of a genetic transformation system. Transcriptomics has shown genes coding for nitrogenase (nif), hydrogenase uptake (hup), hopanoids (shc, hpn), iron-sulfur (suf) clusters as among the most up-regulated 6. Proteomics has also been used to analyze the symbiosis 7 to identify up-regulated peptides and it has shown the presence of Nif, Hup, Suf, Hop proteins as expected but also several transporters, regulators and various proteins involved in stress responses.

The identification of specific amino acids (AAs) or groups of functionally important AA residues in ionizing radiation-resistant bacteria (IRRB) is an important challenge in understanding the biological basis of resistance to ionizing radiation (IR; X-rays and gamma-rays). To address this problem, we compared homologous sites in multiple alignments of proteins of IRRB and IR-sensitive bacteria (IRSB) using the DeltaProt Toolbox. Substitution patterns were used as evidence for selection of certain AAs over others. Our results show that, in contrast to aromatic AAs, small/tiny AAs tend to be preferred in IRRB compared to IRSB. In agreement with previous experimental data showing that oxidation of AA residues is causative in the killing of irradiated cells and that IR resistance is correlated with the accumulation of divalent manganese ions (Mn2+)–peptide–orthophosphate (Pi) complexes, we proposed a chemical interpretation based on the Hard and Soft (Lewis) Acids and Bases (HSAB) concept. These findings should assist future efforts in selecting mutations for rational design of proteins with enhanced IR tolerance properties.

Phosphogypsum (PG) is an industrial waste composed mainly by sulfate, turning it a suitable sulfate source for sulfate-reducing bacteria (SRB). In the present work, the capability of two SRB communities, one enriched from Portuguese PG (culture PG) and the other from sludge from a wastewater treatment plant (culture WWT-1), to use sulfate from PG was compared. In addition, the impact of this sulfate-rich waste in the microbial community was assessed. The highest efficiency in terms of sulfate reduction was observed with culture WWT-1. The bacterial composition of this culture was not significantly affected when sodium sulfate from the nutrient medium was replaced by PG as a sulfate source. Next generation sequencing (NGS) showed that this community was phylogenetically diverse, composed by bacteria affiliated to Clostridium, Arcobacter, and Sulfurospirillum genera and by SRB belonging to Desulfovibrio, Desulfomicrobium, and Desulfobulbus genera. In contrast, the bacterial structure of the community enriched from PG was modified when sodium sulfate was replaced by PG as the sulfate source. This culture, which showed the poorest performance in the use of sulfate from PG, was mainly composed by SRB related to Desulfosporosinus genus. The present work provides new information regarding the phylogenetic characterization of anaerobic bacterial communities with the ability to use PG as sulfate donor, thus, contributing to improve the knowledge of microorganisms suitable to be used in PG bioremediation. Additionally, this paper demonstrates that an alternative to lactate and low-cost carbon source (wine wastes) can be used efficiently for that purpose.

The physiological and global transcriptomic response of the metal-resistant bacterium Cupriavidus metallidurans CH34 to stable non-radioactive strontium (Sr) was investigated. C. metallidurans CH34 was able to survive and proliferate in the presence of relatively high concentrations of SrCl2 (concentration of Sr needed to inactivate 90 % of the cells = 70 mM; minimum inhibitory concentration = 120 mM). Gene expression in cells was analyzed after short-term (30 min) exposure to low (5 mM) and high (60 mM) concentrations of SrCl2. The transcription of the gene clusters annotated as hmyFCBA and czcCBADRS, coding for ion efflux pumps, was significantly induced following exposure to Sr relative to that in non-stressed CH34 cells. Sr precipitation was observed during aerobic growth of CH34 in the presence of 60 mM SrCl2. Transmission electron microscopy images of cells grown with Sr also showed very small solid-phase precipitates located in the vicinity of cells, but mainly extracellular. Environmental scanning electron microscopy with energy-dispersive X-ray spectrometry indicated that the sequestered extracellular solid-phase Sr was present in the form of strontium carbonate.

Stones in arid environments are inhabited by actinobacteria of the family Geodermatophilaceae like the genera Blastococcus and Modestobacter frequently isolated from altered calcarenites. Their habitat requires adaptation to light-induced and other stresses that generate reactive oxygen species. Here, we show that representative members of the species Blastococcus saxobsidens , Geodermatophilus obscurus , and Modestobacter multiseptatus are differentially adapted to stresses associated with arid environments. Whereas B. saxobsidens was found to be sensitive to gamma radiation (D10 = 900 Gy; 10% survival at 900 Gy), M. multiseptatus was moderately (D10 = 6000 Gy) and G. obscurus was highly tolerant (D10 = 9000 Gy). A difference in resistance to high-frequency (λ value = 254 nm) UV was shown by B. saxobsidens , M. multiseptatus , and G. obscurus , being sensitive, tolerant, and highly tolerant (D10 of 6, 900, and > 3500 kJ m−2, respectively). Tolerance to desiccation, mitomycin C and hydrogen peroxide correlated with the ionizing radiation and UV resistance profiles of the three species and were correlated with the pigments synthesized. Resistance to heavy metals/metalloids did not follow the same pattern, with resistance to Ag2+ and Pb2+ being similar for B. saxobsidens , M. multiseptatus , and G. obscurus , whereas resistance to ![Graphic][1] , Cr2+, or Cu2+ was greater for B. saxobsidens than for the other two species. The stress resistance profiles of M. multiseptatus and B. saxobsidens were reflected in different calcarenite colonization patterns. While M. multiseptatus was predominantly isolated from the first two millimeters of stone surface, B. saxobsidens was predominantly isolated from the deeper part of the stone where it is better protected from sun irradiation, suggesting that the response to light- and desiccation-induced oxidative stress is an important driver for niche colonization in the stone biotope. [1]: /embed/inline-graphic-1.gif

Aim: To assess esterase profiling of members of Geodermatophilaceae isolated from desert stones and monuments in Tunisia and Egypt. Methods and Results: Members of Geodermatophilaceae family isolated from desert stones and monuments in Tunisia and Egypt were characterized by partial 16S rRNA sequences. Twenty-five strains were clustered in three dissimilar groups of the genera Geodermatophilus (12 strains), Blastococcus (5 strains) and Modestobacter (3 strains). Isolates were also screened and typed based on major groups of esterase hydrolytic activity. Their esterase patterns were determined and compared to those of ten reference strains belonging to Geodermatophilaceae family. Strains exhibited a diverse and complex pattern of electrophoretic esterase bands, and 31 haplotypes were obtained for the 35 investigated strains. Esterases produced by members of Geodermatophilaceae family have an optimal activity around 40°C and at pH 8. Esterases from Geodermatophilus strains display a high resistance to thermal inactivation and alkaline pH and retaining 30 and 20% of activity after heating for 20 min at 120°C and at pH 12, respectively, and were completely inactivated after 30 min at 120°C. Enzyme activity has been strongly activated in the presence of Ca2+and Mg2+ ions and moderately by Zn2+ and was markedly inhibited by Cu2+ and Co2+ ions. Conclusions: Geodermatophilaceae isolates share a rich and particular pool of esterase activities that could be directly linked to harsh conditions characterizing their ecological habitat including high level of aridity, temperature, ionic strength and low nutrient availability. Significance and Impact of the Study: Esterase could be considered as enzymatic signature that outlines adaptability of Geodermatophilaceae in arid area.

Deinococcus radiodurans possesses a DNA damage response mechanism that acts via the PprI protein to induce RecA and PprA proteins, both of which are necessary in conferring extreme radioresistance. In an effort to further delineate the nature of the DNA damage response mechanism in D. radiodurans, we set out to identify novel components of the PprI-dependent signal transduction pathway in response to radiation stress. Here we demonstrate the discovery of a novel regulatory protein, PprM (a modulator of the PprI-dependent DNA damage response), which is a homolog of cold shock protein (Csp). Disruption of the pprM gene rendered D. radiodurans significantly sensitive to gamma-rays. PprM regulates the induction of PprA but not that of RecA. PprM belongs in a distinct clade of a subfamily together with Csp homologs from D. geothermalis and Thermus thermophilus. Purified PprM is present as a homodimer under physiological conditions, as the case with Escherichia coli CspD. The pprA pprM double-disruptant strain exhibited higher sensitivity than the pprA or pprM single disruptant strains, suggesting that PprM regulates other hitherto unknown protein(s) important for radioresistance besides PprA. This study strongly suggests that PprM is involved in the radiation response mediated by PprI in D. radiodurans.

The Geodermatophilaceae are unique model systems to study the ability to thrive on or within stones and their proteogenomes (referring to the whole protein arsenal encoded by the genome) could provide important insight into their adaptation mechanisms. Here we report the detailed comparative genome analysis of Blastococcus saxobsidens (Bs), Modestobacter marinus (Mm) and Geodermatophilus obscurus (Go) isolated respectively from the interior and the surface of calcarenite stones and from desert sandy soils. The genome-scale analysis of Bs, Mm and Go illustrates how adaptation to these niches can be achieved through various strategies including ` molecular tinkering/ opportunism' as shown by the high proportion of lost, duplicated or horizontally transferred genes and ORFans. Using high-throughput discovery proteomics, the three proteomes under unstressed conditions were analyzed, highlighting the most abundant biomarkers and the main protein factors. Proteomic data corroborated previously demonstrated stone-related ecological distribution. For instance, these data showed starvation-inducible, biofilm-related and DNA-protection proteins as signatures of the microbes associated with the interior, surface and outside of stones, respectively.

The extremely radioresistant eubacterium Deinococcus radiodurans and the phenotyp‐ ically related prokaryotes, whose genomes have been completely sequenced, are presently used as model species in several laboratories to study the lethal effects of DNA-damaging and protein-oxidizing agents, particularly the effects of ionizing radiation (IR). Unfortunately, providing relevant information about radioresistant prokaryotes (RP) in a neatly centralized and organized manner still remains a need. In this study, we designed RadioP1 Web resource (www.radiop.org.tn) to gather information about RP defined by the published literature with specific emphasis on (i) predicted genes that produce and protect against oxidative stress, (ii) predicted proteins involved in DNA repair mechanisms and (iii) potential uses of RP in biotechnology. RadioP1 allows the complete RP proteogenomes to be queried using various patterns in a user-friendly and interactive manner. The output data can be saved in plain text, Excel or HyperText Markup Language (HTML) formats for subsequent analyses. Moreover, RadioP1 provides for users a tool “START ANALY‐ SIS”, including the previously described R-packages “drc” and “lethal”, to generate exponential or sigmoid survival curves with D10 and D50 values. Furthermore, when accessible, links to external databases are provided. Supplementary data will be included in the future when the sequences of other RP genomes will become available.

A novel γ-radiation-resistant and Gram-staining-positive actinobacterium designated BMG 8133T was isolated from a limestone collected in the Sahara desert of Tunisia. The strain produced dry, pale-pink colonies with an optimum growth at 35–40 °C and pH 6.5–8.0. Chemotaxonomic and molecular characteristics of the isolate matched those described for members of the genus Geodermatophilus. The peptidoglycan contained meso-diaminopimelic acid as diagnostic diamino acid. The main polar lipids were phosphatidylcholine, diphosphatidylglycerol, phosphatidylinositol, phosphatidylethanolamine and one unspecified glycolipid. MK-9(H4) was the dominant menaquinone. Galactose and glucose were detected as diagnostic sugars. The major cellular fatty acids were branched-chain saturated acids iso-C16 : 0 and iso-C15 : 0. The DNA G+C content of the novel strain was 74.5 %. The 16S rRNA gene sequence showed highest sequence identity with Geodermatophilus ruber (98.3 %). Based on phenotypic results and 16S rRNA gene sequence analysis, strain BMG 8133T is proposed to represent a novel species, Geodermatophilus sabuli sp. nov. The type strain is BMG 8133T ( = DSM 46844T = CECT 8820T).

The taxonomic position of an aerobic actinobacterial strain, BMG841(T), isolated from the Bulla Regia monument (Tunisia) and exhibiting a high resistance to gamma-radiation (D10 ~9 kGy) was determined using polyphasic approach. The optimal growth range was found to be 25-35 °C at pH of 7.0-8.5. The strain was observed to form black dry colonies. Chemotaxonomic characteristics of the isolate showed a cell wall type III, with galactose and glucose as diagnostic sugars; phosphatidylcholine, phosphatidylinositol, diphosphatidylglycerol, phosphatidylethanolamine and an unidentified glycolipid as main polar lipids; and MK-9(H4) as the predominant menaquinone. The major cellular fatty acids were identified as iso-C16:0 and iso-C15:0. Phylogenetic analysis indicated that strain BMG841(T) represents a novel member of the genus Geodermatophilus with high 16S rRNA gene sequence identity with Geodermatophilus saharensis (98.28 %). Based on phylogenetic and phenotypic analysis, strain BMG841(T) is proposed as the type strain (=DSM 46841(T) = CECT 8821(T)) of a novel species, Geodermatophilus bullaregiensis.

An orange-black, Gram-positive, aerobic and gamma-ray resistant actinobacterium was isolated from the ruins of a Roman aqueduct located in Northern Tunisia. The optimal growth for the strain was found to be at 25-35 °C and at pH 6.0-9.5. Chemotaxonomic and molecular characteristics of the isolate matched those described for members of the genus Geodermatophilus. The peptidoglycan was found to contain meso-diaminopimelic acid as diagnostic diaminoacid. The main polar lipids were identified as phosphatidylcholine, diphosphatidylglycerol, phosphatidylinositol, phosphatidylethanolamine, an unidentified glycolipid and an unidentified aminophospholipid; MK-9(H4) was found to be the dominant menaquinone and galactose was detected as the diagnostic sugar, with glucose, ribose and mannose also present. The major cellular fatty acids were identified as branched-chain saturated acids iso-C16:0, iso-C15:0 and iso-H-C16:0. The 16S rRNA gene showed 95.4-99.6 % sequence identity with the type strains of the genus Geodermatophilus. DNA-DNA relatedness values with closely related species were 39.9 ± 4.9, 33.9 ± 1.9, 27.0 ± 2.5 and 13.2 ± 1.35 % with Geodermatophilus amargosae, G. normandii, G. saharensis and G. tzadiensis respectively. Based on phenotypic results and 16S rRNA gene sequence analysis, strain BMG801(T) (=DSM 46834(T) = CECT 8822(T)) is proposed to represent the type strain of a novel species, Geodermatophilus aquaeductus sp. nov.

Catalase plays a key role in protecting cells against toxic reactive oxygen species. Here we report on the cloning, purification and characterization of a catalase (KatA, DR1998) from the extremely radioresistant bacterium Deinococcus radiodurans. The size of purified D. radiodurans KatA monomer was 65 kDa while gel filtration revealed that the size of the enzyme was 240 kDa, suggesting that KatA formed a homotetramer in solution. Purified KatA displayed a final specific activity of 68,800 U/mg of protein. The catalase activity of KatA was inhibited by sodium azide, sodium cyanide and 3-amino-1,2,4-triazole. The absorption spectrum of KatA exhibited a Soret band at 408 nm. The position of the spectral peak remained unchanged following reduction of KatA with dithionite. No peroxidase activity was found for KatA. These results demonstrate that D. radiodurans KatA is a typical monofunctional heme-containing catalase. The stability of KatA with respect to H2O2 stress was superior to that of commercially available Aspergillus niger and bovine liver catalases. The relative abundance of KatA in cells in addition to the H2O2 resistance property may play a role in the survival strategy of D. radiodurans against oxidative damage.

RecA is a highly conserved bacterial protein that plays crucial roles in many cellular processes and hence is a potential target in the chemotherapy of bacterial infections. An understanding of the functional similarity between RecA proteins from different bacterial species should yield further insights into the biochemistry of RecA protein, along with the potential for new approaches to facilitate the improvement of RecA-targeted drugs. In this technical note, the authors present an in silico method based on tri-oligonucleotide usage correlations (TOUC) to predict the functional similarity between two RecA orthologs. The TOUC values analyzed in this study are in good agreement with the available experimental results. This method should prove useful in guiding future experimental efforts aimed at furthering our understanding of the biochemistry of RecA proteins and subsequent development of new drugs that modulate RecA biological activities in bacteria.

A gamma radiation-resistant, Gram reaction-positive, aerobic and chemoorganotrophic actinobacterium, initially designated Geodermatophilus obscurus subsp. dictyosporus G-5(T), was not validly named at the time of initial publication (1968). G-5(T) formed black-colored colonies on GYM agar. The optimal growth range was 25-35 °C, at pH 6.5-9.5 and in the absence of NaCl. Chemotaxonomic and molecular characteristics of the isolate matched those described for members of the genus Geodermatophilus. The DNA G + C content of the strain was 75.3 mol%. The peptidoglycan contained meso-diaminopimelic acid as diagnostic diamino acid. The main polar lipids were phosphatidylcholine, diphosphatidylglycerol, phosphatidylinositol, phosphatidylethanolamine and one unspecified glycolipid; MK-9(H4) was the dominant menaquinone and galactose was detected as a diagnostic sugar. The major cellular fatty acids were branched-chain saturated acids, iso-C16:0 and iso-C15:0. The 16S rRNA gene showed 94.8-98.4 % sequence identity with the members of the genus Geodermatophilus. Based on phenotypic results and 16S rRNA gene sequence analysis, strain G-5(T) is proposed to represent a novel species, Geodermatophilus dictyosporus and the type strain is G-5(T) (=DSM 43161(T) = CCUG 62970(T) = MTCC 11558(T) = ATCC 25080(T) = CBS 234.69(T) = IFO 13317(T) = KCC A-0154(T) = NBRC 13317(T)). The INSDC accession number is HF970584.

The γ-irradiation effects on polyphenolic content and antioxidant capacity of parathion-pretreated leaves of Salvia officinalis plant were investigated. The analysis of phenolic extracts of sage without parathion showed that irradiation decreased polyphenolic content significantly (p

At the outset, I believe that the concept of ionizing-radiation (IR) resistance needs to be clarified in a tangible manner for readers of this chapter. I propose the following general definition adopted in a previous paper (Sghaier et al., 2008): An ionizing-radiation-resistant prokaryotes (IRRP) is any vegetative prokaryote that can thrive after exposure to high, acute IR (generally, with a D10 value the dose necessary to effect a 90% reduction in Colony Forming Units greater than 1 kGy) using efficient physiological, genetic and proteic protection and repair mechanisms to fully amend its DNA DSBs. IR resistance has been observed in a broad range of prokaryotic groups (Kopylov et al., 1993), including hyperthermophilic Archaea (P. abyssi, P. furiosus, Thermococcus marinus, Thermococcus radiotolerans and Thermococcus gammatolerans) (DiRuggiero et al., 1997; Jolivet et al., 2003a; Jolivet et al., 2003b; Jolivet et al., 2004), halophilic Archaea (Halobacterium sp.) (Kottemann et al., 2005), the Deinococcus-Thermus group (many Deinococcus sp. and Truepera radiovictrix) (Albuquerque et al., 2005), Actinobacteria (Rubrobacter radiotolerans, Rubrobacter xylanophilus and Kineococcus radiotolerans) (Yoshinaka et al., 1973; Ferreira et al., 1999; Phillips et al., 2002; Chen et al., 2004), Proteobacteria (Methylobacterium radiotolerans and Acinetobacter radioresistens) (Ito and Iizuka, 1971; Nishimura et al., 1994), Cyanobacteria (Chroococcidiopsis sp.) (Billi et al., 2000), and Sphingobacteria (Hymenobacter actinosclerus) (Collins et al., 2000). However, with the exception of Deinococcus and Pyrococcus, very little information is available regarding the mechanisms of IR resistance and comparative genomics in the above-mentioned prokaryotes. D. radiodurans is the current gold-medallist of IR resistance among prokaryotes with a completely sequenced genome (Liolios et al., 2006; Liolios et al., 2010), and can amend more than 100 DSBs per chromosome, induced by IR, without loss of viability (Moseley, 1983; White et al., 1999). After breaking of its 3.2 Mb genome into 20−30 kb pieces by a high dose of IR, D. radiodurans fascinatingly reassembles its genome such that 3 hr later fully restructured nonrearranged chromosomes are present (Galhardo and Rosenberg, 2009). Nine interrelated explanations for the extreme IR resistance of D. radiodurans have been proposed: (i) the binding of IrrI (DR0171) to genomic repeat sequences that might prevent exhaustive chromosomal degradation after IR exposure irr for IR resistance (Udupa et al., 1994), (ii) the

Interactomes of proteins under positive selection from ionizing-radiation-resistant bacteria (IRRB) might be a part of the answer to the question as to how IRRB, particularly Deinococcus radiodurans R1 (Deira), resist ionizing radiation. Here, using the Database of Interacting Proteins (DIP) and the Protein Structural Interactome (PSI)-base server for PSI map, we have predicted novel interactions of orthologs of the 58 proteins under positive selection in Deira and other IRRB, but which are absent in IRSB. Among these, 18 domains and their interactomes have been identified in DNA checkpoint and repair; kinases pathways; energy and nucleotide metabolisms were the important biological processes that were found to be involved. This finding provides new clues to the cellular pathways that can to be important for ionizing-radiation resistance in Deira.

The extremely radioresistant bacterium Deinococcus radiodurans contains two LexA homologues (LexA1 and LexA2) that are possible transcriptional regulators associated with the DNA damage response. In this study, resequencing revealed that there was an additional cytosine nucleotide (nucleotide position 612) in the D. radiodurans lexA2 gene. Purified LexA2 possessed proteolytic activity that could be stimulated by RecA. In an effort to gain an insight into the role of LexA2 in the radiation response mechanism, recA, lexA1 and lexA2 disruptant strains were generated and investigated. The intracellular level of RecA increased in lexA1 and lexA2 disruptant strains following γ-irradiation as in the wild-type strain. These results indicated that the two LexA homologues did not possess functional overlap regarding the induction of RecA. The lexA2 disruptant strains exhibited a much higher resistance to γ-rays than the wild-type strain. Furthermore, a luciferase assay showed that pprA promoter activation was enhanced in the lexA2 disruptant strain following γ-irradiation. The pprA gene encoding the novel radiation-inducible protein PprA plays a critical role in the radioresistance of D. radiodurans. The increase in radioresistance of the lexA2 disruptant strain is explained in part by the enhancement of pprA promoter activation.