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1.
Int J Syst Evol Microbiol ; 69(10): 3128-3134, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31364963

RESUMO

A novel Gram-stain-positive actinobacterial strain, designated C9-28T, was isolated from soil sampled in a natural cave on Jeju Island, Republic of Korea. Strain C9-28T morphologically exhibited a rod-coccus life cycle and grew at 10-37 °C (optimum, 30 °C), pH 6-9 (optimum, pH 7) and 0-3 % (optimum, absence of NaCl). In the maximum-likelihood tree based on 16S rRNA gene sequences, strain C9-28T formed a sublineage between a Rhodococcus equi-Rhodococcus soli-Rhodococcus agglutinans clade and the type strain of Rhodococcus defluvii. The closest relatives of strain C9-28T were the type strains of R. defluvii (98.88 % 16S rRNA gene sequence similarity), R. equi (98.88 %) and R. soli (98.60 %). The phylogenomic tree based on whole genome sequences supported the distinct position of the novel strain within the genus Rhodococcus. The following chemotaxonomic characteristics also supported the assignment to the genus: meso-diaminopimelic acid; arabinose and galactose in whole-cell hydrolysates; the predominant menaquinone of MK-8(H2); and polar lipids including diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylinositol, phosphatidylinositol mannoside, three unidentified glycolipids and two unidentified lipids. The predominant cellular fatty acids were C16 : 0, summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c), C18 : 1ω9c and C14 : 0. Based on the values of average nucleotide identity and digital DNA-DNA hybridization from whole genome sequences, and in vitro DNA-DNA hybridization between the isolate and the closest relatives, strain C9-28T (=KACC 19823T=DSM 107559T) represents a novel species of the genus Rhodococcus, for which the name Rhodococcussubtropicus sp. nov. is proposed.


Assuntos
Cavernas/microbiologia , Filogenia , Rhodococcus/classificação , Microbiologia do Solo , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/genética , Ácido Diaminopimélico/química , Ácidos Graxos/química , Hibridização de Ácido Nucleico , Fosfolipídeos/química , RNA Ribossômico 16S/genética , República da Coreia , Rhodococcus/isolamento & purificação , Análise de Sequência de DNA , Vitamina K 2/análogos & derivados , Vitamina K 2/química
2.
Int J Syst Evol Microbiol ; 69(10): 2966-2971, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31418670

RESUMO

A novel actinobacterium, designated LHW52908T, was isolated from a marine sponge, Leucettachagosensis, collected in the South China Sea. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain LHW52908T was member of the family Geodermatophilaceae, with highest similarities to Geodermatophilus obscurus DSM 43160T (97.7 %), Geodermatophilus siccatus CF6T (97.6 %) and Geodermatophiluschilensis B12T (97.5 %). Multilocus sequence analysis confirmed that the strain should be a member of genus Geodermatophilus. Chemotaxonomic characteristics confirmed the genus-level affiliation of strain LHW52908T. Based on phylogenetic data, average nucleotide identity and digital DNA-DNA hybridization results, strain LHW52908T could be distinguished from its closest neighbours, representing a novel species of the genus Geodermatophilus, for which the name Geodermatophilusmarinus sp. nov. is proposed, with the type strain LHW52908T (=DSM 106570T=CCTCC AA 2018014T).


Assuntos
Actinobacteria/classificação , Filogenia , Poríferos/microbiologia , Actinobacteria/isolamento & purificação , Animais , Técnicas de Tipagem Bacteriana , Composição de Bases , China , DNA Bacteriano/genética , Ácidos Graxos/química , Tipagem de Sequências Multilocus , Hibridização de Ácido Nucleico , Oceanos e Mares , Fosfolipídeos/química , Pigmentação , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Vitamina K 2/análogos & derivados , Vitamina K 2/química
3.
Chem Commun (Camb) ; 55(69): 10288-10291, 2019 Aug 22.
Artigo em Inglês | MEDLINE | ID: mdl-31396601

RESUMO

A simple nanopore modification and sensing strategy was developed for the detection of miRNAs. This preparation and sensing approach provides a quick, simple and facile tool for the detection of specific biomolecules with high sensitivity and selectivity, and may find a wide range of applications in bio-analysis.


Assuntos
Ouro/química , Ácidos Nucleicos Imobilizados/química , MicroRNAs/análise , Nanoporos/ultraestrutura , Técnicas Biossensoriais/instrumentação , Desenho de Equipamento , Hibridização de Ácido Nucleico
4.
Chem Commun (Camb) ; 55(69): 10300-10303, 2019 Aug 22.
Artigo em Inglês | MEDLINE | ID: mdl-31397452

RESUMO

Shorter DNA probes provide better specificity for hybridization, but they may not form stable duplexes at room temperature. In this study, we used thiazole orange to follow DNA hybridization upon freezing and achieved stable 5-mer duplex DNA. Using multiple short probes in tandem, long DNA could also be studied. This study provides insights into DNA hybridization in the frozen state and expands the application of freezing for nucleic acid chemistry.


Assuntos
DNA/química , Hibridização de Ácido Nucleico , Oligonucleotídeos/química , Pareamento Incorreto de Bases , Sequência de Bases , Benzotiazóis/análise , DNA/genética , Sondas de DNA/química , Sondas de DNA/genética , Corantes Fluorescentes/análise , Congelamento , Oligonucleotídeos/genética , Quinolinas/análise , Espectrometria de Fluorescência
5.
Int J Syst Evol Microbiol ; 69(10): 3022-3030, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31364965

RESUMO

Two novel strains, designated YLB-02T and YLB-04T, were isolated from the deep-sea sediments of Yap Trench located in the Pacific Ocean. Cells of the strains were Gram-stain-positive, oxidase- and catalase-positive and rod-shaped. Phylogenetic analyses based on 16S rRNA gene sequences indicated that strain YLB-02T belonged to the genus Oceanobacillus and strain YLB-04T belonged to the genus Bacillus. Strain YLB-02T showed similarities of 96.9 % with Ornithinibacillus contaminans CCUG 53201T, 96.3 % with Oceanobacillus profundus CL-MP28T, 96.1 % with Oceanobacillus halophilus J8BT and 95.7 % with Oceanobacillus bengalensis Ma-21T. Strain YLB-04T showed the highest sequence similarity of 97.4 % with Bacillus notoginsengisoli SYP-B691T. The average nucleotide identity (ANI) and the DNA-DNA hybridisation (DDH) estimate values for strain YLB-02T and YLB-04T with their related type strains were below the respective threshold for species differentiation. The G+C contents of strains YLB-02T and YLB-04T were 37.3 and 45.4 mol%. The predominant (>10 %) cellular fatty acids of strain YLB-02T were iso-C14 : 0, iso-C15 : 0, iso-C16 : 0 and C16 : 1ω7c alcohol, and those of strain YLB-04T were C16 : 0, iso-C15 : 0, anteiso-C15 : 0 and C18 : 0. Their predominant ubiquinone was MK-7. The cell-wall peptidoglycan of strain YLB-02T contained glutamic acid, alanine, aspartic acid, lysine and ornithine, but no meso-diaminopimelic acid, while strain YLB-04T contained meso-diaminopimelic acid, glutamic acid, alanine, aspartic acid, lysine and ornithine. In addition to diphosphatidylglycerol (DPG) and phosphatidylglycerol (PG), the polar lipids of strain YLB-02T also consisted of an unidentified glycolipid (GL), two unidentified polar lipids (L1 and L2) and two unidentified phospholipids (PL1 and PL2), and those of strain YLB-04T also consisted of phosphatidylethanolamine (PE) and an unidentified phospholipid (PL). Based on phenotypic, genotypic and chemotaxonomic characteristics, two novel species are proposed, Oceanobacillus piezotolerans sp. nov. with YLB-02T (=MCCC 1A12699T=JCM 32870T) and Bacillus piezotolerans sp. nov. with YLB-04T (=MCCC 1A12711T=JCM 32872T) as the type strains.


Assuntos
Bacillaceae/classificação , Bacillus/classificação , Sedimentos Geológicos/microbiologia , Filogenia , Água do Mar/microbiologia , Bacillaceae/isolamento & purificação , Bacillus/isolamento & purificação , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/genética , Ácido Diaminopimélico/química , Ácidos Graxos/química , Hibridização de Ácido Nucleico , Oceano Pacífico , Peptidoglicano/química , Fosfolipídeos/química , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Vitamina K 2/análogos & derivados , Vitamina K 2/química
6.
Int J Syst Evol Microbiol ; 69(10): 3293-3298, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31385783

RESUMO

A Gram-negative, rod-shaped, non-motile and strictly aerobic bacterium, designated ZQ420T, was isolated from marine sediment sampled on Zhoushan Island located in the East China Sea. Strain ZQ420T was able to grow at 10-45 °C, 0-12.0 % (w/v) NaCl and pH 5.5-9.0. Catalase and oxidase activities, nitrate reduction, H2S production, hydrolysis of starch, casein, Tween 20, 40 and 80 were positive. Indole, methyl red, Voges-Proskauer test, hydrolysis of gelatin and Tween 60 were negative. The major cellular fatty acids were C18 : 1 ω7c, C16 : 0 and 11-methyl C18 : 1ω7c. Ubiquinone-10 (Q-10) was the only detected respiratory quinone. The polar lipids consisted of diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, three unidentified phospholipids, three unidentified glycolipids, two unidentified aminolipid and two unidentified lipids. The DNA G+C content was 64.8 mol%. According to 16S rRNA gene sequence similarities, strain ZQ420T shared 97.9, 96.3 and 96.3 % similarities to the following species with validated names Pararhodobacteraggregans D1-19T, Pseudo rhodobacter psychrotolerans PAMC27389T and Pseudo rhodobacter collinsensis 4-T-34T, respectively. While sharing lower sequence similarities (<96.0 %) to other type species. Phylogenetic analyses showed that strain ZQ420T and P. aggregans D1-19T formed an independent cluster in the phylogenetic trees. The average nucleotide identity value between strain ZQ420T and P. aggregans D1-19T was 79.1 %. The in silico DNA-DNA hybridization analysis revealed that strain ZQ420T shared 21.5 % DNA relatedness with P. aggregans D1-19T. On the basis of its phenotypic, chemotaxonomic and genotypic characteristics, strain ZQ420T is considered to represent a novel species in the genus Pararhodobacter, for which the name Pararhodobactermarinus sp. nov. is proposed. The type strain is ZQ420T (=KCTC 62579T=MCCC 1K03530T).


Assuntos
Sedimentos Geológicos/microbiologia , Filogenia , Rhodobacteraceae/classificação , Água do Mar/microbiologia , Técnicas de Tipagem Bacteriana , Composição de Bases , China , DNA Bacteriano/genética , Ácidos Graxos/química , Glicolipídeos/química , Hibridização de Ácido Nucleico , Fosfolipídeos/química , RNA Ribossômico 16S/genética , Rhodobacteraceae/isolamento & purificação , Análise de Sequência de DNA , Ubiquinona/química
7.
Int J Syst Evol Microbiol ; 69(10): 3299-3304, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31395107

RESUMO

A halophilic archaeaon, strain LT12T, was isolated from saline soil sampled at the Tarim Basin, PR China. The novel strain stained Gram-negative, cells were rod-shaped, and formed light red-pigmented colonies on agar plate. Strain LT12T grew optimally at 3.1 M NaCl, 0.05 M MgCl2, 37 °C and pH 7.5. The cells lysed in distilled water and the minimal NaCl concentration to prevent cell lysis was 1.4 M. Based on the results of phylogenetic analyses of the 16S rRNA and rpoB' genes, strain LT12T was most closely related to Halostella salina CBA1114T(94.4-95.9  and 93.6 % similarities, respectively). The average nucleotide identity and in silico DNA-DNA hybridization values between strain LT12T and H. salina CBA1114T were 81.0 and 24.3 %, respectively. The major polar lipids of strain LT12T were phosphatidic acid, phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester and three unidentified glycolipids. The DNA G+C content was 67.2 mol % (genome). Based on the phenotypic, chemotaxonomic and phylogenetic properties, strain LT12T represents a novel species of the genus Halostella for which the name Halostellalimicola sp. nov. is proposed. The type strain is LT12T (=CGMCC 1.14941T=JCM 30667T).


Assuntos
Halobacteriaceae/classificação , Filogenia , Salinidade , Microbiologia do Solo , Técnicas de Tipagem Bacteriana , Composição de Bases , China , DNA Arqueal/genética , DNA Bacteriano/genética , Ácidos Graxos/química , Genes Bacterianos , Glicolipídeos/química , Halobacteriaceae/isolamento & purificação , Hibridização de Ácido Nucleico , Fosfolipídeos/química , Pigmentação , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
8.
Int J Syst Evol Microbiol ; 69(10): 3093-3099, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31329533

RESUMO

A novel bacterial strain, designated NEAU-SA2T, was isolated from forest soil collected from the Zhangjiajie city, Hunan Province, PR China and characterised using a polyphasic approach. The cells were aerobic, Gram-stain-positive, non-flagellated and rod-coccus-shaped. The strain grew optimally at 28 °C, pH 7.0 and with 0 % NaCl (w/v). Phylogenetic analysis based on the 16S rRNA gene sequence indicated that the organism should be assigned to the genus Arthrobacter and was closely related to Arthrobacter cupressi DSM 24664T (98.89 %) and Arthrobacter silvisoli CCTCC AB 2017271T (98.41 %), which was further confirmed by multilocus sequence analysis. The major cellular fatty acids were anteiso-C15 : 0, anteiso-C17 : 0 and C16 : 0; MK-9(H2) was the predominant respiratory quinone. The polar lipids comprised diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol and an unidentified glycolipid. The peptidoglycan type was A3α, and the cell-wall sugars were glucose and galactose. The genomic G+C content of strain NEAU-SA2T was 67.04 mol%. The average nucleotide identity values between NEAU-SA2T and A. cupressi DSM 24664T and A. silvisoli CCTCC AB 2017271T were 88.57-90.94 %. The digital DNA-DNA hybridisation values between strain NEAU-SA2T and its most closely related species were 37.00 and 41.10 %, respectively, again indicating that they belong to different taxa. Therefore, strain NEAU-SA2T represents a novel species of the genus Arthrobacter, for which the name Arthrobacter celericrescens sp. nov. is proposed. The type strain is NEAU-SA2T (=DSM 106718T=CCTCC AB 2017272T).


Assuntos
Arthrobacter/classificação , Florestas , Filogenia , Microbiologia do Solo , Arthrobacter/isolamento & purificação , Técnicas de Tipagem Bacteriana , Composição de Bases , Parede Celular/química , China , DNA Bacteriano/genética , Ácidos Graxos/química , Glicolipídeos/química , Hibridização de Ácido Nucleico , Peptidoglicano/química , Fosfolipídeos/química , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Vitamina K 2/análogos & derivados , Vitamina K 2/química
9.
Int J Syst Evol Microbiol ; 69(10): 3135-3140, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31334696

RESUMO

A Gram-staining-negative, aerobic, motile with a single polar flagellum and rod-shaped bacterium as a bacterial host of podovirus P26218, designated IMCC26218T, was isolated from Lake Soyang, South Korea. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain IMCC26218T belonged to the genus Rhodoferax of the family Comamonadaceae and shared 97.7-99.0 % sequence similarities with Rhodoferax species. The draft whole genome sequence of strain IMCC26218T was ca. 4.9 Mbp in size with the DNA G+C content of 62.3 mol%. Average nucleotide identity (ANI) and digital DNA-DNA hybridisation (dDDH) values between strain IMCC26218T and other Rhodoferax were 74.0-77.3 % and 19.5-21.0 %, respectively, showing that the strain represents a new Rhodoferax species. The strain contained summed feature 3 (C16 : 1 ω6c and/or C16 : 1 ω7c) and C16 : 0 as the major fatty acids and phosphatidylethanolamine, three unidentified phospholipids, two unidentified aminolipids and two unidentified lipids as major polar lipids. The predominant isoprenoid quinone of the strain was ubiquinone-8 (Q-8). On the basis of the phylogenetic and phenotypic characteristics, strain IMCC26218T is considered to represent a novel species of the genus Rhodoferax, for which the name Rhodoferax lacus sp. nov. is proposed. The type strain is IMCC26218T (=KACC 18983T=NBRC 112709T).


Assuntos
Comamonadaceae/classificação , Lagos/microbiologia , Filogenia , Técnicas de Tipagem Bacteriana , Composição de Bases , Comamonadaceae/isolamento & purificação , DNA Bacteriano/genética , Ácidos Graxos/química , Hibridização de Ácido Nucleico , Fosfolipídeos/química , RNA Ribossômico 16S/genética , República da Coreia , Análise de Sequência de DNA , Ubiquinona/química
10.
Int J Syst Evol Microbiol ; 69(10): 3178-3190, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31339481

RESUMO

Fourteen Gram-stain-positive bacterial strains were isolated from Chinese traditional pickle and yogurt. The strains were characterised using a polyphasic taxonomic approach, including 16S rRNA gene sequence analysis, pheS gene sequence analysis, rpoA gene sequence analysis, determination of DNA G+C content, determination of average nucleotide identity, fatty acid methyl ester analysis and an analysis of phenotypic features. The data demonstrated that the fourteen strains represented nine novel species belonging to the genus Lactobacillus, strains 54-2T, 54-5T, 33-7T, 116-2T, 184-8T, 204-8T, 8-1(1)T, 256-3T and M1575T were designated as the type strains. Strain 54-2T was phylogenetically related to the type strains of Lactobacillus composti and Lactobacillus floricola, having 96.5 and 91.6 % 16S rRNA gene sequence similarities, less than 74.6 % pheS gene sequence similarities, less than 81.6 % rpoA gene sequence similarities and less than 72.5 % ANI values. Strain 54-5T was phylogenetically related to the type strains of Lactobacillus dextrinicus and Lactobacillus concavus, exhibiting 99.1 and 97.3 % 16S rRNA gene sequence similarities, less than 83.1 % pheS gene sequence similarities, less than 93.1 % rpoA gene sequence similarities and less than 79.9 % ANI values. Strains 33-7T, 116-2T, 184-8T, 204-8T, 8-1(1)T, 256-3T and M1575T were phylogenetically related to the type strains of Lactobacillus tucceti, Lactobacillus nodensis, Lactobacillus insicii, Lactobacillus allii, Lactobacillus metriopterae, Lactobacillus terrae, Lactobacillus versmoldensis and Lactobacillus furfuricola, sharing 95.6-100 % 16S rRNA gene sequence similarities, less than 91.6 % pheS gene sequence similarities, less than 98.2 % rpoA gene sequence similarities and less than 89.4 % ANI values. Based upon the data of polyphasic characterisation obtained in the present study, nine novel species, Lactobacillus yilanensis sp. nov., Lactobacillus bayanensis sp. nov., Lactobacillus keshanensis sp. nov., Lactobacillus kedongensis sp. nov., Lactobacillus baiquanensis sp. nov., Lactobacillus jidongensis sp. nov., Lactobacillus hulinensis sp. nov., Lactobacillus mishanensis sp. nov. and Lactobacillus zhongbaensis sp. nov., are proposed and the type strains are 54-2T (=NCIMB 15154T=CCM 8896T=KCTC 21120T=LMG 31058T), 54-5T (=NCIMB 15151T=CCM 8894T), 33-7T (=NCIMB 15153T=CCM 8936T=KCTC 21118T=LMG 31166T), 116-2T (=NCIMB 15158T=CCM 8899T=KCTC 21124T=LMG 31051T), 184-8T (=NCIMB 15152T=CCM 8895T=KCTC 21131T=LMG 31050T), 204-8T (=NCIMB 15159T=CCM 8900T=KCTC 21133T=LMG 31054T), 8-1(1)T (=NCIMB 15156T=CCM 8898T=KCTC 21115T=LMG 31047T), 256-3T (=NCIMB 15160T=CCM 8901T=LMG 31048T) and M1575T (=NCIMB 15149T=CCM 8892T=LMG 31045T), respectively.


Assuntos
Alimentos Fermentados/microbiologia , Microbiologia de Alimentos , Lactobacillus/classificação , Filogenia , Iogurte/microbiologia , Técnicas de Tipagem Bacteriana , Composição de Bases , China , DNA Bacteriano/genética , Ácidos Graxos/química , Genes Bacterianos , Lactobacillus/isolamento & purificação , Hibridização de Ácido Nucleico , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
11.
Int J Syst Evol Microbiol ; 69(10): 3191-3201, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31339483

RESUMO

Five novel lactic acid bacterial strains, isolated from Chinese traditional pickle juice, were characterised by a polyphasic approach, including 16S rRNA gene sequence analysis, pheS gene sequence analysis, rpoA gene sequence analysis, determination of DNA G+C content, average nucleotide identity (ANI) analysis, in silico DNA-DNA hybridisation (isDDH), fatty acid methyl ester (FAME) analysis and an analysis of phenotypic features. Strains 241-2-2T, 63-4T and 190-7T were closely related to the type strains of Enterococcus devriesei, Enterococcus viikkiensis, Enterococcus pseudoavium, Enterococcus xiangfangensis, Enterococcus avium, Enterococcus malodoratus, Enterococcus raffinosus and Enterococcus gilvus, having 99.1-99.9 % 16S rRNA gene sequence similarities. Strain 94-2T was distantly related to the type strains of Enterococcus phoeniculicola, Enterococcus rivorum and Enterococcus faecalis, having 95.2-96.1 % 16S rRNA gene sequence similarities. Strain 85-4T was distantly related to the type strains of Enterococcus casseliflavus, Enterococcus gallinarum, Enterococcus dispar, Enterococcus canintestini, Enterococcus saigonensis, Enterococcus diestrammenae, Enterococcus asini, Enterococcus cecorum and Enterococcus columbae, having 95.7-97.8 % 16S rRNA gene sequence similarities. Less than 91.5 % ANI and 45.3 % isDDH values between strains 241-2-2T, 63-4T, 190-7T, 94-2T, 85-4T and type strains of phylogenetically related species showed that they represent five new species within the genus Enterococcus. Based upon the data of polyphasic characterisation obtained in the present study, five novel species, Enterococcus pingfangensis sp. nov., Enterococcus dongliensis sp. nov., Enterococcus hulanensis sp. nov., Enterococcus nangangensis sp. nov. and Enterococcus songbeiensis sp. nov., are proposed and the type strains are 241-2-2T (=NCIMB 15185T=CCM 8921T=LMG 31181T), 63-4T (=NCIMB 15178T=CCM 8922T), 190-7T (=NCIMB 15200T=CCM 8949T), 94-2T (=NCIMB 15180T=CCM 8920T=LMG 31180T) and 85-4T (=NCIMB 15179T=CCM 8923T=LMG 31183T), respectively.


Assuntos
Enterococcus/classificação , Alimentos Fermentados/microbiologia , Microbiologia de Alimentos , Filogenia , Técnicas de Tipagem Bacteriana , Composição de Bases , China , DNA Bacteriano/genética , Enterococcus/isolamento & purificação , Ácidos Graxos/química , Genes Bacterianos , Hibridização de Ácido Nucleico , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
12.
Int J Syst Evol Microbiol ; 69(10): 3207-3216, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31343401

RESUMO

The Pectobacteriumcarotovorum species corresponds to a complex, including two subspecies with validly published names, two proposed subspecies and two new species, Pectobacterium polaris and Pectobacterium aquaticum. Recent studies suggested that this complex needed revision. We examined the taxonomic status of 144 Pectobacterium strains isolated from a wide range of plant species, various geographical origins and waterways. Sequences of the leuS, dnaX and recA housekeeping genes clustered 114 of these Pectobacterium strains together within a not yet described clade. We sequenced eight strains of this clade and analysed them together with the 102 Pectobacterium genomes available in the NCBI database. Phylogenetic analysis, average nucleotide identity calculation and in silico DNA-DNA hybridization allowed us to differentiate seven clades. This led us to propose the elevation of Pectobacterium carotovorumsubsp. odoriferum to species level as Pectobacteriumodoriferum sp. nov. (type strain CFBP 1878T=LMG 5863T=NCPPB 3839T=ICMP 11533T), the proposal of Pectobacteriumactinidiae sp. nov. (type strain KKH3=LMG 26003 T=KCTC 23131T) and Pectobacteriumbrasiliense sp. nov. (type strain CFBP 6617T= LMG 21371T=NCPPB 4609T), to emend the description of Pectobacterium carotovorum (type strain CFBP 2046T=LMG 2404T=NCPPB 312T=ICMP 5702T), and to propose a novel species, Pectobacterium versatile sp. nov (type strain CFBP6051T= NCPPB 3387T=ICMP 9168T) which includes the strains previously described as 'Candidatus Pectobacterium maceratum'. Phenotypic analysis performed using Biolog GENIII plates on eight strains of P. versatile sp. nov. and related strains completed our analysis.


Assuntos
Pectobacterium carotovorum/classificação , Pectobacterium/classificação , Filogenia , Plantas/microbiologia , Rios/microbiologia , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/genética , Ácidos Graxos/química , França , Genes Bacterianos , Líbano , Marrocos , Hibridização de Ácido Nucleico , Pectobacterium/isolamento & purificação , Pectobacterium carotovorum/isolamento & purificação , Doenças das Plantas/microbiologia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
13.
Int J Syst Evol Microbiol ; 69(10): 3155-3160, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31355737

RESUMO

A novel Kosakonia strain WCHEs120001T was recovered from the wound secretion of a patient at West China Hospital, Chengdu, PR China, in 2017. The strain was Gram-stain-negative, facultatively anaerobic, motile, and non-spore-forming. A preliminary analysis based on the 16S rRNA gene sequence revealed that this strain was closely related to members of the genus Kosakonia. The strain was subjected to whole genome sequencing. Phylogenetic analysis based on core gene sequences of type strains of all Enterobacteriaceae species revealed that this strain belonged to the genus Kosakonia but were distinct from any previously known Kosakonia species. Both average nucleotide identity (ANI) and in silico DNA-DNA hybridisation (isDDH) values between strain WCHEs120001T and type strains of all known Kosakonia species were 82.02 to 92.37% and 25.6 to 50.9 %, respectively, which are lower than the 95 % (ANI) and 70 % (isDDH) cutoff for species delineation. The major fatty acids of the strain WCHEs120001T are C16 : 0, sum of C16:1ω7c/C16:1ω6c and C18:1ω7c, which are similar to other Kosakonia species. Genomic DNA G+C content of strain WCHEs120001T was 53.33 mol%. Strain WCHEs120001T is positive for methyl-d-glucopyranoside but does not ferment adonitol, d-arabitol, dulcitol and melibiose, which distinguishes it from all other Kosakonia species. Genotypic and phenotypic characteristics indicate that strain WCHEs120001T represents a novel species of the genus Kosakonia, for which the name Kosakonia quasisacchari sp. nov. is proposed. The type strain of K. quasisacchari sp. nov. is WCHEs120001T (=GDMCC1.1570T=NCTC 14272T).


Assuntos
Enterobacteriaceae/classificação , Filogenia , Infecção dos Ferimentos/microbiologia , Técnicas de Tipagem Bacteriana , Composição de Bases , China , DNA Bacteriano/genética , Enterobacteriaceae/isolamento & purificação , Ácidos Graxos/química , Humanos , Hibridização de Ácido Nucleico , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
14.
Int J Syst Evol Microbiol ; 69(10): 3237-3247, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31361212

RESUMO

Seven novel lactic acid bacterial strains, isolated from traditional Chinese pickle, were characterized using a polyphasic approach, including 16S rRNA gene sequence analysis, pheS gene sequence analysis, rpoA gene sequence analysis, determination of DNA G+C content, determination of average nucleotide identity (ANI), in silico DNA-DNA hybridization (isDDH), fatty acid methyl ester (FAME) analysis and an analysis of phenotypic features. Strains 382-1T, 116-1AT, 381-7T, 203-3T, 218-3T and 398-2T were phylogenetically related to the type strains of Lactobacillus plantarumsubsp. plantarum, Lactobacillus plantarumsubsp. argentoratensis, Lactobacillus pentosus, Lactobacillus paraplantarum, Lactobacillus fabifermentans, Lactobacillus herbarum, Lactobacillus mudanjiangensis, Lactobacillus xiangfangensis, Lactobacillus plajomi and Lactobacillus modestisalitolerans, having 97.1-99.9 % 16S rRNA gene sequence similarities, less than 89.9 % pheS gene sequence similarities, less than 98.0 % rpoA gene sequence similarities, less than 91.2 % ANI values and less than 43.3 % isDDH values. Strain 778-3T was phylogenetically related to the type strains of Lactobacillus hokkaidonensis, Lactobacillus wasatchensis, Lactobacillus oligofermentans, Lactobacillus nenjiangensis, Lactobacillus vaccinostercus and Lactobacillus suebicus, exhibiting 97.0-99.4 % 16S rRNA gene sequence similarities, 78.2-82.1 % pheS gene sequence similarities, 80.0-91.5 % rpoA gene sequence similarities, less than 78.6 % ANI values and less than 22.9 % isDDH values. Based upon the data of polyphasic characterization obtained in the present study, seven novel species, Lactobacillus pingfangensis sp. nov., Lactobacillus daoliensis sp. nov., Lactobacillus nangangensis sp. nov., Lactobacillus daowaiensis sp. nov., Lactobacillus dongliensis sp. nov., Lactobacillus songbeiensis sp. nov. and Lactobacillus kaifaensis sp. nov., are proposed and the type strains are 382-1T (=NCIMB 15187T=CCM 8935T=LMG 31176T), 116-1AT (=NCIMB 15181T=CCM 8934T=LMG 31171T), 381-7T (=NCIMB 15186T=CCM 8930T), 203-3T (=NCIMB 15183T=CCM 8933T=LMG 31172T), 218-3T (=NCIMB 15184T=CCM 8932T=LMG 31173T), 398-2T (=NCIMB 15189T=CCM 8931T=LMG 31174T) and 778-3T (=NCIMB 15191T=CCM 8929T=LMG 31177T), respectively.


Assuntos
Alimentos Fermentados/microbiologia , Microbiologia de Alimentos , Lactobacillus/classificação , Filogenia , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/genética , Ácidos Graxos/química , Genes Bacterianos , Lactobacillus/isolamento & purificação , Hibridização de Ácido Nucleico , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
15.
18.
Analyst ; 144(16): 4917-4924, 2019 Aug 21.
Artigo em Inglês | MEDLINE | ID: mdl-31313769

RESUMO

MicroRNAs (miRNAs) are attractive candidates for biomarkers for early cancer diagnosis, and play vital roles in physiological and pathological processes. In this work, we developed a colorimetric and fluorescent dual-mode sensor for miRNA detection based on the optical properties of gold nanoparticles (AuNPs) and the duplex-specific nuclease (DSN)-assisted signal amplification technique. In brief, FAM labelled hairpin probes (HPs) were immobilized on AuNPs, and fluorescence was efficiently quenched by the vicinity of the fluorophores to the AuNPs surface. In the presence of target miRNAs, the HPs could specifically hybridize with miRNAs and the DNA strand in the DNA/RNA heteroduplexes could be subsequently hydrolyzed by DSN. As a result, numbers of fluorophores were released into the solution, resulting in obvious fluorescence signal recovery. Meanwhile, the target miRNAs were able to participate in other hybridization reactions. With the DSN-assisted signal amplification technique, lots of gold nanoparticles were produced with short-chain DNA on their surface, which could aggregate in salt solution and result in a colorimetric detection. The proposed dual-mode strategy offers a sensitive, accurate and selective detection method for miRNAs. One reason is that the stem of the HPs was elaborately designed to avoid hydrolyzation by DSN under optimal conditions, which ensures a relatively low background and high sensitivity. The other is that the dual-mode strategy is more beneficial for enhancing the accuracy and reproducibility of the measurements. Moreover, the unique selective-cutting ability and single-base mismatch differentiation capability of the DSN also give rise to a satisfactory selectivity. This demonstrated that the developed method could quantitatively detect miR-21 down to 50 pM with a linear calibration range from 50 pM to 1 nM, and the analytical assay of target miRNAs in cell lysate samples revealed its great potential for application in biomedical research and clinical diagnostics.


Assuntos
Corantes/química , Endonucleases/química , Ouro/química , Nanopartículas Metálicas/química , MicroRNAs/análise , Técnicas Biossensoriais/métodos , Linhagem Celular , Colorimetria , DNA/química , Humanos , Limite de Detecção , Técnicas de Amplificação de Ácido Nucleico/métodos , Conformação de Ácido Nucleico , Ácidos Nucleicos Heteroduplexes/química , Hibridização de Ácido Nucleico , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Espectrometria de Fluorescência
19.
Chem Commun (Camb) ; 55(58): 8466-8469, 2019 Jul 25.
Artigo em Inglês | MEDLINE | ID: mdl-31265022

RESUMO

We presented a branch migration based PCR in which a branch migration blocker was introduced to selectively reduce the amplification efficiency of the wild-type target and enrich the mutant-type target. The low-abundance mutations could be enriched and then detected by high resolution melting, Sanger sequencing or fluorescent DNA probe.


Assuntos
Análise Mutacional de DNA/métodos , DNA/análise , DNA/genética , Reação em Cadeia da Polimerase/métodos , Sequência de Bases , Sondas de DNA/química , Sondas de DNA/genética , DNA Polimerase Dirigida por DNA/química , Fluorescência , Corantes Fluorescentes/química , Genes , Humanos , Limite de Detecção , Mutação , Hibridização de Ácido Nucleico
20.
Int J Syst Evol Microbiol ; 69(9): 2892-2898, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31259677

RESUMO

A Gram-stain-negative, non-flagellated, catalase-positive, oxidase-positive bacterial strain, designated YLY08T, was isolated from the gut microflora of sea bass (Dicentrarchus labrax L.) collected from the coast of Yuanyao Wharf, Weihai, PR China, and subjected to a polyphasic taxonomic study. Strain YLY08T grew optimally at 28-30 °C, at pH 7.0-7.5 and in the presence of 2.0-3.0 % (w/v) NaCl. Poly-ß-hydroxybutyrate granules were produced. Neighbour-joining, maximum-likelihood and maximum-parsimony phylogenetic trees based on 16S rRNA gene sequences revealed that strain YLY08T clustered with the type strain of Oceaniglobus indicus, with which it exhibited 95.3 % sequence similarity, while the similarity to other genera was below 95.0 %. Genomic analyses, including average nucleotide identity and the digital DNA-DNA hybridization, clearly separated YLY08T from O. indicus MCCC 1A11863T with values below the thresholds for species delineation. The major cellular fatty acid was summed feature 8 (C18 : 1ω7c and/or C18 : 1ω6c). The sole respiratory quinone detected was Q-10. The polar lipid profile consisted of phosphatidylethanolamine, phosphatidylglycerol, phosphatidylcholine, phosphatidyldimethylethanolamine, diphosphatidylglycerol, one unidentified aminolipid and one unidentified phospholipid. The genome of strain YLY08T, with 38 assembled contigs, was 3.9 Mb long with a G+C content of 59.0 mol%. The results of the phenotypical, phylogenetic and biochemical analyses between the strain YLY08T and the related type strain indicated that this strain represents a novel species in genus Oceaniglobus within the family Rhodobacteraceae, for which the name Oceaniglobus ichthyenteri sp. nov. is proposed. The type strain is YLY08T (=MCCC 1H00318T=KCTC 62182T).


Assuntos
Bass/microbiologia , Microbioma Gastrointestinal , Filogenia , Rhodobacteraceae/classificação , Animais , Técnicas de Tipagem Bacteriana , Composição de Bases , China , DNA Bacteriano/genética , Ácidos Graxos/química , Hidroxibutiratos , Hibridização de Ácido Nucleico , Fosfolipídeos/química , Poliésteres , RNA Ribossômico 16S/genética , Rhodobacteraceae/isolamento & purificação , Alimentos Marinhos/microbiologia , Água do Mar/microbiologia , Análise de Sequência de DNA
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