scholarly journals Culture Enrichment Combined With Long-Read Sequencing Facilitates Genomic Understanding of Hadal Sediment Microbes

2021 ◽  
Vol 8 ◽  
Author(s):  
Huan Wang ◽  
Meng Wang ◽  
Shen Fan ◽  
Jie Lu ◽  
Yi Lan ◽  
...  
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Carbon Sources ◽  
Nitrite Reduction ◽  
Rrna Gene ◽  
Carbohydrate Utilization ◽  
Microbial Species ◽  
Culture Enrichment ◽  
Long Read ◽  
Microbial Proliferation

Culture enrichment was developed to discover the uncharted microbial species in the environmental microbiota. Yet this strategy has not been widely used to study microbes of deep-sea environments. Here, we report the cultivation and metagenomic analysis of oceanic sediment microbiota collected from 6,477 m deep in the Mariana Trench. The sediment samples were cultured anaerobically in the laboratory for 4 months, before being subjected to full-length 16S rRNA gene sequencing using the PacBio technique and metagenome sequencing using both the Illumina and Oxford Nanopore techniques. The 16S rRNA gene analyses revealed 437 operational taxonomic units specific to the cultured microbes, despite the lower diversity of the cultured microbiota in comparison with the original. Metagenome analyses revealed the prevalence of functions related to respiration, energy production, and stress response in the cultured microbes, suggesting these functions may contribute to microbial proliferation under laboratory conditions. Binning of the assembled metagenome contigs of the cultured microbiota generated four nearly complete genomes affiliated to yet unclassified species under the genera Alcanivorax, Idiomarina, Sulfitobacter, and Erythrobacter. Excepting Alcanivorax, the other three genera were almost undetectable in the original samples and largely enriched in the cultured samples. The four genomes possessed a variety of genes for carbohydrate utilization and nitrite reduction, pointing to an ability to respire diverse carbon sources using nitrite as the final electron acceptor. Taken together, the findings suggest that a combination of culture enrichment and long-read sequencing is an ideal way to mine novel microbial species in the hadal environment, particularly species that are rare in their native environmental niches, and thus expand our understanding of the hadal microbial diversity and function.

scholarly journals Phyllobacterium loti sp. nov. isolated from nodules of Lotus corniculatus

2014 ◽  
Vol 64 (Pt_3) ◽  
pp. 781-786 ◽  
Author(s):  
Maximo Sánchez ◽  
Martha-Helena Ramírez-Bahena ◽  
Alvaro Peix ◽  
María J. Lorite ◽  
Juan Sanjuán ◽  
...  
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Sequence Similarity ◽  
Carbon Sources ◽  
Lotus Corniculatus ◽  
Culture Conditions ◽  
Rrna Gene ◽  
Content Type ◽  
Link Type ◽  
The 16S Rrna Gene

Strain S658T was isolated from a Lotus corniculatus nodule in a soil sample obtained in Uruguay. Phylogenetic analysis of the 16S rRNA gene and atpD gene showed that this strain clustered within the genus Phyllobacterium . The closest related species was, in both cases, Phyllobacterium trifolii PETP02T with 99.8 % sequence similarity in the 16S rRNA gene and 96.1 % in the atpD gene. The 16S rRNA gene contains an insert at the beginning of the sequence that has no similarities with other inserts present in the same gene in described rhizobial species. Ubiquinone Q-10 was the only quinone detected. Strain S658T differed from its closest relatives through its growth in diverse culture conditions and in the assimilation of several carbon sources. It was not able to reproduce nodules in Lotus corniculatus. The results of DNA–DNA hybridization, phenotypic tests and fatty acid analyses confirmed that this strain should be classified as a representative of a novel species of the genus Phyllobacterium , for which the name Phyllobacterium loti sp. nov. is proposed. The type strain is S658T( = LMG 27289T = CECT 8230T).

scholarly journals Paucibacter toxinivorans gen. nov., sp. nov., a bacterium that degrades cyclic cyanobacterial hepatotoxins microcystins and nodularin

2005 ◽  
Vol 55 (4) ◽  
pp. 1563-1568 ◽  
Author(s):  
Jarkko Rapala ◽  
Katri A. Berg ◽  
Christina Lyra ◽  
R. Maarit Niemi ◽  
Werner Manz ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
16S Rrna ◽  
16S Rrna Gene ◽  
Gene Sequence ◽  
Sequence Similarity ◽  
Carbon Sources ◽  
16S Rrna Gene Sequence ◽  
Rrna Gene ◽  
Rrna Gene Sequence

Thirteen bacterial isolates from lake sediment, capable of degrading cyanobacterial hepatotoxins microcystins and nodularin, were characterized by phenotypic, genetic and genomic approaches. Cells of these isolates were Gram-negative, motile by means of a single polar flagellum, oxidase-positive, weakly catalase-positive and rod-shaped. According to phenotypic characteristics (carbon utilization, fatty acid and enzyme activity profiles), the G+C content of the genomic DNA (66·1–68·0 mol%) and 16S rRNA gene sequence analysis (98·9–100 % similarity) the strains formed a single microdiverse genospecies that was most closely related to Roseateles depolymerans (95·7–96·3 % 16S rRNA gene sequence similarity). The isolates assimilated only a few carbon sources. Of the 96 carbon sources tested, Tween 40 was the only one used by all strains. The strains were able to mineralize phosphorus from organic compounds, and they had strong leucine arylamidase and chymotrypsin activities. The cellular fatty acids identified from all strains were C16 : 0 (9·8–19 %) and C17 : 1 ω7c (<1–5·8 %). The other predominant fatty acids comprised three groups: summed feature 3 (<1–2·2 %), which included C14 : 0 3-OH and C16 : 1 iso I, summed feature 4 (54–62 %), which included C16 : 1 ω7c and C15 : 0 iso OH, and summed feature 7 (8·5–28 %), which included ω7c, ω9c and ω12t forms of C18 : 1. A more detailed analysis of two strains indicated that C16 : 1 ω7c was the main fatty acid. The phylogenetic and phenotypic features separating our strains from recognized bacteria support the creation of a novel genus and species, for which the name Paucibacter toxinivorans gen. nov., sp. nov. is proposed. The type strain is 2C20T (=DSM 16998T=HAMBI 2767T=VYH 193597T).

scholarly journals Clostridium lundense sp. nov., a novel anaerobic lipolytic bacterium isolated from bovine rumen

2006 ◽  
Vol 56 (3) ◽  
pp. 625-628 ◽  
Author(s):  
Dores G. Cirne ◽  
Osvaldo D. Delgado ◽  
Sankar Marichamy ◽  
Bo Mattiasson
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Rumen Fluid ◽  
Sequence Similarity ◽  
Carbon Sources ◽  
Rrna Gene ◽  
Strictly Anaerobic ◽  
Bovine Rumen ◽  
Hybridization Data ◽  
Clostridium Tetanomorphum

A strictly anaerobic, mesophilic, endospore-forming, lipolytic bacterium, designated strain R1T, was isolated from bovine rumen fluid and characterized. Cells of this isolate were Gram-positive, non-motile rods that formed spherical terminal spores. The overall biochemical and physiological characteristics indicated that this strain should be placed in the genus Clostridium. The strain grew at temperatures between 25 and 47 °C (optimum, 37 °C), at pH between 5·0 and 8·5 (optimum pH 5·5–7·0) and in NaCl concentrations of 0–3 % (w/v). The isolate was not able to utilize glucose or other carbohydrates as carbon sources. The DNA G+C content was 31·2 mol%. Sequence analysis of the 16S rRNA gene of R1T revealed that it has the closest match (98 % similarity) with Clostridium tetanomorphum DSM 4474T. The highest levels of DNA–DNA relatedness of the isolate were 61·9 and 54·3 % with Clostridium pascui DSM 10365T and C. tetanomorphum DSM 4474T, respectively. Based on 16S rRNA gene sequence similarity, phylogenetic analysis, DNA G+C content, DNA–DNA hybridization data and distinct phenotypic characteristics, strain R1T (=DSM 17049T=CCUG 50446T) was classified in the genus Clostridium, as a member of a novel species, for which the name Clostridium lundense sp. nov. is proposed.

scholarly journals Photobacterium rosenbergii sp. nov. and Enterovibrio coralii sp. nov., vibrios associated with coral bleaching

2005 ◽  
Vol 55 (2) ◽  
pp. 913-917 ◽  
Author(s):  
F. L. Thompson ◽  
C. C. Thompson ◽  
S. Naser ◽  
B. Hoste ◽  
K. Vandemeulebroecke ◽  
...  
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Type Strain ◽  
Gene Sequence ◽  
Sequence Similarity ◽  
Phylogenetic Analyses ◽  
Carbon Sources ◽  
16S Rrna Gene Sequence ◽  
Rrna Gene ◽  
Rrna Gene Sequence

Six new Vibrio-like isolates originating from different species of bleached and healthy corals around Magnetic Island (Australia) were investigated using a polyphasic approach. Phylogenetic analyses based on 16S rRNA, recA and rpoA gene sequences split the isolates in two new groups. Strains LMG 22223T, LMG 22224, LMG 22225, LMG 22226 and LMG 22227 were phylogenetic neighbours of Photobacterium leiognathi LMG 4228T (95·6 % 16S rRNA gene sequence similarity), whereas strain LMG 22228T was related to Enterovibrio norvegicus LMG 19839T (95·5 % 16S rRNA gene sequence similarity). The two new groups can be distinguished from closely related species on the basis of several phenotypic features, including fermentation of d-mannitol, melibiose and sucrose, and utilization of different compounds as carbon sources, arginine dihydrolase activity, nitrate reduction, resistance to the vibriostatic agent O/129 and the presence of fatty acids 15 : 0 iso and 17 : 0 iso. The names Photobacterium rosenbergii sp. nov. (type strain LMG 22223T=CBMAI 622T=CC1T) and Enterovibrio coralii sp. nov. (type strain LMG 22228T=CBMAI 623T=CC17T) are proposed to accommodate these new isolates. The G+C contents of the DNA of the two type strains are respectively 47·6 and 48·2 mol%.

scholarly journals Erratum to: Improved OTU-picking using long-read 16S rRNA gene amplicon sequencing and generic hierarchical clustering

Microbiome ◽  
2015 ◽  
Vol 3 (1) ◽  
Author(s):  
Oscar Franzén ◽  
Jianzhong Hu ◽  
Xiuliang Bao ◽  
Steven H. Itzkowitz ◽  
Inga Peter ◽  
...  
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Hierarchical Clustering ◽  
Amplicon Sequencing ◽  
Rrna Gene ◽  
Long Read

scholarly journals Methylibium petroleiphilum gen. nov., sp. nov., a novel methyl tert-butyl ether-degrading methylotroph of the Betaproteobacteria

2006 ◽  
Vol 56 (5) ◽  
pp. 983-989 ◽  
Author(s):  
Cindy H. Nakatsu ◽  
Krassimira Hristova ◽  
Satoshi Hanada ◽  
Xian-Ying Meng ◽  
Jessica R. Hanson ◽  
...  
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Gene Sequence ◽  
Carbon Sources ◽  
16S Rrna Gene Sequence ◽  
Methyl Tert Butyl Ether ◽  
Rrna Gene ◽  
Rrna Gene Sequence ◽  
Butyl Ether ◽  
The 16S Rrna Gene

A Gram-negative, rod-shaped, motile, non-pigmented, facultative aerobe that grew optimally at pH 6.5 and 30 °C (strain PM1T) was isolated for its ability to completely degrade the gasoline additive methyl tert-butyl ether. Analysis of the 16S rRNA gene sequence indicated that this bacterium was a member of the class Betaproteobacteria in the Sphaerotilus–Leptothrix group. The 16S rRNA gene sequence identity to other genera in this group, Leptothrix, Aquabacterium, Roseateles, Sphaerotilus, Ideonella and Rubrivivax, ranged from 93 to 96 %. The chemotaxonomic data including Q-8 as the major quinone, C16 : 1ω7c and C16 : 0 as the major fatty acids and a DNA G+C content of 69 mol%, support the inclusion of strain PM1T in the class Betaproteobacteria. It differed from other members of the Sphaerotilus–Leptothrix group by being a facultative methylotroph that used methanol as a sole carbon source, and by also being able to grow heterotrophically in defined media containing ethanol, toluene, benzene, ethylbenzene and dihydroxybenzoates as sole carbon sources. On the basis of the morphological, physiological, biochemical and genetic information, a new genus and species, Methylibium petroleiphilum gen. nov., sp. nov., is proposed, with PM1T (=ATCC BAA-1232T=LMG 22953T) as the type strain.

scholarly journals Improved OTU-picking using long-read 16S rRNA gene amplicon sequencing and generic hierarchical clustering

Microbiome ◽  
2015 ◽  
Vol 3 (1) ◽  
Author(s):  
Oscar Franzén ◽  
Jianzhong Hu ◽  
Xiuliang Bao ◽  
Steven H. Itzkowitz ◽  
Inga Peter ◽  
...  
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Hierarchical Clustering ◽  
Amplicon Sequencing ◽  
Rrna Gene ◽  
Long Read

scholarly journals Silvimonas terrae gen. nov., sp. nov., a novel chitin-degrading facultative anaerobe belonging to the ‘Betaproteobacteria’

2005 ◽  
Vol 55 (6) ◽  
pp. 2329-2332 ◽  
Author(s):  
Hee-Chan Yang ◽  
Wan-Taek Im ◽  
Dong-Shan An ◽  
Woo-shin Park ◽  
In S. Kim ◽  
...  
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Gene Sequence ◽  
Sequence Similarity ◽  
Carbon Sources ◽  
16S Rrna Gene Sequence ◽  
Rrna Gene ◽  
Rrna Gene Sequence ◽  
Gene Sequence Analysis ◽  
Novel Genus And Species

A taxonomic study was carried out on a bacterial strain, designated KM-45T, isolated from forest soil collected near Daejeon, South Korea. Comparative 16S rRNA gene sequence analysis indicated a clear affiliation of this bacterium to the ‘Betaproteobacteria’ and that it was related most closely to Chitinibacter tainanensis BCRC 17254T, Formivibrio citricus DSM 6150T and Iodobacter fluviatilis ATCC 33051T (92·4, 91·2 and 88·9 % 16S rRNA gene sequence similarity, respectively). Cells were Gram-negative, facultatively anaerobic, motile and rod-shaped. The strain grew well on R2A medium and utilized a broad spectrum of carbon sources. The G+C content of the genomic DNA was 58 mol% and the predominant ubiquinone was Q-8. Major fatty acids were C16 : 0, C16 : 1 ω7c/iso-C15 : 0 2-OH, C18 : 1 ω7c/ω9t/ω12t and C17 : 0 cyclo. On the basis of the evidence presented, it is proposed that strain KM-45T should be placed in a novel genus and species, for which the name Silvimonas terrae gen. nov., sp. nov. is proposed. The type strain is KM-45T (=KCTC 12358T=NBRC 100961T).

scholarly journals Desulfurispirillum indicum sp. nov., a selenate- and selenite-respiring bacterium isolated from an estuarine canal

2011 ◽  
Vol 61 (3) ◽  
pp. 654-658 ◽  
Author(s):  
Ines Rauschenbach ◽  
Priya Narasingarao ◽  
Max M. Häggblom
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Carbon Sources ◽  
Cellular Fatty Acid ◽  
Elemental Selenium ◽  
Rrna Gene ◽  
Novel Bacterium ◽  
The Family ◽  
Type Strains ◽  
Fatty Acid Compositions

Strain S5T, a novel bacterium that was isolated for its capability to respire selenate to elemental selenium, is described. In addition to selenate respiration, it was also capable of dissimilatory selenite, arsenate and nitrate reduction with short-chain organic acids such as pyruvate, lactate and acetate as the carbon sources and electron donors. The isolate was unable to grow fermentatively. Strain S5T was isolated from sediment of an estuarine canal in Chennai, India. Phylogenetic analysis of the 16S rRNA gene of this novel isolate revealed that it belonged to the family Chrysiogenaceae with sequence similarities of 92 and 98 %, respectively, with the type strains of Chrysiogenes arsenatis and Desulfurispirillum alkaliphilum, its closest known relatives. Strain S5T and D. alkaliphilum were closely related in terms of their 16S rRNA gene phylogeny; however, they varied greatly in their genomic DNA G+C content (56 mol% versus 45 mol%) and cellular fatty acid compositions, as well as in many metabolic capabilities. Strain S5T represents a novel species for which the name Desulfurispirillum indicum sp. nov. is proposed; the type strain is S5T (=DSM 22839T =ATCC BAA-1389T).

scholarly journals Kordiimonas gwangyangensis gen. nov., sp. nov., a marine bacterium isolated from marine sediments that forms a distinct phyletic lineage (Kordiimonadales ord. nov.) in the ‘Alphaproteobacteria’

2005 ◽  
Vol 55 (5) ◽  
pp. 2033-2037 ◽  
Author(s):  
Kae Kyoung Kwon ◽  
Hee-Soon Lee ◽  
Sung Hyun Yang ◽  
Sang-Jin Kim
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Marine Bacterium ◽  
Enrichment Culture ◽  
Sequence Similarity ◽  
Bacterial Species ◽  
Carbon Sources ◽  
Optimal Growth ◽  
Rrna Gene ◽  
Respiratory Quinone

A marine bacterium, designated strain GW14-5T, capable of degrading high-molecular-mass polycyclic aromatic hydrocarbons was isolated from the sediments of Gwangyang Bay, Republic of Korea, after enrichment culture for 2 years with a mixture of benzo[a]pyrene and pyrene. Phylogenetic analysis based on 16S rRNA gene sequences indicated that the isolate forms a phyletic lineage that is distinct from the seven known orders within the ‘Alphaproteobacteria’. 16S rRNA gene sequence similarity of strain GW14-5T to all recognized bacterial species was not greater than 92 %. The dominant fatty acids of the isolate were i-17 : 1 (46·2 %), i-15 : 0 (15·1 %) and i-17 : 0 (12·6 %). The major respiratory quinone was MK-5, and the DNA G+C content was 39·3 mol%. Cells of strain GW14-5T were Gram-negative, motile, catalase-positive, oxidase-positive and weakly halophilic. Glucose, N-acetylglucosamine and maltose were utilized as sole carbon sources. The strain was positive for β-glucosidase activity. Optimal growth of strain GW14-5T was at pH 7·0 and 37–40 °C and required the presence of 2 % (w/v) NaCl. On the basis of this evidence, strain GW14-5T represents a novel genus and species in the ‘Alphaproteobacteria’ for which the name Kordiimonas gwangyangensis gen. nov., sp. nov. is proposed. The novel order Kordiimonadales is proposed for the distinct phyletic line represented by the genus Kordiimonas. The type strain is GW14-5T (=KCCM 42021T=JCM 12864T).

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