Elimination of macro elements from inoculation and co-cultivation media enhances the efficiency of <Emphasis Type="Italic">Agrobacterium</Emphasis>-mediated transformation in <Emphasis Type="Italic">Petunia</Emphasis>

In order to evaluate the effect of inoculation and co-cultivation media elements on transformation frequency in Petunia hybrida, modified MS media with different elements were tested on Alvan and Large Flower Alvan (LF Alvan), two local cultivars. Leaf explants of both cultivars were inoculated with Agrobacterium tumefaciens strain LBA4404 (pBI121) containing neomycin phosphotransferase (nptII) and an intron-containing β-glucuronidase (gus) genes. When medium lacking KH₂PO₄, NH₄NO₃, KNO₃, and CaCl₂ was used as inoculation and co-cultivation medium, a higher frequency of transformation for Alvan (22%) and LF Alvan (16%) was obtained. Kanamycin resistant plantlets were stained blue by GUS assay. Furthermore, polymerase chain reaction (PCR) analysis revealed the presence of both gus and nptII genes in all putative transformants. Finally, southern blot hybridization confirmed insertion of 1–4 copies of gus gene in transgenic plants.     

Expression of an endochitinase gene from <Emphasis Type="Italic">Trichoderma virens</Emphasis> confers enhanced tolerance to <Emphasis Type="Italic">Alternaria</Emphasis> blight in transgenic <Emphasis Type="Italic">Brassica juncea</Emphasis> (L.) czern and coss lines

An endochitinase gene ‘ech42’ from the biocontrol fungus ‘Trichoderma virens’ was introduced to Brassica juncea (L). Czern and Coss via Agrobaterium tumefaciens mediated genetic transformation method. Integration and expression of the ‘ech42’ gene in transgenic lines were confirmed by PCR, RT-PCR and Southern hybridization. Transgenic lines (T₁) showed expected 3:1 Mendelian segregation ratio when segregation analysis for inheritance of transgene ‘hpt’ was carried out. Fluorimetric analysis of transgenic lines (T₀ and T₁) showed 7 fold higher endochitinase activity than the non-transformed plant. Fluorimetric zymogram showed presence of endochitinase (42 kDa) in crude protein extract of transgenic lines. In detached leaf bioassay with fungi Alternaria brassicae and Alternaria brassicicola, transgenic lines (T₀ and T₁) showed delayed onset of lesions as well as 30–73 % reduction in infected leaf area compared to non-transformed plant.     

Establishment of neomycin phosphotransferase II (<Emphasis Type="Italic">nptII</Emphasis>) selection for transformation of soybean somatic embryogenic cultures

Soybean transformation is limited by the lack of multiple efficient selectable marker systems. Biolistic transformation of somatic proliferative embryogenic cultures, one of the commonly used soybean transformation methods, relies largely on hygromycin phosphotransferase II (hptII) selection. The purpose of the present study was to establish another efficient selectable marker system to facilitate multiple gene transformations of soybean. We tested neomycin phosphotransferase II (nptII) that has been used successfully in cotyledonary node transformation, but with limited success in transformation of embryogenic cultures. Transgenic events were obtained using nptII with improved G418 selection without generating escapes. G418 selection required longer recovery and selection periods, and resulted in a lower efficiency of initial transformants compared to hygromycin selection. Six independent fertile transgenic plants were recovered using nptII and G418, a frequency similar to that obtained with hygromycin selection. Soybean embryogenic cultures co-transformed with the hptII and nptII markers showed resistance to both hygromycin B and G418, while regeneration and plant fertility were not adversely affected. The nptII will be useful as a second selectable marker for multiple gene transformations in basic and applied soybean research.     

Development of leaffolder resistant transgenic rice expressing <Emphasis Type="Italic">cry2AX1</Emphasis> gene driven by green tissue-specific <Emphasis Type="Italic">rbcS</Emphasis> promoter

The insecticidal cry genes of Bacillus thuringiensis (Bt) have been successfully used for development of insect resistant transgenic rice plants. In this study, a novel cry2AX1 gene consisting a sequence of cry2Aa and cry2Ac gene driven by rice rbcS promoter was introduced into a rice cultivar, ASD16. Among 27 putative rice transformants, 20 plants were found to be positive for cry2AX1 gene. The expression of Cry2AX1 protein in transgenic rice plants ranged from 5.95 to 122.40 ng/g of fresh leaf tissue. Stable integration of the transgene was confirmed in putative transformants of rice by Southern blot hybridization analysis. Insect bioassay on T₀ transgenic rice plants against rice leaffolder (Cnaphalocrosis medinalis) recorded larval mortality up to 83.33 %. Stable inheritance and expression of cry2AX1 gene in T₁ progenies was demonstrated using Southern and ELISA. The detached leaf bit bioassay with selected T₁ plants showed 83.33–90.00 % mortality against C. medinalis. The whole plant bioassay for T₁ plants with rice leaffolder showed significant level of resistance even at a lower level of Cry2AX1 expression varying from 131 to 158 ng/g fresh leaf tissue during tillering stage.     

<Emphasis Type="Italic">Nibribacter flagellatus</Emphasis> sp. nov., isolated from rhizosphere of <Emphasis Type="Italic">Hibiscus syriacus</Emphasis> and emended description of the genus <Emphasis Type="Italic">Nibribacter</Emphasis>

A Gram-stain negative, aerobic, motile by flagella, rod-shaped strain (THG-T16^T) was isolated from rhizosphere of Hibiscus syriacus. Growth occurred at 10–40 °C (optimum 28–30 °C), at pH 6.0–8.0 (optimum 7.0) and at 0–1.0% NaCl (optimum 0%). Based on 16S rRNA gene sequence analysis, the near phylogenetic neighbours of strain THG-T16^T were identified as Nibribacter koreensis KACC 16450^T (98.6%), Rufibacter roseus KCTC 42217^T (94.7%), Rufibacter immobilis CCTCC AB 2013351^T (94.5%) and Rufibacter tibetensis CCTCC AB 208084^T (94.4%). The DNA G+C content of strain THG-T16^T was determined to be 46.7 mol%. DNA–DNA hybridization values between strain THG-T16^T and N. koreensis KACC 16450^T, R. roseus KCTC 42217^T, R. immobilis CCTCC AB 2013351^T, R.tibetensis CCTCC AB 208084^T were 33.5?±?0.5% (31.7?±?0.7% reciprocal analysis), 28.1?±?0.2% (25.2?±?0.2%), 17.1?±?0.9% (10.2?±?0.6%) and 8.1?±?0.3% (5.2?±?0.1%). The polar lipids were identified as phosphatidylethanolamine, two unidentified aminophospholipids, an unidentified aminolipid and three unidentified lipids. The quinone was identified as MK-7 and the polyamine as sym-homospermidine. The major fatty acids were identified as C_16:1 ω5c, C_17:1 ω6c, iso-C_15:0, summed feature 3 (C_16:1 ω7c and/or C_16:1 ω6c) and summed feature 4 (iso-C_17:1 I and/or anteiso-C_17:1 B). On the basis of the phylogenetic analysis, chemotaxonomic data, physiological characteristics, and DNA–DNA hybridization data, strain THG-T16^T represents a novel species of the genus Nibribacter, for which the name Nibribacter flagellatus sp. nov. is proposed. The type strain is THG-T16^T(=?KACC 19188^T?=?CCTCC AB 2016246^T).     

Marker-free PLRV resistant potato mediated by Cre-<Emphasis Type="Italic">loxP</Emphasis> excision and RNAi

An inverted repeat construct corresponding to a segment of the potato leaf roll virus coat protein gene was created under control of a constitutive promoter and transferred into a transformation vector with a heat inducible Cre-loxP system to excise the nptII antibiotic resistance marker gene. Fifty-eight transgenic events were evaluated for resistance to PLRV by greenhouse inoculations, which lead to the identification of 7 highly resistant events, of which 4 were extremely resistant. This resistance was also highly effective against accumulation in subsequent tuber generations from inoculated plants, which has not been reported before. Northern blot analysis showed correlation of PLRV specific siRNA accumulation with the level of PLRV resistance. Heat mediated excision of the nptII antibiotic resistance gene in PLRV resistant events was highly efficient in one event with full excision in 71 % of treated explants. On the other hand 8 out of 10 analyzed events showed truncated T-DNA insertions lacking one of the two loxP sites as determined by PCR and confirmed by sequencing flanking regions in 2 events, suggesting cryptic LB sites in the non-coding region between the nptII gene and the flanking loxP site. Accordingly, it is proposed to modify the Cre-loxP vector by reducing the 1 kb size of the region between nptII, loxP, and the LB.     

<Emphasis Type="Italic">Limnobacter humi</Emphasis> sp. nov., a thiosulfate-oxidizing,heterotrophic bacterium isolated from humus soil,and emended description of the genus <Emphasis Type="Italic">Limnobacter</Emphasis> Spring <Emphasis Type="Italic">et al.</Emphasis> 2001

Three Gram-negative, strictly aerobic, chemolithoheterotrophic bacterial strains, designated UCM-30, UCM-33, and UCM-39^T, were isolated in South Korea. Based on their 16S rRNA gene sequences, the three isolated strains were found to be similar to Limnobacter thiooxidans CS-K2^T (97.41–97.68%), Limnobacter litoralis KP1-19^T (95.55–95.76%), and various genera belonging to the class Betaproteobacteria (90.34–93.34%). DNA-DNA hybridization showed 79.3–83.9% similarity between the genomic DNA of UCM-39^T, UCM-30, and UCM-33, while the sequence similarity between UCM-39^T and L. thiooxidans KACC 13837T or L. litoralis LMG 24869T was 23.7% and 18.6%, respectively. The DNA G+C content of UCM 39T was 59.7 mol%, the major ubiquinone was Q-8, and the optimal oxidation rate was observed at 10 mM thiosulfate. The major fatty acids (≥ 10%) were summed features 3 (C_16:1 ω7c and/or C_16:1 ω6c) and 8 (C_18:1 ω7c and/or C_18:1 ω6c), and C_16:0. The major polar lipids (diphosphatidylglycerol, phosphatidylethanolamine, and phosphatidylglycerol) were found in all members of genus Limnobacter. Based on phenotypic, physiological, and phylogenetic analyses, the UCM-39T strain was found to be significantly distinct to represent a novel species affiliated to the genus Limnobacter. We propose to name it Limnobacter humi sp. nov. with the type strain UCM-39^T (=KACC 18574^T =NBRC 111650^T).     

<Emphasis Type="Italic">Flavobacterium parvum</Emphasis> sp. nov., isolated from soil polluted by sewer water

A novel Gram-stain-negative, motile by means of gliding, and short rod-shaped bacterium, designated HS916T, was isolated from soil polluted by sewer water in Cheonan-si, South Korea. Growth occurred at 10–35°C (optimum 30°C), pH 6.0–8.0 (optimum pH 7.0), and 0–1% sodium chloride (NaCl, w/v). Based on similarities of 16S rRNA gene sequences, strain HS916^T was closely related to members of the genus Flavobacterium, exhibiting the highest sequence similarities with Flavobacterium glycines Gm-149^T (96.4%), followed by F. granuli Kw05^T (96.3%), F. fluminis 3R17^T (96.3%), F. aquicola TMd3a3^T (96.2%), and F. nitratireducens N1^T (96.2%). Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain HS916T was placed in a monophyletic cluster with F. nitratireducens N1^T and F. fluminis 3R17^T. The predominant fatty acids (> 5% of the total) of strain HS916^T were iso-C_15:0, anteiso-C_15:0, iso-C_15:0 3-OH, C_17:1ω6с, C_16:0 3-OH, iso-C_17:0 3-OH, and summed feature 3 (C_16:1ω7с and/or C_16:1ω6с). The major polar lipids of the strain comprised phosphatidylethanolamine, unidentified aminolipids, and five unidentified lipids. The predominant respiratory quinone and the major polyamine were menaquinone-6 (MK-6) and symhomospermidine, respectively. The DNA G + C content of strain HS916^T was 34.9 mol%. Based on polyphasic analyses, strain HS916^T represents a novel species belonging to the genus Flavobacterium, for which the name Flavobacterium parvum sp. nov. is proposed. The type strain is HS916T (= KACC 19448^T = JCM 32368^T).     

<Emphasis Type="Italic">Paradevosia shaoguanensis</Emphasis> gen. nov., sp. nov., Isolated from a Coking Wastewater

A Gram staining negative, rod-shaped, aerobic bacterial strain J5-3^T with a single polar flagellum was isolated from coking wastewater collected from Shaoguan, Guangdong, China. It was motile and capable of optimal growth at pH 6–8, 30 °C, and 0–2 % (w/v) NaCl. Its predominant fatty acids were 11-methyl C_18:1 ω7c (29.2 %), C_16:0 (20.6 %), C_19:0 cyclo ω8c (18.2 %), C_18:0 (11.0 %), and C_18:1 ω7c/C_18:1 ω6c (10.9 %) when grown on trypticase soy agar. The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol, two unknown glycolipids (GL1, GL2), and two unknown phospholipid (PL1, PL2). The predominant ubiquinone was Q-10, and the genome DNA G+C content was 61.7 mol %. Phylogenetic analysis based on the 16S rRNA gene sequences indicated that strain J5-3^T belonged to the family Hyphomicrobiaceae in Alphaproteobacteria. It shared the 16S rRNA gene sequence similarities of 93.8–96.1 % with the genus Devosia, 94.5–94.8 % with the genus Pelagibacterium, and <92.0 % with all the other type strains in family Hyphomicrobiaceae. It can be distinguished from the closest phylogenetic neighbors based on several phenotypic and genotypic features, including α-galactosidase activity, tetracycline susceptibility, major fatty acid composition, polar lipid profile, DNA gyrase B subunit (gyrB) gene sequence, and random-amplified polymorphic DNA profile. Therefore, we consider strain J5-3^T to represent a novel species of a novel genus within the family Hyphomicrobiaceae, for which the name Paradevosia shaoguanensis gen. nov., sp. nov. is proposed. The type strain of Paradevosia shaoguanensis is J5-3^T (=CGMCC 1.12430^T =LMG 27409^T).     

<Emphasis Type="Italic">Flavobacterium zaozhuangense</Emphasis> sp. nov., a new member of the family <Emphasis Type="Italic">Flavobacteriaceae</Emphasis>, isolated from metolachlor-contaminated soil

Strain ZZ-8^T, a Gram-negative, aerobic, non-spore-forming, non-motile, yellow-pigmented, rod-shaped bacterium, was isolated from metolachlor-contaminated soil in China. The taxonomic position was investigated using a polyphasic approach. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain ZZ-8^T is a member of the genus Flavobacterium and shows high sequence similarity to Flavobacterium humicola UCM-46^T (97.2%) and Flavobacterium pedocola UCM-R36^T (97.1%), and lower (<?97%) sequence similarity to other known Flavobacterium species. Chemotaxonomic analysis revealed that strain ZZ-8^T possessed MK-6 as the major respiratory quinone; and iso-C_15:0 (28.5%), summed feature 9 (iso-C_17:1 w9c/C_16:0 10-methyl, 22.9%), iso-C_17:0 3-OH (17.0%), iso-C_15:0 3-OH (8.9%), iso-C_15:1 G (8.6%) and summed feature 3 (C_16:1 w7c/C_16:1 w6c, 5.7%) as the predominant fatty acids. The polar lipids of strain ZZ-8^T were determined to be lipids, a glycolipid, aminolipids and phosphatidylethanolamine. Strain ZZ-8^T showed low DNA–DNA relatedness with F. pedocola UCM-R36^T (43.23?±?4.1%) and F. humicola UCM-46^T (29.17?±?3.8%). The DNA G+C content was 43.3 mol%. Based on the phylogenetic and phenotypic characteristics, chemotaxonomic data and DNA–DNA hybridization, strain ZZ-8^T is considered a novel species of the genus Flavobacterium, for which the name Flavobacterium zaozhuangense sp. nov. (type strain ZZ-8^T?=?KCTC 62315 ^T?=?CCTCC AB 2017243^T) is proposed.     

<Emphasis Type="Italic">Corynebacterium defluvii</Emphasis> sp. nov., isolated from Sewage

A Gram-positive, aerobic, non-motile, rod-shapeds, catalase-positive, and oxidase-negative strain, designated Y49^T, was isolated from sewage collected from Jilin Agricultural University, China. It grew at 20–40°C (optimum at 30°C), at pH 6.0–8.0 (optimum at 7.0) and at 0–1.0% sodium chloride (optimum at 0%). The major isoprenoid quinone was menaquinone-8 (MK-8) and the polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylmethylethanolamine, four unidentified lipids, and two unidentified aminolipids. The peptidoglycan was meso-diaminopimelic acid. The cell-wall sugars were galactose, arabinose, and glucose. The fatty acids were C_9:0, C_16:0, C_16:1 ω9c, C_17:1 ω9c, C_18:3 ω6c (6,9,12), C_18:1 ω9c, and C_18:0. The DNA G+C content was 51.4 mol%. Based on the 16S rRNA gene sequence analysis, the nearest phylogenetic neighbors of strain Y49^T were Corynebacterium efficiens DSM 44549^T (97.5%), Corynebacterium callunae DSM 20147^T (97.2%), Corynebacterium deserti GIMN 1.010^T (96.8%), Corynebacterium glutamicum ATCC 13032^T (96.4%), and other species belonging to this genus (92.3–95.4%). The DNA-DNA relatedness value between strain Y49^T and C. efficiens DSM 44549^T, C. callunae DSM 20147^T, C. deserti GIMN1.010^T, and C. glutamicum ATCC 13032^T was 25.5±2.0%, 21.1±1.0%, 16.5±0.5%, and 13.5±0.9%, respectively. Based on the phylogenetic analysis, chemotaxonomic data, physiological characteristics and DNA-DNA hybridization data, strain Y49^T represents a novel species of the genus Corynebacterium, for which the name Corynebacterium defluvii sp nov. is proposed. The type strain is Y49^T (= KCTC 39731^T =CGMCC 1.15506^T).     

<Emphasis Type="Italic">Oceanobacillus gochujangensis</Emphasis> sp. nov., isolated from <Emphasis Type="Italic">gochujang</Emphasis> a traditional Korean fermented food

A Gram-stain-positive, polar flagella-containing, rod-shaped, obligate aerobic, endospore-forming bacterium, strain TK1655^T, was isolated from the traditional Korean food gochujang. The 16S rRNA sequence of strain TK1655^T was a member of the genus Oceanobacillus similar to that of the type strain of Oceanobacillus oncorhynchi subsp. incaldanensis DSM 16557^T (97.2%), O. oncorhynchi subsp. oncorhynchi JCM 12661^T (97.1%), O. locisalsi KCTC 13253^T (97.0%), and O. sojae JCM 15792^T (96.9%). Strain TK1655^T was oxidase and catalase positive. Colonies were circular, smooth, low convex, cream in colour, and measured about 0.5–1.0 mm in diameter. The range for growth was 20–40°C (optimal, 30°C), pH 6.0–10.0 (optimal, 7.0), and 2–16% (w/v) NaCl (optimal, 2%). Additionally, the cells contained meso-DAP, and the predominant isoprenoid quinone was MK-7. The complex polar lipids were consisted of diphosphatidylglycerol (DPG), phosphatidylglycerol (PG), phosphatidylcholine (PC). The major cellular fatty acid components were iso-C_15:0, anteiso-C_15:0, iso-C_16:0, and anteiso-C_17:0, and the DNA G+C content was 40.5%. DNA-DNA relatedness of our novel strain and reference strain O. locisalsi KCTC 13253^T, O. oncorhynchi subsp. incaldanensis DSM 16557^T, O. oncorhynchi subsp. oncorhynchi JCM 12661^T was 45.7, 43.8, and 41.9%. From the results of phenotypic, chemotaxonomic, and phylogenetic analyses of strain TK1655^T, we propose the novel species Oceanobacillus gochujangensis sp. nov. The type strain is TK1655^T (=KCCM 101304^T =KCTC 33014^T =CIP 110582^T =NBRC 109637^T).     

<Emphasis Type="Italic">Scopulibacillus cellulosilyticus</Emphasis> sp. nov., a cellulose-degrading bacterium isolated from tea

A Gram-stain positive, aerobic, non-motile, endospore-forming and rod-shaped strain (THG-NT9^T) was isolated from a green tea sample. Growth occurred at 20–45 °C (optimum 28–35 °C), at pH 6.0–8.0 (optimum 7.0) and at 0–2.0% NaCl (optimum 0%). Based on 16S rRNA gene sequence analysis, the near phylogenetic neighbours of strain THG-NT9^T were identified as Scopulibacillus daqui DSM 28236^T (98.6%), Scopulibacillus darangshiensis DSM 19377^T (97.4%), Pullulanibacillus pueri CGMCC 1.12777^T (96.7%) and Pullulanibacillus camelliae CGMCC 1.15371^T (96.3%). The DNA G?+?C content of strain THG-NT9^T was determined to be 47.5 mol %. DNA–DNA hybridization values between strain THG-NT9^T and S. daqui DSM 28236^T, S. darangshiensis DSM 19377^T, P. pueri CGMCC 1.12777^T, P. camelliae CGMCC 1.15371^T and Pullulanibacillus naganoensis DSM 10191^T were 41.3?±?0.1 (39.4?±?0.4% reciprocal analysis), 39.1?±?0.1 (37.3?±?0.1%), 21.4?±?0.7 (20.1?±?0.3%), 20.7?±?0.1 (20.1?±?0.4%) and 12.1?±?0.2% (8.3?±?0.2%). The polar lipids were identified as diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and three unidentified lipids. The quinone was identified as MK-7. The major fatty acids were C_18:3 ω7c, iso-C_15:0, iso-C_16:0, iso-C_17:0 and anteiso-C_17:0. The cell wall type was determined to be A1γ peptidoglycan with meso-diaminopimelic acid as the diagnostic diamino acid plus alanine and glutamic acid and glucose as the cell wall sugar. On the basis of the phylogenetic analysis, chemotaxonomic data, physiological characteristics, and DNA–DNA hybridization data, strain THG-NT9^T represents a novel species of the genus Scopulibacillus, for which the name Scopulibacillus cellulosilyticus sp. nov. is proposed. The type strain is THG-NT9^T (=?KCTC 33918^T?=?CGMCC 1.16305^T).     

<Emphasis Type="Italic">Hymenobacter sedentarius</Emphasis> sp. nov., isolated from a soil

A novel Gram-negative and red-pinkish bacterium designated DG5B^T was isolated from a dry soil. Cells were rods that were catalase- and oxidase-positive, and non-motile. The strain was found to grow at temperatures from 10 to 30°C (optimum 25°C) and pH 6.0–8.0, (optimum pH 7) on R2A broth. 16S rRNA gene sequence (1,452 bp) analysis of this strain identified it as a member of the genus Hymenobacter that belongs to the class Cytophagia. The highest gene sequence similarities were with Hymenobacter arizonensis OR362-8^T (98.3%), Hymenobacter humi DG31A^T (97.6%), and Hymenobacter glaciei VUG-A130^T (96.6%). Strain DG5B^T exhibited <70% DNA-DNA relatedness with H. arizonensis (34.7 ± 7.0%; reciprocally, 29.7 ± 1.2%) and H. humi (39.4 ± 4.3%; reciprocally, 39.5 ± 3.3%) as a different genomic species, and its genomic DNA G+C content was 59.8%. Strain DG5B^T had the following chemotaxonomic characteristics: the major fatty acids are iso-C_15:0, anteiso-C_15:0, C_16:1ω5c, and summed feature 3 (C_16:1ω7c / C_16:1ω6c); polar lipid profile contained phosphatidylethanolamine (PE), unknown aminophospholipid (APL), unknown glycolipids (GL), unknown phospholipids (PL), and unknown polar lipids (L); the major quinone is MK-7. The absorbance peak of pigment is at 481.0 nm. Strain DG5B^T showed low-level resistance to gamma-ray irradiation. Phenotypic, chemotaxonomic, and genotypic properties indicated that isolate DG5B^T represents a novel species within the genus Hymenobacter for which the name Hymenobacter sedentarius sp. nov. is proposed. The type strain is DG5B^T (=KCTC 32524^T =JCM 19636^T).     

<Emphasis Type="Italic">Halomonas heilongjiangensis</Emphasis> sp. nov., a novel moderately halophilic bacterium isolated from saline and alkaline soil

A moderately halophilic bacterium, designated strain 9-2^T, was isolated from saline and alkaline soil collected in Lindian county, Heilongjiang province, China. The strain was observed to be strictly aerobic, Gram-negative, rod-shaped, oxidase-positive, catalase-positive and motile. It was found to require NaCl for growth and to grow at NaCl concentrations of 0.5–14 % (w/v) (optimum, 7–10 %, w/v), at temperatures of 10–45 °C (optimum 25–30 °C) and at pH 5.0–10.0 (optimum pH 8.0). Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain 9-2^T is a member of the genus Halomonas and is closely related to Halomonas desiderata DSM 9502^T (96.68 %), Halomonas campaniensis DSM 1293^T (96.46 %), Halomonas ventosae DSM 15911^T (96.27 %) and Halomonas kenyensis DSM 17331^T (96.27 %). The DNA–DNA hybridization value was 38.9 ± 0.66 % between the novel isolate 9-2^T and H. desiderata DSM 9502^T. The predominant ubiquinones were identified as Q9 (75.1 %) and Q8 (24.9 %). The major fatty acids were identified as C_16:0 (22.0 %), Summed feature 8 (C_18:1 ω6c/C_18:1 ω7c, 19.6 %), Summed feature 3 (C_16:1 ω6c/C_16:1 ω7c, 12.6 %), C_12:0 3-OH (12.0 %) and C_10:0 (11.7 %). The DNA G+C content was determined to be 69.7 mol%. On the basis of the evidence presented in this study, strain 9-2^T is considered to represent a novel species of the genus Halomonas, for which the name Halomonas heilongjiangensis sp. nov. is proposed. The type strain is 9-2^T (=DSM 26881^T = CGMCC 1.12467^T).     

<Emphasis Type="Italic">Gallaecimonas mangrovi</Emphasis> sp. nov., a novel bacterium isolated from mangrove sediment

A Gram-stain negative, rod-shaped, non-motile, strictly aerobic bacterium HK-28^T was isolated from a mangrove sediment sample in Haikou city, Hainan Province, China. Strain HK-28^T was able to grow at 10–45 °C (optimum 25–30 °C), pH 5.0–8.5 (optimum 6.0–7.0) and 0.5–12.0% (w/v) NaCl (optimum 1.0–3.0%, w/v). The major cellular fatty acids were C_16:0, Summed Feature 8 (C_18:1 ω7c and/or C_18:1 ω6c), Summed Feature 3 (C_16:1 ω7c and/or C_16:1 ω6c), C_17:0, C_12:0 3-OH and C_17:1ω8c. Ubiquinone-8 (Q-8) was the predominant respiratory quinone. The polar lipids consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, two unidentified aminophospholipids, four unidentified phospholipids, two unidentified glycolipid, an unidentified glycophospholipid, an unidentified aminolipid and an unidentified lipid. The DNA G+C content was 50.2 mol%. Accoroding to 16S rRNA gene sequence similarities, strain HK-28^T shared 97.1 and 96.7% sequence similarities to the validly named species Gallaecimonas xiamenensis MCCC 1A01354^T and Gallaecimonas pentaromativorans MCCC 1A06435^T, respectively, and shared lower sequence similarities (<?92.0%) to all other genera. Phylogenetic analysis showed strain HK-28^T was clustered with G. pentaromativorans MCCC 1A06435^T and G. xiamenensis MCCC 1A01354^T. Strain HK-28^T showed low DNA–DNA relatedness with G. xiamenensis MCCC 1A01354^T (28.3?±?1.5%) and G. pentaromativorans MCCC 1A06435^T (25.2?±?2.4%). On the basis of phenotypic, chemotaxonomic and genotypic characteristics, strain HK-28^T is considered to represent a novel species in the genus Gallaecimonas, for which the name Gallaecimonas mangrovi sp. nov. is proposed. The type strain is HK-28^T (=?KCTC 62177^T?=?MCCC 1K03441).     

<Emphasis Type="Italic">Paenibacillus seodonensis</Emphasis> sp. nov., isolated from a plant of the genus <Emphasis Type="Italic">Campanula</Emphasis>

Strain DCT-19^T, representing a Gram-stain-positive, rodshaped, aerobic bacterium, was isolated from a native plant belonging to the genus Campanula on Dokdo, the Republic of Korea. Comparative analysis of the 16S rRNA gene sequence showed that this strain was closely related to Paenibacillus amylolyticus NRRL NRS-290^T (98.6%, 16S rRNA gene sequence similarity), Paenibacillus tundrae A10b^T (98.1%), and Paenibacillus xylanexedens NRRL B-51090^T (97.6%). DNADNA hybridization indicated that this strain had relatively low levels of DNA-DNA relatedness with P. amylolyticus NRRL NRS-290^T (30.0%), P. xylanexedens NRRL B-51090^T (29.0%), and P. tundrae A10b^T (24.5%). Additionally, the genomic DNA G + C content of DCT-19^T was 44.8%. The isolated strain grew at pH 6.0–8.0 (optimum, pH 7.0), 0–4% (w/v) NaCl (optimum, 0%), and a temperature of 15–45°C (optimum 25–30°C). The sole respiratory quinone in the strain was menaquinone-7, and the predominant fatty acids were C_15:0 anteiso, C_16:0 iso, and C_16:0. In addition, the major polar lipids were diphosphatidylglycerol and phosphatidylethanolamine. Based on its phenotypic properties, genotypic distinctiveness, and chemotaxonomic features, strain DCT-19^T is proposed as a novel species in the genus Paenibacillus, for which the name Paenibacillus seodonensis sp. nov. is proposed (=KCTC 43009^T =LMG 30888^T). The type strain of Paenibacillus seodonensis is DCT-19^T.     

<Emphasis Type="Italic">Dankookia rubra</Emphasis> gen. nov., sp. nov., an alphaproteobacterium isolated from sediment of a shallow stream

WS-10^T–a Gram-negative, non-motile, and aerobic bacterial strain–was isolated from the sediment of a shallow stream in Korea. The optimum ranges of temperature and pH for growth were 20–40°C (optimum 28°C) and pH 6.0–8.0 (optimum pH 7.0), respectively. The DNA G+C content of strain WS-10^T was 72.7 mol%. The major fatty acids (>5%) were summed feature 8 (C_18:1ω7c), summed feature 3 (C_16:1ω7c and/or C_16:1ω6c), C_16:0, and C_18:1 2-OH. The major polar lipids consisted of phosphatidylcholine, phosphatidylglycerol, phosphatidylethanolamine, and unidentified aminolipids. Q-10 was the predominant respiratory quinone. The highest similarities in the 16S rRNA gene sequence were shown with Paracraurococcus ruber (95.3%), Belnapia soli (95.3%), B. moabensis (95.1%), and B. rosea (95.0%). A phylogenetic analysis based on 16S rRNA gene sequence comparisons showed that strain WS-10T formed a distinct line within a clade containing the genera Paracraurococcus, Craurococcus, and Belnapia in the family Acetobacteraceae. On the basis of polyphasic evidence, strain WS-10^T represents a novel species of a new genus in the family Acetobacteraceae, for which the name Dankookia rubra gen. nov., sp. nov. is proposed. The type strain of the type species is WS-10^T (= KACC 18533^T = JCM 30602^T).     

<Emphasis Type="Italic">Siphonobacter intestinalis</Emphasis> sp. nov., a bacterium isolated from the feces of <Emphasis Type="Italic">Pseudorhynchus japonicus</Emphasis>

Strain 63MJ-2^T was isolated from the feces of broad-winged katydid (Pseudorhynchus japonicus) collected in Korea. The 16S rRNA gene sequence of this strain showed the highest sequence similarity with that of Siphonobacter aquaeclarae P2^T (96.1%) and had low similarities (below 86.3%) with those of other members of family ‘Flexibacteraceae’. The strain 63MJ-2^T is a strictly aerobic, Gram-stain-negative, non-motile, rod-shaped bacterium. The strain grew at 4–35°C (optimum, 25–30°C), pH of 5.0–9.0 (optimum, 6.0–7.0), and 0–2.0% (optimum, 1.0–2.0) (w/v) NaCl. The DNA G+C content of strain 63MJ-2^T was 43.5 mol%. The major fatty acids were C_16:1ω5c (42.5%), iso-C_17:0 3-OH (18.7%), and summed feature 3 (iso-C_15:0 2-OH and/or C_16:1ω7c, 18.0%). The major menaquinone was MK-7 and polar lipids were phosphatidylethanolamine, six unknown aminolipids, and five unknown lipids. Based on the evidence from our polyphasic taxonomic study, we conclude that strain 63MJ-2^T should be classified as a novel species of the genus Siphonobacter, and propose the name Siphonobacter intestinalis sp. nov. The type strain is 63MJ-2^T (=KACC 18663^T =NBRC 111883^T).