Enrichment Culture And Isolation Of Bacteria From Coastal Sediment Of Weihai,Identification Of Novel Strains And Studies On The Predation Of B210

Diversity of microbe in offshore marine sediments is rich,containing a large number of potential novel bacteria resources.In this study,novel bacteria resources from offshore marine sediments were explored by enrichment culture method;the bacterial succession law of enrichment culture were detected by 454FLX+ platform.During the process of this study,amount of potential novel bacteria were found,7 novel marine strains were identified by polyphasic taxonomy.The predation phenomenon of novel bradymonabacteria B210 was firstly discovered,and the preliminarily exploration was carried on.326 strains were obtained by enrichment culture method,including 49 potential novel strains.The following conlusions could be made from metagemone sequencing analysis:there are the special bacteria flora during the different period of the enrichment,which could provide guides for the novel bactreia exploration;species aboundant declined as the time extend of enrichment culture until 30 days,it rised again;some rare bacteria increased by enrichment cultre,such as Acidobacteria.Novel agar-degrading bacterial strains Z1^T and JL1 were isolated from marine algae Gelidium amansii and Gracilaria verrucosa,respectively.Both strains contained Q-8 as the sole respiratory quinone.The major cellular fatty acids in strain Z1^T were C_18:1 ?7c,C_16:0 and summed feature 3(C_16:1?7c and/or iso-C_15:0 2-OH).The predominant polar lipids in strain Z1^T were phosphatidylethanolamine,phosphatidylglycerol and aminolipid.The genomic DNA G+C content of both strains was 45.1 mol%.Strains Z1^T and JLl shared the highest 16S rDNA sequences similarity to their closest relatives,with less than 92.3%.Based on data from the current polyphasic study,the isolates are proposed to belong to a new genus and species designated Marinagarivorans algicola gen.nov.,sp.nov.The type strain of Marinagarivorans algicola is Z1^T?=ATCC BAA-2617^T=CICC 10859^T?.Novel Gram-stain-variable strain NC2-42^T were isolated from the sediment on the coast of Weihai,China.MK-7 was the sole respiratory quinone and meso-diaminopimelic acid was a diagnostic diamino acid in the peptidoglycan.The polar lipid profile of this novel isolate consisted of phosphatidylglycerol,phosphatidylethanolamine,two unknown glycolipids%one unknown phospholipid,one unknown phosphoaminolipid and one unknown lipid.The major cellular fatty acids in strain NC2-42^T were anteiso-C_15:0,iso-C_16:0 and C_16:0.The G+C content of the genomic DNA of strain NC2-42^T was 58.1 mol%.Strain NC2-42^T showed the highest similarity?96.32%?to validly published taxon Paenibacillus profundus.Based on data from a current polyphasic study,the isolate was proposed to belong to a novel species of a new genus designated Marinicrinis sediminis gen.nov.,sp.nov.The type strain is NC2-42^T?=KCTC 33676^T=MCCC 1K01238^T?.A novel Gram-stain-negative strain XDB06^T was isolated from coastal sediment of Xiaoshi Island,Weihai,China.NaCl.Q-8 was the sole respiratory quinone.The major cellular fatty acids in strain XDB06^T were iso-C_15:0 and iso-C_16:0.The polar lipids of strain XDB06^T were diphosphatidylglycerol,phosphatidylglycerol,phosphatidylethanolamine,two unidentified glycolipids and four unidentified phospholipids.The genomic DNA G+C content of the strain was 65.0 mol%.Phylogenetic analysis based on 16S rDNA gene sequences showed that strain XDB06^T clusters within the genus Wenzhouxiangella and is most closely related to W.marina MCCC 1K00261^T,with a 16S rRNA gene sequence similarity of 96.5%.Based on data from the current polyphasic study,XDB06T represents a novel species within the genus Wenzhouxiangella,for which the name Wenzhouxiangella sediminis sp.nov.is proposed with type strain XDB06^T?=KCTC 52041^T= MCCC 1K02285^T?.A novel Gram-stain-positive strain 0W14^T,was isolated from marine saltern of Wendeng,China.MK7 was the sole respiratory quinone and the peptidoglycan type of OW14^T was A4? L-Orn-D-Glu.The major cellular fatty acid in strain 0W14^T was anteiso-C_15:0.The polar lipid profile of strain 0W14^T consisted of diphosphatidylglycerol,phosphatidylglycerol,two unknown glycolipids and four unknown phospholipids.The genomic DNA G+C content of the strain was 44.6 mol%.Strain 0W14^T shared the highest similarity?96.0%?to Virgibacillus litoralis.Based on data from the current polyphasic study,the isolate is proposed to represent a novel species of a new genus within the family Bacillaceae,with the name Marinisalinus sediminis gen.nov.,sp.nov.The type strain of the type species is 0W14^T?=KCTC 33835T=MCCC 1H00171^T?.A novel Gram-stain-positive strain NC2-31^T,was isolated from sediment on the coast of Weihai,China.MK7 was the major respiratory quinone and meso-diaminopimelic acid was a diagnostic diamino acid in the peptidoglycan.The major polar lipids of strain NC2-31^T were diphosphatidyl glycerol,phosphatidyl glycerol and phosphatidylethanolamine.The genomic DNA G+C content of the strain was 46.3 mol%.The predomi1ant cellular fatty acids in strain NC2-31^T were iso-C_15:0,anteiso-C_15:0,summed feature 3(C_16:1 ?7c and/or iso-C_15:02-OH)and iso-C_16:0.Phylogenetic analysis based on 16S rDNA sequences revealed that strain NC2-31^T formed a separated branch within the genus Bacillus.Based on data from the current polyphasic study,NC2-31^T represents a novel species within the genus Bacillus,for which the name Bacilus armisedimentorum sp.nov.is proposed with type strain NC2-31^T?=KCTC 33721^T= MCCC 1K01239^T?.A novel Gram-stain-negatiLe strain WDN1C137^T was isolated from marine saltern of Wendeng,China.Q-10 was the sole respiratory quinone.The major cellular fatty acids in strain WDN1C137^T were C_18:1?7c,cyclo C_19:0 ?8c and C_16:0.The major polar lipids are phosphatidylglycerol,phosphoglycolipid,one unknown glycolipid,one unknown lipid,two unknown aminolipids and three unknown phospholipids.The genomic DNA G+C was 71.4 mol%.Strain WDN1C137^T shared the highest similarity?94.5%?to Roseivivax jejudonensis.Based on data from the current polyphasic study,the isolate is proposed to represent a novel species of a new genus within the family Rhodobacteraceae,with the name Rhodosalinus sediminis gen.nov.,sp.nov.The type strain of the type species is WDN1C137^T?=KCTC 52478^T = MCCC 1H00170^T?.A novel Gram-stain-negative strain B210^T was isolated from the sediment on the coast of XisoShi island,Weihai,China.MK-7 was the sole respiratory quinone.The polar lipid profile of this novel isolate consisted of diphosphatidylglycerol,phosphatidylglycerol,phosphatidylethanolamine,one unknown phospholipidand and one unknown lipid.The major cellular fatty acids in strain B210^T was iso-C_15:0.The G+C content of the genomic DNA of strain B210^T was 64.7 mol%.A phylogenetic analysis based on the 16S rDNA gene sequences revealed that strain B210^T showed the highest similarity?89.2%?to Bradymonas sediminis FA350^T within the order Bradymonadales.Based on data from a current polyphasic study,the isolate was proposed to belong to a novel species of a new genus.The type strain is B210^T?-KCTC 42951^T?.The predation phenomenon of novel bradymonabacteria B210 was preliminarily explored.Strain B210 prey on different kinds of bacteria,including both gram-stain-positive and gram-stain-negative bacteria,similar like Myxococcus xanthus but different from the members of bdellovibrio,which prey on gram-stain-negative bacteria only.We infer cells connection are necessary for prey phenomenon by observing through scanning electron microscope and cells connection tests.Besides that,we extracted fermentation broth of strain B210 and found it is useless to inhibit bacteria,it proved the necessity of cells connection in another side.