Comparative genomic analysis of bacteriophage EP23 infecting Shigella sonnei and Escherichia coli.

Bacteriophage EP23 that infects Escherichia coli and Shigella sonnei was isolated and characterized. The bacteriophage morphology was similar to members of the family Siphoviridae. The 44,077 bp genome was fully sequenced using 454 pyrosequencing. Comparative genomic and phylogenetic analyses showed that EP23 was most closely related to phage SO-1, which infects Sodalis glossinidius and phage SSL-2009a, which infects engineered E. coli. Genomic comparison indicated that EP23 and SO-1 were very similar with each other in terms of gene order and amino acid similarity, even though their hosts were separated in the level of genus. EP23 and SSL-2009a displayed high amino acid similarity between their genes, but there was evidence of several recombination events in SSL-2009a. The results of the comparative genomic analyses further the understanding of the evolution and relationship between EP23 and its bacteriophage relatives.

Phenotypic characterization and genomic analysis of the Shigella sonnei bacteriophage SP18.

A novel bacteriophage that infects Shigella sonnei was isolated from the Gap River in Korea, and its phenotypic and genomic characteristics were investigated. The virus, called SP18, showed morphology characteristic of the family Myoviridae, and phylogenetic analysis of major capsid gene (gp23) sequences classified it as a T4-like phage. Based on host spectrum analysis, it is lytic to S. sonnei, but not to Shigella flexneri, Shigella boydii or members of the genera Escherichia and Salmonella. Pyrosequencing of the SP18 bacteriophage genome revealed a 170-kb length sequence. In total, 286 ORFs and 3 tRNA genes were identified, and 259 ORFs showed similarity (BLASTP e-value<0.001) to genes of other bacteriophages. The results from comparative genomic analysis indicated that the enterophage JS98, isolated from human stool, is the closest relative of SP18. Based on phylogenetic analysis of gp23 protein-coding sequences, dot plot comparison and BLASTP analysis of genomes, SP18 and JS98 appear to be closely related to T4-even phages. However, several insertions, deletions, and duplications indicate differences between SP18 and JS98. Comparison of duplicated gp24 genes and the soc gene showed that duplication events are responsible for the differentiation and evolution of T4-like bacteriophages.

Agromyces atrinae sp. nov., isolated from fermented seafood.

A Gram-staining-positive, aerobic, non-motile and rod-shaped bacterium, designated P27(T), was isolated from a traditional fermented seafood. The isolate grew optimally with 0-2.0 % (w/v) NaCl and at pH 6-7 and 30 degrees C. The predominant menaquinones were MK-12 and MK-11. The major cellular fatty acids were anteiso-C(17 : 0), anteiso-C(15 : 0) and iso-C(16 : 0). The major cell-wall sugars were galactose, mannose and rhamnose. The peptidoglycan amino acids of strain P27(T) were 2,4-diaminobutyric acid, alanine, glutamic acid and glycine. The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol and an unidentified glycolipid. The genomic DNA G+C content of strain P27(T) was 69.0 mol%. Based on its 16S rRNA gene sequence, strain P27(T) showed highest pairwise similarity with Agromyces cerinus subsp. cerinus JCM 9083(T) (97.0 % similarity). Based on phenotypic, genotypic and phylogenetic studies, strain P27(T) represents a novel species in the genus Agromyces, for which the name Agromyces atrinae sp. nov. is proposed. The type strain is P27(T) (=KCTC 19593(T) =JCM 15913(T)).

Investigation of archaeal and bacterial diversity in fermented seafood using barcoded pyrosequencing.

Little is known about the archaeal diversity of fermented seafood; most of the earlier studies of fermented food have focused on lactic acid bacteria (LAB) in the fermentation process. In this study, the archaeal and bacterial diversity in seven kinds of fermented seafood were culture-independently examined using barcoded pyrosequencing and PCR-denaturing gradient gel electrophoresis (DGGE) methods. The multiplex barcoded pyrosequencing was performed in a single run, with multiple samples tagged uniquely by multiplex identifiers, using different primers for Archaea or Bacteria. Because PCR-DGGE analysis is a conventional molecular ecological approach, this analysis was also performed on the same samples and the results were compared with the results of the barcoded pyrosequencing analysis. A total of 13 372 sequences were retrieved from 15 898 pyrosequencing reads and were analyzed to evaluate the diversity of the archaeal and bacterial populations in seafood. The most predominant types of archaea and bacteria identified in the samples included extremely halophilic archaea related to the family Halobacteriaceae; various uncultured mesophilic Crenarchaeota, including Crenarchaeota Group I.1 (CG I.1a and CG I.1b), Marine Benthic Group B (MBG-B), and Miscellaneous Crenarchaeotic Group (MCG); and LAB affiliated with genus Lactobacillus and Weissella. Interestingly, numerous uncultured mesophilic Crenarchaeota groups were as ubiquitous in the fermented seafood as in terrestrial and aquatic niches; the existence of these Crenarchaeota groups has not been reported in any fermented food. These results indicate that the archaeal populations in the fermented seafood analyzed are diverse and include the halophilic and mesophilic groups, and that barcoded pyrosequencing is a promising and cost-effective method for analyzing microbial diversity compared with conventional approaches.

Haloterrigena jeotgali sp. nov., an extremely halophilic archaeon from salt-fermented food.

A novel red-pigmented halophilic archaeon, strain A29T, was isolated from shrimp jeotgal, a traditional salt-fermented food from Korea. This strain grows in the ranges 10-30% (w/v) NaCl, 17-50 degrees C and pH 6.5-8.5, with optimal growth occurring at 15-20% NaCl, 37-45 degrees C and pH 7.0-7.5. The isolate is Gram-negative and non-motile. Phylogenetic analysis, based on 16S rRNA gene sequences, showed that strain A29T is associated with the genus Haloterrigena and closely related to the species Haloterrigena thermotolerans (99.0% similarity). However, DNA-DNA hybridization experiments revealed that the level of hybridization between strain A29T and related strains of Haloterrigena is less than 70%. The polar lipid fraction consists of phosphatidylglyerol (PG), phosphatidylglycerol phosphate methyl ester (PGP-Me) and mannose-2,6-disulfate(1-2)-glucose glycerol diether (S2-DGD). The G+C content of genomic DNA of the type strain is 62.3 mol%. On the basis of this polyphasic taxonomic study, strain A29T should be placed in the genus Haloterrigena as a novel species, for which the name Haloterrigena jeotgali sp. nov. is proposed. The type strain of the new species is A29T (=KCTC 4020T=DSM 18794T=JCM 14585T=CECT 7218T).

Alishewanella jeotgali sp. nov., isolated from traditional fermented food, and emended description of the genus Alishewanella.

A novel Gram-negative and facultative anaerobic strain, designated MS1T, was isolated from gajami sikhae, a traditional fermented food in Korea made from flatfish. Strain MS1T was motile, rod-shaped and oxidase- and catalase-positive, and required 1-2% (w/v) NaCl for growth. Growth occurred at temperatures ranging from 4 to 40 degrees C and the pH range for optimal growth was pH 6.5-9.0. Strain MS1T was capable of reducing trimethylamine oxide, nitrate and thiosulfate. Phylogenetic analysis placed strain MS1T within the genus Alishewanella. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain MS1T was related closely to Alishewanella aestuarii B11T (98.67% similarity) and Alishewanella fetalis CCUG 30811T (98.04% similarity). However, DNA-DNA reassociation experiments between strain MS1T and reference strains showed relatedness values<70% (42.6 and 14.8% with A. aestuarii B11T and A. fetalis CCUG 30811T, respectively). Genotypic, physiological and biochemical analyses allowed the differentiation of strain MS1T from type strains of species belonging to the genus Alishewanella. Therefore, we propose that strain MS1T (=KCTC 22429T=JCM 15561T) is assigned to a novel species, Alishewanella jeotgali sp. nov.

Sphingomonas aestuarii sp. nov., isolated from tidal flat sediment.

A novel bacterium (strain K4(T)) belonging to the genus Sphingomonas was isolated from tidal flat sediment in Korea. Its morphology, physiology, biochemical features and 16S rRNA gene sequence were characterized. Colonies of this strain are yellow in colour and the cells are rod-shaped, exhibiting negative Gram staining. The strain grows at 0-5 % (w/v) NaCl and 20-35 degrees C, with optimal growth occurring at 0 % (w/v) NaCl and 30 degrees C. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain K4(T) is associated with the genus Sphingomonas. Within the phylogenetic tree, this novel strain shares a branching point with Sphingomonas asaccharolytica Y-345(T), with which it shares 97.3 % 16S rRNA gene sequence similarity. The polyamine pattern predominantly contains the Sphingomonas-specific triamine sym-homospermidine. Combined analysis of 16S rRNA gene sequences, DNA-DNA relatedness, physiological and biochemical test results identified genotypic and phenotypic differences between strain K4(T) and other Sphingomonas species. On the basis of these differentiating features, it is concluded that strain K4(T) (=KCTC 22050(T)=DSM 19475(T)) represents a novel species of the genus Sphingomonas, for which the name Sphingomonas aestuarii sp. nov. is proposed.

Henriciella marina gen. nov., sp. nov., a novel member of the family Hyphomonadaceae isolated from the East Sea.

A bacterial strain, designated Iso4(T), was isolated from the East Sea of Korea and was subjected to a poly-phasic taxonomy study including phenotypic and chemotaxonomic characteristics as well as 16S rRNA gene sequence analysis. Cells of the strain were Gram-negative, motile, non-budding, non-stalked, and strictly aerobic. Strain Iso4(T) grew optimally at 20 degrees C in the presence of 1 approximately 2% (w/v) NaCl and at pH 6.9 approximately 7.6. The major respiratory quinone was Q-10 and the major cellular fatty acids were C(18:1) omega 7c (53.5%), C(17:1) omega 5c (11.7%), C(17:1) omega 6c (8.1%), C(16:0) (7.8%), C(17:0) (4.8%), C(15:0) (2.9%), and C(16:1) omega 5c (2.2%). The DNA G+C content of strain Iso4(T) was 56.2 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain Iso4(T) formed a monophyletic clade in the family Hyphomonadaceae, supported by high bootstrap value and was most closely related to the genus Hyphomonas (92 approximately 94%), a member of marine bacteria in the family. The phenotypic, genotypic, and chemotaxonomic evidences also suggest strain Iso4(T) represents a novel genus and species in the family Hyphomonadaceae, for which the name Henriciella gen. nov., sp. nov. is proposed. The type strain is Iso4(T) (=KCTC 12513(T) =DSM 19595(T) =JCM 15116(T)).

Paracoccus aestuarii sp. nov., isolated from tidal flat sediment.

A Gram-negative micro-organism, designated strain B7(T), was isolated from tidal flat sediment and subjected to a polyphasic taxonomic study involving morphological, physiological, biochemical and 16S rRNA gene sequence analyses. A phylogenetic analysis based on the 16S rRNA gene sequence indicated that strain B7(T) belonged to the genus Paracoccus and was closely related phylogenetically to Paracoccus marcusii MH1(T) (97.5 % sequence similarity), Paracoccus marinus KKL-A5(T) (97.5 %), Paracoccus haeundaensis BC74171(T) (97.3 %), Paracoccus carotinifaciens E-396(T) (97.3 %), Paracoccus homiensis DD-R11(T) (97.2 %), Paracoccus seriniphilus MBT-A4(T) (96.9 %) and other type strains of the genus Paracoccus (95.2-96.7 %). The G+C content of the genomic DNA and the major isoprenoid quinone of the type strain were 62.0 mol% and ubiquinone-10, respectively. The major fatty acid components were C(18 : 1)omega7c (68.9 %) and C(18 : 0) (18.1 %); this profile, with C(18 : 1)omega7c as the predominant fatty acid, was characteristic of members of the genus Paracoccus. The 16S rRNA gene sequence analysis, DNA-DNA hybridization studies and physiological and biochemical tests identified genotypic and phenotypic differences between strain B7(T) and recognized Paracoccus species. On the basis of these data, therefore, strain B7(T) represents a novel species of the genus Paracoccus, for which the name Paracoccus aestuarii sp. nov. is proposed. The type strain is B7(T) (=KCTC 22049(T)=DSM 19484(T)=JCM 15119(T)).

Metatranscriptome analysis of lactic acid bacteria during kimchi fermentation with genome-probing microarrays.

We constructed genome probing microarrays (GPM) that are specific to 39 lactic acid bacteria (LAB) in an effort to monitor microbial diversity and biological activity during the fermentation of kimchi, a traditional Korean vegetable product known to contain various health-promoting and immunity-boosting factors. Metagenomes and metatranscriptomes extracted from periodically sampled kimchi soup were labeled, hybridized and comparatively analyzed using GPMs. Each metatranscriptome was prepared by subtracting 16S rRNA and 23S rRNA from the total RNA, and selectively synthesizing mRNA-specific cDNAs from the rRNA-subtracted samples. Metagenomic analysis revealed 23 LAB related to kimchi fermentation [defined as bacteria with more than a 1% average relative composition (ARC)]. Metatranscriptome analysis revealed that, with the exception of two microorganisms, all LAB probed in the microarray contributed to kimchi fermentation. Moreover, the relative compositions of the major LAB remained unchanged (there was less than a 1.5% difference between the maximum and minimum values) in our metagenome analysis, while our metatranscriptome analysis revealed significant differences in the relative compositions of major LAB during fermentation (relative compositions changed by 2.4% to 9.5%). These data indicate that microorganisms that are less abundant in the flora (those with less than a 5% ARC in the metagenomic analysis) also participated in kimchi fermentation with relatively high activities.

Alishewanella aestuarii sp. nov., isolated from tidal flat sediment, and emended description of the genus Alishewanella.

A Gram-negative strain, B11(T), was isolated from tidal flat sediment in Yeosu, Republic of Korea. Strain B11(T) did not require NaCl for growth and grew between 18 and 44 degrees C. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain B11(T) was associated with the genus Alishewanella and was closely related to the type strain of Alishewanella fetalis (98.3 % similarity). Within the phylogenetic tree, the novel isolate shared a branching point with A. fetalis. Analysis of 16S rRNA gene sequences and DNA-DNA relatedness, as well as physiological and biochemical tests, indicated genotypic and phenotypic differences between strain B11(T) and the type strain of A. fetalis. Thus, strain B11(T) is proposed as a representative of a novel species, Alishewanella aestuarii sp. nov.; the type strain is B11(T) (=KCTC 22051(T) =DSM 19476(T)).

Pseudomonas sabulinigri sp. nov., isolated from black beach sand.

A novel Gram-negative, aerobic, motile, short rod-shaped bacterium, designated J64T, was isolated from black sand collected from Soesoggak, Jeju Island, Korea. Cells grew at 4-37 degrees C, at pH 5.5-10.0 and with 0-10 % NaCl. The strain was found to be oxidase- and catalase-positive. Phylogenetic analyses showed that strain J64T belongs to the genus Pseudomonas, forming a monophyletic group with Pseudomonas pachastrellae, Pseudomonas pertucinogena and 'Pseudomonas denitrificans'. The 16S rRNA gene sequence similarity between strain J64T and type strains of all Pseudomonas species with validly published names was below 96.6 %. Low levels of DNA-DNA relatedness were found with respect to type strains of P. pachastrellae and P. pertucinogena, supporting the classification of strain J64T within a novel species of the genus Pseudomonas. Strain J64T contained C(18 : 1)omega7c (37.2 %), C(16 : 0) (20.4 %), summed feature 3 (17.4 %; comprising iso-C(15 : 0) 2-OH and/or C(16 : 1)omega7c) and C(12 : 0) (7.6 %) as major cellular fatty acids. On the basis of the phenotypic and phylogenetic data, strain J64T represents a novel species of the genus Pseudomonas, for which the name Pseudomonas sabulinigri sp. nov. is proposed. The type strain is J64T (=KCTC 22137T =JCM 14963T).

Nocardioides basaltis sp. nov., isolated from black beach sand.

A novel Gram-positive, aerobic, short-rod-shaped bacterium, designated strain J112T, was isolated from black sand collected from Soesoggak, Jeju Island, Korea. The strain was found to be oxidase-negative and catalase-positive. Cells grew at 10-37 degrees C, at pH 5.5-8.0 and with 1-10 % NaCl. Growth occurred on marine agar but not on R2A or trypticase soy agar. A phylogenetic analysis based on 16S rRNA gene sequences showed that the strain belongs to the radiation of the genus Nocardioides. Strain J112T shared the highest 16S rRNA gene sequence similarities with Nocardioides marinisabuli SBS-12T (99.2 %), Nocardioides terrigena DS-17T (97.3 %), Nocardioides kribbensis KCTC 19038T (97.1 %) and type strains of other Nocardioides species with validly published names (<97 %). The DNA-DNA hybridization values between strain J112T and the three most closely related strains were low enough to justify the assignment of this strain to a novel species. On the basis of these phenotypic, phylogenetic and chemotaxonomic data, strain J112T represents a novel species of the genus Nocardioides, for which the name Nocardioides basaltis sp. nov. is proposed. The type strain is J112T (=KCTC 19365T=JCM 14945T).

Nitratireductor basaltis sp. nov., isolated from black beach sand.

A Gram-negative, coccoid- or rod-shaped bacterium was isolated from black sand collected from Soesoggak beach, Jeju Island, Korea. The isolate, designated J3T, grew at 15-45 degrees C, at pH 5.5-10.0 and in 0-8 % NaCl. It was oxidase- and catalase-positive. Strain J3T reduced nitrate to nitrite, but did not reduce nitrite to nitrogen gas. Phylogenetic analysis of the 16S rRNA gene sequence showed that strain J3T was closely related to Nitratireductor aquibiodomus NL21T and belonged to the genus Nitratireductor. Major cellular fatty acids were C(18 : 1)omega7c (82.0 %), C(19 : 0)omega8c cyclo (4.3 %) and C(18 : 0) (4.0 %), a profile that is typical of members of the genus Nitratireductor and distinct from those of other genera in the family Phyllobacteriaceae. Differences in physiological characteristics and fatty acid profiles, as well as low DNA-DNA hybridization values, further established that strain J3T was distinct from N. aquibiodomus NL21T. Thus, strain J3T (=KCTC 22119T =JCM 14935T) should be classified as the type strain of a novel species in the genus Nitratireductor, for which the name Nitratireductor basaltis sp. nov. is proposed.

Application of quantitative real-time PCR for enumeration of total bacterial, archaeal, and yeast populations in kimchi.

Kimchi is a Korean traditional fermented food made of brined vegetables, with a variety of spices. Various microorganisms are associated with the kimchi fermentation process. This study was undertaken in order to apply quantitative real-time PCR targeting the 16S and 26S rRNA genes for the investigation of dynamics of bacterial, archaeal, and yeast communities during fermentation of various types of kimchi. Although the total bacterial and archaeal rRNA gene copy numbers increased during kimchi fermentation, the number of yeasts was not significantly altered. In 1 ng of bulk DNA, the mean number of rRNA gene copies for all strains of bacteria was 5.45 x 10(6) which was 360 and 50 times greater than those for archaea and yeast, respectively. The total gene copy number for each group of microorganisms differed among the different types of kimchi, although the relative ratios among them were similar. The common dominance of bacteria in the whole microbial communities of various types of kimchi suggests that bacteria play a principal role in the kimchi fermentation process.

Marinomonas basaltis sp. nov., a marine bacterium isolated from black sand.

A Gram-negative, aerobic, slightly halophilic, rod-shaped bacterium was isolated from black sand in Soesoggak, Jeju island, Korea. The strain, designated J63(T), was oxidase- and catalase-positive and arginine dihydrolase-negative. The isolate required Na(+) for growth and differed from phenotypically related species by being able to utilize sucrose and d-galactose as a carbon source. Phylogenetic analysis based on the sequence of the 16S rRNA gene revealed that strain J63(T) belongs to the genus Marinomonas. It exhibited 16S rRNA gene sequence similarities of 97.6-98.7 % to the closely related species Marinomonas communis, Marinomonas ostreistagni, Marinomonas aquimarina and Marinomonas vaga. The phylogenetic analysis revealed that strain J63(T) comprised a relatively long subline of descent, shared a branch point with the outlying species Marinomonas communis and occupied a phylogenetically distant position on the main Marinomonas branch. Based on DNA-DNA hybridization, the levels of relatedness between strain J63(T) and M. communis NBRC 102224(T), M. aquimarina CIP 108405(T) and M. vaga JCM 20774(T) were 56.2, 45.1 and 51.3 %, respectively. On the basis of the phenotypic, genetic and phylogenetic data, strain J63(T) should be placed in the genus Marinomonas as representing a novel species, for which the name Marinomonas basaltis sp. nov. is proposed. The type strain is J63(T) (=KCTC 22118(T)=JCM 14948(T)).

Marinobacter goseongensis sp. nov., from seawater.

A Gram-negative marine bacterium, designated strain En6(T), was isolated from seawater of the East Sea of Korea. The organism grew in 1-25 % (w/v) NaCl and at 10-37 degrees C and pH 5.3-9.3, with optimal growth occurring in 4-5 % NaCl and at 25-30 degrees C and pH 7.5. Phylogenetic analysis of the 16S rRNA gene sequence of strain En6(T) placed this bacterium in the clade Marinobacter within the class Gammaproteobacteria. The 16S rRNA gene sequence similarity between strain En6(T) and Marinobacter lipolyticus SM19(T), the most closely related species, was 98.4 %, and the level of DNA-DNA relatedness between the two strains was 22 %. On the basis of the phylogenetic analysis and phenotypic and chemotaxonomic data, strain En6(T) is considered to represent a novel species of the genus Marinobacter. The name Marinobacter goseongensis sp. nov. is proposed, with strain En6(T) (=KCTC 12515(T)=DSM 19471(T)) as the type strain.

Bacterial, archaeal, and eukaryal diversity in the intestines of Korean people.

The bacterial, archaeal, and eukaryal diversity in fecal samples from ten Koreans were analyzed and compared by using the PCR-fingerprinting method, denaturing gradient gel electrophoresis (DGGE). The bacteria all belonged to the Firmicutes and Bacteroidetes phyla, which were known to be the dominant bacterial species in the human intestine. Most of the archaeal sequences belonged to the methane-producing archaea but several halophilic archarea-related sequences were also detected unexpectedly. While a small number of eukaryal sequences were also detected upon DGGE analysis, these sequences were related to fungi and stramenopiles (Blastocystis hominis). With regard to the bacterial and archaeal DGGE analysis, all ten samples had one and two prominent bands, respectively, but many individual-specific bands were also observed. However, only five of the ten samples had small eukaryal DGGE bands and none of these bands was observed in all five samples. Unweighted pair group method and arithmetic averages clustering algorithm (UPGMA) clustering analysis revealed that the archaeal and bacterial communities in the ten samples had relatively higher relatedness (the average Dice coefficient values were 68.9 and 59.2% for archaea and bacteria, respectively) but the eukaryal community showed low relatedness (39.6%).

Luteimonas aestuarii sp. nov., isolated from tidal flat sediment.

A novel bacterium B9(T) was isolated from tidal flat sediment. Its morphology, physiology, biochemical features, and 16S rRNA gene sequence were characterized. Colonies of this strain are yellow and the cells are Gram-negative, rod-shaped, and do not require NaCl for growth. The 16S rRNA gene sequence similarity indicated that strain B9(T) is associated with the genus Lysobacter (< or = 97.2%), Xanthomonas (< or = 96.8%), Pseudomonas (< or = 96.7%), and Luteimonas (< or = 96.0%). However, within the phylogenetic tree, this novel strain shares a branching point with the species Luteimonas composti CC-YY255(T) (96.0%). The DNA-DNA hybridization experiments showed a DNA-DNA homology of 23.0% between strain B9(T) and Luteimonas mephitis B1953/27.1(T). The G+C content of genomic DNA of the type strain is 64.7 mol% (SD, 1.1). The predominant fatty acids are iso-C(11:0), iso-C(15:0), iso-C(16:0), iso-C(17:0), iso-C(17:0) omega9c, and iso-C(11:0) 3-OH. Combined analysis of the 16S rRNA gene sequences, fatty acid profile, and results from physiological and biochemical tests indicated that there is genotypic and phenotypic differentiation of the isolate from other Luteimonas species. For these reasons, strain B9(T) was proposed as a novel species, named Luteimonas aestuarii. The type strain of the new species is B9(T) (= KCTC 22048(T), DSM 19680(T)).

Aliihoeflea aestuarii gen. nov., sp. nov., a novel bacterium isolated from tidal flat sediment.

A novel Gram-negative and rod-shaped bacterium, designated N8(T), was isolated from tidal flat sediment. Phylogenetic analysis based on 16S rRNA gene sequences showed that N8(T) strain is associated with the family Phyllobacteriaceae: two uncultured clones (98.4 and 99.8% 16S rRNA gene sequence similarity) and the genus Mesorhizobium (< or =97.0%). The novel strain formed a separate clade with uncultured clones in the phylogenetic tree based on 16S rRNA gene sequences. Cellular fatty acid profiles predominately comprised C(18:1) omega7c and C(19:0) cyclo omega8c. The major isoprenoid quinone is ubiquinone-10 and genomic DNA G+C content is 53.4 mol%. The polyphasic taxonomic study indicates that the novel strain N8(T) represents a novel species of the new genus in the family Phyllobacteriaceae, named Aliihoeflea aestuarii. The type strain is N8(T) (= KCTC 22052(T)= JCM 15118(T)= DSM 19536(T)).