| The marine sediments of offshore have abundant microbial resources,and there are many potential functional bacteria and potential novel species,which have great exploitation potentialities.Based on the traditional enrichment culture method and uncultured sequencing technology,the microbial diversity and community structure and the succession of bacterial florain in the process of enrichment culture was studied in this paper.At the same time,fifty-six potential novel species were discovered.In addition,based on the polyphasic analysis,five strains were considered to represent novel species,and further enriched the bacterium resources.In this study,there were two kinds of enrichment culture method,named A and F.The traditional enrichment culture method and Illumina Miseq high-throughput sequencing technology found that the amount of bacteria was significantly increased compared to that of the non-enrichment culture,and the diversity was increased.With the extension of enriched time,the diversity of cultured bacteria increased firstly and then decreased,and the diversity was the highest at the 21st day.Based on the results,the enrichment culture method promoted the enrichment of Bacteroidetes,Firmicutes and Fusobacteria,but which was not conducive to the enrichment of Cyanobacteria,Meanwhile,compared with the A treatment,the abundance and diversity of bacteria were significantly reduced at the corresponding time in the F treatment,and the community structure was differential.According to this study,it could provide references for the isolation of specific bacterial and the improvement of enrichment culture method.Date in this study suggested that there were abundant microbial resources deserved to further development.Three novel marine strains were isolated from marine sediment sample collected from the coast of Weihai,designated FA042T,HF004T and D2T.Based on its phylogenetic and phenotypic characteristics,strain FA042T is presented as a representative of a novel species,for which the name Hyunsoonleella rubra sp.nov.is proposed;strain HF004T is presented as a representative of a novel species,for which the name Halioglobns lutimaris sp.nov.is proposed;strain D2rT is presented as a representative of a novel species,for which the name Catenovulum sediminis sp.nov.is proposed.A novel red-pigmented,aerobic,rod-shaped,non-flagellated and non-gliding bacterium,designated FA042T.was isolated from a marine sediment sample collected from the coast of Weihai,China.Optimal growth occurred at 33?,pH 7.0-7.5 and in the presence of 2-3%(w/v)NaCl.Phylogenetic analysis based on the 16S rRNA gene sequence showed that strain FA042T belonged to the genus Hyunsoonleella.The closest described neighbour,in terms of 16S rRNA gene sequence identity,was Hyunsoonleella jejuensis(95.0%).The major fatty acids were iso-C15:0,isO-C15;1 G,C15:0,iso-C17:0 3-OH and iso-C15:O 3-OH.The major respiratory quinone was MK-6.The major polar lipids of strain FA042T were phosphatidylethanolamine,three unidentified lipids and two unidentified aminolipids.The DNA G+C content was 38.5 mol%.Based on its phylogenetic and phenotypic characteristics,strain FA042T is presented as a representative of a novel species,for which the name Hyunsoonleella rubra sp.nov.is proposed.The type strain of Hyunsoonleella rubra sp.nov.is FA042T(=KCTC 42398T=MCCC 1H00110T).A novel Gram-stain-negative,aerobic,rod-shaped(approximately 1.3-1.8 ?m in length and 0.4-0.6 ?m in width),non-gliding,Gram-stain-negative bacterium,designated HF004T was isolated from a marine sediment sample collected from the coast of Weihai,China.Optimal growth occurred at 28?,pH 7.5-8.0 and in the presence of 2-3%(w/v)NaCl.Phylogenetic analysis based on 16S rRNA gene sequences revealed that HF004T had a 16S rRNA gene sequence similarity of 95.6-96.1%with recognized Halioglobus species.The major respiratory quinone was Q-8.The major fatty acids were summed feature 3(i.e.C16:1 ?7c and/or iso-C15:02-OH),C17;1 ?8c and C18:1 ?7c.The major polar lipids of strain HF004T were diphosphatidylglycerol,phosphatidylethanolamine and phosphatidylglycerol.The DNA G+C content was 56.9 mol%.On the basis of the phylogentic and phenotypic evidences,strain HF004T represents a novel species of the genus Halioglobus,for which the name Halioglobus lutimaris sp.nov.is proposed.The type strain is HF004-T(=KCTC 42395T=MCCC 1H00127T).A novel strictly aerobic,rod-shaped,agar-hydrolysing,motile by a lateral flagellun and Grain-stain-negative bacterium,designated D2L,was isolated from a marine sediment sample collected from the coast of Weihai,China.Growth was observed at 10-42?,at pH 6.0-9.0 and with 1-5%(w/v)NaCl.Phylogenetic analysis based on the 16S rRNA gene sequence showed that strain D2T belonged to the genus Catenovulum,appearing closely related to Catenovulum agarivorans YM01T(96.3%)and Catenovulum maritimus Q1T(93.9%).The dominant fatty acids were summed feature 3(i.e.C16:1?7c and/or iso-C15:02-OH),C16:0 and C10:0 3-OH.The major polar lipids were phosphatidylethanolamine,phosphatidylglycerol,one phospholipid and two phosphoaminolipids.Ubiquinone 8(Q-8)was found to be the major respiratory quinone.The DNA G+C content was determined to be 40.4 mol%.On the basis of genotypic,phenotypic and phylogenetic evidence,strain D2T is presented as a novel species,for which the name Catenovulum sediminis sp.nov.is proposed.The type strain of Catenovulum sediminis is D21(=KCTC 42869T=MCCC 1H00129T).Two novel strains were isolated from Gracilaria blodgettii,an algae of phylum Rhodophyta collected from coastal of Lingshui county,Hainan,China,designated 0182T and DC003T Based on its phylogenetic and phenotypic characteristics,strain 0182T is presented as a representative of a novel species,for which the name Crocinitomix algicola sp.nov.is proposed;strain DC003T is presented as a representative of a novel species,for which the name Reichenbachiella versicolor sp.nov.is proposed.A strictly aerobic,non-flagellated,yellow-pigmented,Gram-stain-negative,rod-shaped(approximately 0.9-2.5 ?m in length and 0.2-0.4 ?m in widtth)bacterial strain,designated 0182T,was isolated from Gracilaria blodgettii,an algae of phylum Rhodophyta collected from coastal of Lingshui county,Hainan,China.The strain grew optimally at 30?,pH 7.0-7.5 and in the presence of 2.0-3.0%(w/v)NaCl.Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain 0182T was phylogenetically related to members of the genus Crocinitomix and was closely related to Crocinitomix catalasitica with 94.6%sequence similarity.The major respiratory quinone was MK-7.The predominant cellular fatty acids were iso-Ci5:?,Ci5:0 and iso-C17;0 3-OH.The major polar lipids were phosphoaminolipid,two unknown aminolipids and four unknown lipids.The DNA G+C content was approximately 35.4 mol%.On the basis of genotypic and phenotypic characteristics and phylogenetic evidence,strain 0182-T is thought to represent a novel species of the genus Crocinitomix,for which the name Crocinitomix algicola sp.nov.is proposed.The type strain is 0182T(=KCTC 42868T=MCCC 1H00128T).A novel aerobic,non-flagellated,agar-hydrolysing and Gram-stain-negative bacterium,designated DC003T,was isolated from Gracilaria blodgettii,an algae of phylum Rhodophyta collected from coastal of Lingshui county,Hainan,China.Optimal growth occurred at 28?,pH 7.0-7.5 and in the presence of 2-3%(w/v)NaCl.Phylogenetic analysis based on 16S rRNA gene sequences revealed that DC003-T was a memeber of the family Flammeovirgaceae and had a 16S rRNA gene sequence similarity of 93.1-94.7%with recognized Reichenbachiella species.The major respiratory quinone was MK-7.The major fatty acids were iso-C15:0,summed feature 3(i.e.C16:1 ?7c and/or iso-C15:02-OH),C16:1 ?5c and summed feature 5(i.e.C18:a?6,9c and/or C18:0 ANTE).The major polar lipids of strain DC003-T were three unidentified lipids,two aminophospholipid,phosphatidyl ethanolamine and phospholipid.The DNA G+C content was 37.0 mol%.Based on its phylogenetic and phenotypic characteristics,strain DC003T is presented as a representative of a novel species,for which the name Reichenbachiella versicolor sp.nov.is proposed.The type strain of Reichenbachiella versicolor sp.nov.is DC003T(=KCTC 42867T=MCCC 1H00130T). |
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