Phenylobacterium glaciei sp. nov., isolated from Vestrebroggerbreen, a valley glacier in Svalbard, Arctic.

A Gram-stain-negative, rod-shaped bacterial strain designated as 20VBR1T was isolated from a valley glacier (Vestrebroggerbreen) snout ice sample from Ny-Ålesund, Svalbard, Arctic. The colonies were smooth, circular and light creamish on half-strength R2A agar and grew at 10-35 °C (optimum, 20 °C), at pH 6.5-8.0 (optimum, 7.0) and with 0-2.5 % (w/v) NaCl (optimum, 0.5 %). 16S rRNA gene sequence analysis revealed that strain 20VBR1T belonged to the genus Phenylobacterium and was most closely affiliated to Phenylobacterium aquaticum W2-3-4T (97.65 % similarity), Phenylobacterium haematophilum LMG 11050T (97.57 %) and Phenylobacterium koreense Slu-01T (96.91 %). 20VBR1T has a genome size of 4.24 Mb, comprising 4185 predicted genes with a DNA G+C content of 67.86 mol%. DNA-DNA hybridization experiments indicated that the DNA-DNA relatedness between strain 20VBR1T and P. aquaticum KACC 18306T was 41.95±4.36 %, well below the threshold (<70 %) to delineate bacterial species. Genome relatedness indexes revealed that the average nucleotide identity and digital DNA-DNA hybridization values between 20VBR1T and its closest phylogenomic relative, P. aquaticum KACC 18306T, were 78.97 and 22.10 %, respectively. The predominant isoprenoid quinone was ubiquinone (Q-10) and the major polar lipids were phosphatidylglycerol, one unknown phospholipid, one unknown glycolipid and four unidentified polar lipids. The major fatty acids (>10 %) of strain 20VBR1T were summed feature 8 (comprising C18 : 1 ω7c and/or C18 : 1 ω6c), summed feature 3 (comprising C16 : 1 ω6c and/or C16 : 1 ω7c) and C16 : 0. Based on the physiological, biochemical, chemotaxonomic, phylogenetic and phylogenomic analyses, isolate 20VBR1T is considered to represent a novel species of the genus Phenylobacterium, for which the name Phenylobacterium glaciei sp. nov. is proposed. The type strain is 20VBR1T (=JCM 33227 T=DSM 111428 T=MCC 4220 T).

Complete genome sequence and comparative genome analysis of Alcanivorax sp. IO_7, a marine alkane-degrading bacterium isolated from hydrothermally-influenced deep seawater of southwest Indian ridge.

Genome of Alcanivorax sp. IO_7, an alkane degrading deep-sea bacteria isolated from hydrothermally-influenced Southwest Indian Ridge was sequenced and analysed. Genomic data mining revealed gene clusters for degrading n-alkane and cycloalkanes, including biosurfactant production. The strain was shown to grow on hexadecane as its sole carbon source, supporting the findings of genomic analysis. Presence of cyclohexanone monooxygenase among genomic islands suggest that this strain may have used gene transfer to enhance its hydrocarbon degradation ability. Genes encoding for heavy metal resistance, multidrug resistance and multiple natural product biosynthesis crucial for survival in the hydrothermally influenced deep sea environment were detected. In our comparative genome analysis, it was evident that marine Alcanivorax strains contain a suite of enzymes for n-alkane and haloalkanoate degradation. Comparative genome and genomic synteny analysis provided insights into the physiological features and adaptation strategies of Alcanivorax sp. IO_7 in the deep-sea hydrothermal environment.

Genomic insights into the molecular mechanisms of a Pseudomonas strain significant in its survival in Kongsfjorden, an Arctic fjord.

Whole-genome sequence of Pseudomonas sp. Kongs-67 retrieved from Kongsfjorden, an Arctic fjord, has been investigated to understand the molecular machinery required for microbial association and survival in a polar fjord. The genome size of Kongs-67 was 4.5 Mb and was found to be closely related to the Antarctic P. pelagia strain CL-AP6. This genome encodes for chemotaxis response regulator proteins (CheABB1RR2VWYZ), chemoreceptors (methyl-accepting chemotaxis proteins), and flagellar system proteins (FliCDEFGOPMN, FlhABF, FlgBCDEFGHIJKL, and MotAB proteins) vital in cellular interactions in the dynamic fjord environment. A high proportion of genes were assigned to biofilm formation (pgaABCD operon) and signal transduction protein categories (EnvZ/OmpR, CpxA/CpxR, PhoR/PhoB, PhoQ) indicating that the biofilm formation in Kongs-67 could be tightly regulated in response to the availability of signalling-metabolites. The genome of Kongs-67 encoded for HemBCD, CbiA, CobABNSTOQCDP, and BtuBFR proteins involved in cobalamin biosynthesis and transport along with proteins for siderophore-mediated iron channelling (PchR, Fur protein, FpvA); crucial in a microbial association. The genomes of Arctic strain Kongs-67 and Antarctic strain CL-AP6 were similar which is indicative of retainment of the core genes in the polar Pseudomonas strains that could be vital in conferring evolutionary adaptation for its survival in a polar fjord. Thus, our study contributes to the knowledge on the genetics of a polar Pseudomonas member exhibiting biosynthetic potentials and suggest Pseudomonas sp. Kongs-67 as a suitable candidate for the investigation of functional aspects of molecular adaptations in the polar marine environment.

Roseitranquillus sediminis gen. nov., sp. nov. a novel genus and species of the family Rhodobacteraceae, isolated from sediment of an Arctic fjord.

A Gram-negative, aerobic, non-motile, oxidase-positive, catalase-positive, rod-shaped bacterium, designated strain MCCB 386T was isolated from sediment samples collected from Kongsfjorden, an Arctic fjord. The strain MCCB 386T showed growth at 4-37 °C (optimum 27°C) in the presence of 1-8% NaCl (w/v, optimum 3.5%) and at pH 6.0-8.0 (optimum pH 7.0). The major fatty acids were C18:1ω7c (54.0%) and 11-methyl C18:1ω7c (22.6%). The dominant respiratory quinone was Q-10. The major polar lipids comprised of phosphatidylcholine (PC), diphosphatidylglycerol (DPG), phosphatidylglycerol (PG), phosphoglycolipid (PGL), one unidentified aminolipid, two glycolipids and two unidentified lipids. The genomic G+C content of the strain MCCB 386T was 68.1 mol%. The 16 S rRNA gene sequences based phylogenetic analysis of MCCB 386T showed that Psychromarinibacter halotolerans YBW34T (95.88%) is the most closely related species. In addition, overall genome relatedness indices (OGRI) of MCCB 386T with closely related strains were lower than threshold level for species and genus delineation. The analysis of Biosynthetic Gene clusters (BGCs) revealed the potential of this strain for production of novel bioactive secondary metabolites. As per polyphasic taxonomic characterisation, strain MCCB 386T represents a novel species of a novel genus for which the name Roseitranquillus sediminis gen. nov., sp. nov. is suggested. The type strain of the species is MCCB 386T (= JCM 33,538T= KACC 21,531T).

Pseudonocardia cytotoxica sp. nov., a novel actinomycete isolated from an Arctic fjord with potential to produce cytotoxic compound.

Herein we report the isolation of a novel actinomycete, strain MCCB 268T, from the sediment sample collected from a high Arctic fjord Kongsfjorden. MCCB 268T showed greater than 97% 16S rRNA gene sequence similarity with those of Pseudonocardia konjuensis LM 157T (98.06%), Pseudonocardia soli NW8-21 (97.22%) Pseudonocardia endophytica YIM 56035 (97.08%) and Pseudonocardia nantongensis KLBMP 1282 (97.34%) showing that the strain should be assigned to the genus Pseudonocardia. DNA-DNA hybridization with Pseudonocardia konjuensis LM 157T showed only 41.5% relatedness to strain MCCB 268T. The whole genome of the strain MCCB 268T was sequenced. Whole-genome average nucleotide identity, dDDH (%) and genome tree analysis demonstrated that strain significantly differed from other Pseudonocardia species. The G + C content was 70.5 mol%. MCCB 268T exhibited in vitro cytotoxicity and through bioassay guided fractionation followed by HPLC separation a cytotoxic compound (I) was isolated. The compound (I) was identified as 1-acetyl-ß-carboline through NMR spectra and high-resolution mass spectrometry. Compound (I) showed cytotoxicity against lung cancer cell line and mode of anticancer activity was found to be through the induction of apoptosis. Based on the genotypic and phenotypic features, MCCB 268T ought to be classified as a novel species under the genus Pseudonocardia for which the name Pseudonocardia cytotoxica sp. nov. is proposed (= CCUG72333T = JCM32718T).

Alteromonas pelagimontana sp. nov., a marine exopolysaccharide-producing bacterium isolated from the Southwest Indian Ridge.

A novel exopolysaccharide-producing strain, designated as 5.12T, was isolated from a sediment sample from the Southwest Indian Ridge, Indian Ocean. The strain was Gram-stain-negative, motile, strictly aerobic, and oxidase- and catalase-positive. It grew optimally at 35 °C, at pH 6.0 and in the presence of 3.5 % (w/v) NaCl. Its major isoprenoid quinone was ubiquinone-8 (Q-8) and summed feature 3 (comprising C16 : 1ω6c and/or C16 : 1ω7c), C16 : 0 and C18 : 1ω7c were the major cellular fatty acids. The DNA G+C content was 46.1 mol%. 16S rRNA gene sequence analysis suggested that strain 5.12T is a member of the genus Alteromonas. Strain 5.12T exhibited close 16S rRNA gene sequence similarity to Alteromonas lipolytica JW12T (96.1 %), Alteromonas hispanica F-32T (95.9 %), Alteromonas confluentis DSSK2-12T (95.9 %), Alteromonas litorea TF-22T (95.6 %) and Alteromonas mediterranea DET (95.5 %). Strain 5.12T contained phosphatidylethanolamine and phosphatidylglycerol as the major polar lipids. Owing to significant differences in the 16S rRNA gene sequences, as well as the phenotypic and chemotaxonomic characteristics, the novel isolate described here merits classification as a representative of a novel species of the genus Alteromonas, for which the name Alteromonas pelagimontana sp. nov. is proposed. The type strain of this species is 5.12T (LMG 29661T= MCC 3250T).

Spatio-Temporal Monitoring and Ecological Significance of Retrievable Pelagic Heterotrophic Bacteria in Kongsfjorden, an Arctic Fjord.

Kongsfjorden is a glacial fjord in the Arctic that is influenced by both Atlantic and Arctic water masses. In the present report retrievable heterotrophic bacteria isolated from two distinct zones (outer and inner fjord) of Kongsfjorden was studied during summer to fall of 2012. 16S rRNA gene sequences of the retrievable heterotrophic bacteria corresponded to γ-proteobacteria (13 phylotypes), α-proteobacteria (3 phylotypes), Bacteroidetes (4 phylotypes) and Actinobacteria (2 phylotypes). The heterotrophic bacterial community structure was fundamentally different in different months which could be linked to changes in the water masses and/or phytoplankton bloom dynamics. It is hypothesized that monitoring the retrievable heterotrophic bacterial assemblage in the fjord would give valuable insights into the complex ecological role they play under extreme and dynamic conditions.

Microbial diversity and associated metabolic potential in the supraglacial habitat of a fast-retreating glacier: a case study of Patsio glacier, North-western Himalaya.

In the present study, we investigated the microbial community composition and their associated metabolic potentials using the 16S rRNA gene (V3-V4) and ITS (ITS1) amplicon sequencing approach in the Patsio glacier. The bacterial community composition was mainly dominated by Bacteroidota (18%-38% of total reads) and Cyanobacteria (9%-30%), along with a rare Candidate phylum Patescibacteria. Ferruginibacter (13%) and Polaromonas (8%) were the most dominant genera identified across the samples known to have potential ecological roles in colonization, driving the functioning of supraglacial habitats. The prevalence of metabolic genes associated with nitrogen, carbon and sulfur cycling processes was identified in the present study. The fungal diversity was dominated by members of unclassified phyla, followed by Ascomycota (up to 6%) and Basidiomycota (up to 4%), in terms of its relative abundance. The relative abundance of Fusarium and Didymella (8%-14%) was higher among the high altitude, cryoconite samples (P1-P5), while Rhodotorula (12%-29%) dominated in the glacial ice debris samples (P6-P8). Thus, our study provides significant insights into dynamics of microbial communities and its potential ecological roles in the changing climate.

Complete genome sequence of Alteromonas pelagimontana 5.12T, a marine exopolysaccharide-producing bacterium isolated from hydrothermally influenced deep-sea sediment of eastern Southwest Indian Ridge.

The whole genome of Alteromonas pelagimontana 5.12T, a psychrotolerant deep-sea bacterium isolated from the sediment sample of eastern Southwest Indian Ridge, was sequenced and analysed for understanding its metabolic capacities and biosynthesis potential of natural products. The circular genome contained 4.3 Mb with a GC content of 42.6 mol%. Genomic data mining revealed a gene cluster for heavy metal resistance (czcABC, acrB, arsR1, copA, nikA, mntH, mntP), exopolysaccharides (EPS; epsCDEFHLM) and polyhydroxyalkanoates (PHA; phbC) production, as well as genes involved in complex polysaccharide degradation. Genes that could allow strain 5.12T to cope with acid stress (ibaG) and heat shock (ibpA, hslR) were observed along with ten chaperone-encoding genes which could possibly play vital role in adaptability of this strain to the hydrothermally influenced environment. Gene clusters for secondary metabolite production such as bacteriocin and arylpolyene were also predicted. Thus, genome sequencing and data mining provided insights into the molecular mechanisms involved in the adaptation to hydrothermally influenced deep-sea environment that could promote further experimental exploration.

Bacterial community structure of a glacio-marine system in the Arctic (Ny-Ålesund, Svalbard).

The bacterial community composition of a valley glacier in Svalbard, its pro-glacial channels, and the associated downstream fjord ecosystem was investigated so as to figure out the degree to which downslope transport of microbes from the glacier systems along a hydrological continuum impose an effect on the patterns of diversity in the fjord system. A combination of culture based and high-throughput amplicon sequencing approach was followed which resulted in significant variation (R = 0.873, p = 0.001) in the bacterial community structure between these ecosystems. Dominance of sequences belonging to class ß-Proteobacteria was seen in the glacier snow, ice and melt waters (MW) while a relatively higher abundance of OTUs belonging to α-Proteobacteria and Verrucomicrobiae demarcated the fjord waters. Similarity percentage (SIMPER) analysis of the Operational Taxonomic Units (OTUs) showed that OTU 1,105,280 (9.15%) and OTU 331 (6.5%) belonging to Burkholderiaceae (ß-proteobacteria) and OTU 101,660 (5.76%) and OTU 520 (5.07%) belonging to Rhodobacteraceae (α-proteobacteria) contributed maximum to the overall dissimilarity between the sampling sites. The bacterial community from the MWs were found to be true signatures of the glacier ecosystem while the Kongsfjorden bacterial fraction mostly represented heterotrophic marine taxa influenced by warm Atlantic waters and presence of organic matter. Significant presence of unknown taxa in the MWs suggests the need to study such unexplored, transient niches for a better understanding of the associated microbial processes. Among the various environmental parameters measured, nutrients (NO3- and SiO42-) were found to exhibit strong association with the MW bacterial community while temperature, trace metals, Cl- and SO42- ions were found to influence the fjord bacterial community. The significant differences in the bacterial community composition between the glacier and the fjord ecosystem suggest the unique nature of these systems which in turn is influenced by the associated environmental parameters.

Draft Genome Sequence of Idiomarina sp. Strain 5.13, a Highly Stress-Resistant Bacterium Isolated from the Southwest Indian Ridge.

Idiomarina sp. strain 5.13, able to produce biopolymer and exopolysaccharide, was isolated from a sediment sample collected from the Southwest Indian Ridge, Indian Ocean. Analysis of its draft genome sequence provides insights into its remarkable stress tolerance and offers the genetic basis for harnessing the biotechnological potential of this strain.

Diversity of retrievable heterotrophic bacteria in Kongsfjorden, an Arctic fjord.

The diversity and abundance of retrievable pelagic heterotrophic bacteria in Kongsfjorden, an Arctic fjord, was studied during the summer of 2011 (June, August, and September). Retrievable bacterial load ranged from 103 to 107CFUL-1 in June, while it was 104-106CFUL-1 in August and September. Based on 16S rRNA gene sequence similarities, a higher number of phylotypes was observed during August (22 phylotypes) compared to that during June (6 phylotypes) and September (12 phylotypes). The groups were classified into four phyla: Firmicutes, Actinobacteria, Proteobacteria, and Bacteroidetes. Bacteroidetes was represented only by a single member Leewenhoekiella aequorea during the three months and was dominant (40%) in June. However, this dominance changed in August to a well-known phytopathogenic species Rhodococcus fascians (32%), which could be a result of decrease in the phytoplankton biomass following the secondary bloom. It is the first report of Halomonas titanicae isolation from the Arctic waters. It showed an increase in its abundance with the intrusion of Atlantic water into Kongsfjorden. Increased abundance of Psychrobacter species in the late summer months coincided with the presence of cooler waters. Thus, the composition and function of heterotrophic bacterial community was fundamentally different in different months. This could be linked to the changes in the water masses and/or phytoplankton bloom dynamics occurring in Arctic summer.

Streptomyces artemisiae MCCB 248 isolated from Arctic fjord sediments has unique PKS and NRPS biosynthetic genes and produces potential new anticancer natural products.

After screening marine actinomycetes isolated from sediment samples collected from the Arctic fjord Kongsfjorden for potential anticancer activity, an isolate identified as Streptomyces artemisiae MCCB 248 exhibited promising results against the NCI-H460 human lung cancer cell line. H460 cells treated with the ethyl acetate extract of strain MCCB 248 and stained with Hoechst 33342 showed clear signs of apoptosis, including shrinkage of the cell nucleus, DNA fragmentation and chromatin condensation. Further to this treated cells showed indications of early apoptotic cell death, including a significant proportion of Annexin V positive staining and evidence of DNA damage as observed in the TUNEL assay. Amplified PKS 1 and NRPS genes involved in secondary metabolite production showed only 82% similarity to known biosynthetic genes of Streptomyces, indicating the likely production of a novel secondary metabolite in this extract. Additionally, chemical dereplication efforts using LC-MS/MS molecular networking suggested the presence of a series of undescribed tetraene polyols. Taken together, these results revealed that this Arctic S. artemisiae strain MCCB 248 is a promising candidate for natural products drug discovery and genome mining for potential anticancer agents.

Diversity of retrievable heterotrophic bacteria in Kongsfjorden, an Arctic fjord

Abstract The diversity and abundance of retrievable pelagic heterotrophic bacteria in Kongsfjorden, an Arctic fjord, was studied during the summer of 2011 (June, August, and September). Retrievable bacterial load ranged from 103 to 107 CFU L−1 in June, while it was 104-106 CFU L−1 in August and September. Based on 16S rRNA gene sequence similarities, a higher number of phylotypes was observed during August (22 phylotypes) compared to that during June (6 phylotypes) and September (12 phylotypes). The groups were classified into four phyla: Firmicutes, Actinobacteria, Proteobacteria, and Bacteroidetes. Bacteroidetes was represented only by a single member Leewenhoekiella aequorea during the three months and was dominant (40%) in June. However, this dominance changed in August to a well-known phytopathogenic species Rhodococcus fascians (32%), which could be a result of decrease in the phytoplankton biomass following the secondary bloom. It is the first report of Halomonas titanicae isolation from the Arctic waters. It showed an increase in its abundance with the intrusion of Atlantic water into Kongsfjorden. Increased abundance of Psychrobacter species in the late summer months coincided with the presence of cooler waters. Thus, the composition and function of heterotrophic bacterial community was fundamentally different in different months. This could be linked to the changes in the water masses and/or phytoplankton bloom dynamics occurring in Arctic summer.