Structural basis of amine odorant perception by a mammal olfactory receptor.

Odorants are detected as smell in the nasal epithelium of mammals by two G-protein-coupled receptor families, the odorant receptors and the trace amine-associated receptors1,2 (TAARs). TAARs emerged following the divergence of jawed and jawless fish, and comprise a large monophyletic family of receptors that recognize volatile amine odorants to elicit both intraspecific and interspecific innate behaviours such as attraction and aversion3-5. Here we report cryo-electron microscopy structures of mouse TAAR9 (mTAAR9) and mTAAR9-Gs or mTAAR9-Golf trimers in complex with ß-phenylethylamine, N,N-dimethylcyclohexylamine or spermidine. The mTAAR9 structures contain a deep and tight ligand-binding pocket decorated with a conserved D3.32W6.48Y7.43 motif, which is essential for amine odorant recognition. In the mTAAR9 structure, a unique disulfide bond connecting the N terminus to ECL2 is required for agonist-induced receptor activation. We identify key structural motifs of TAAR family members for detecting monoamines and polyamines and the shared sequence of different TAAR members that are responsible for recognition of the same odour chemical. We elucidate the molecular basis of mTAAR9 coupling to Gs and Golf by structural characterization and mutational analysis. Collectively, our results provide a structural basis for odorant detection, receptor activation and Golf coupling of an amine olfactory receptor.

Pillar[n]arene-Based Fluorescence Turn-On Chemosensors for the Detection of Spermine, Spermidine, and Cadaverine in Saline Media and Biofluids.

Polyamines are essential analytes due to their critical role in various biological processes and human health in general. Due to their role as regulators for cell growth and proliferation (putrescine and spermine), as neuroprotectors, gero-, and cardiovascular protectors (spermidine), and as bacterial growth indicators (cadaverine), rapid, simple, and cost-effective methods for polyamine detection in biofluids are in demand. The present study focuses on the development and investigation of self-assembled and fluorescent host⋅dye chemo-sensors based on sulfonated pillar[5]arene for the specific detection of polyamines. Binding studies, as well as stability and functionality assessments of the turn-on chemosensors for selective polyamine detection in saline and biologically relevant media, are shown. Furthermore, the practical applicability of the developed chemo-sensors is demonstrated in biofluids such as human urine and saliva.

Pentafluoropropionic Anhydride Derivatization and GC-MS Analysis of Histamine, Agmatine, Putrescine, and Spermidine: Effects of Solvents and Starting Column Temperature.

Gas chromatography-mass spectrometry (GC-MS) is useful for the quantitative determination of the polyamines spermidine (SPD) and putrescine (PUT) and of the biogenic amine agmatine (AGM) in biological samples after derivatization. This GC-MS method involves a two-step extraction with n-butanol and hydrochloric acid, derivatization with pentafluoropropionic anhydride (PFPA) in ethyl acetate, and extraction of the pentafluoropropionic (PFP) derivatives by toluene of SPD, PUT, and AGM. We wanted to extend this GC-MS method for the biogenic amine histamine (HA), but we faced serious problems that did not allow reliable quantitative analysis of HA. In the present work, we addressed this issue and investigated the derivatization of HA and the effects of toluene and ethyl acetate, two commonly used water-insoluble organic solvents in GC-MS, and oven temperature program. Derivatization of unlabelled HA (d0-HA) and deuterium-labelled HA (d4-HA) with PFPA in ethyl acetate (PFPA-EA, 1:4, v/v; 30 min, 65 °C) resulted in the formation of d0-HA-(PFP)2 and d4-HA-(PFP)2 derivatives. d4-HA and 13C4-SPD were used as internal standards for the amines after standardization. Considerable quantitative effects of toluene and ethyl acetate were observed. The starting GC column temperature was also found to influence considerably the GC-MS analysis of HA. Our study shows the simultaneous quantitative analysis of HA as HA-(PFP)2, AGM as AGM-(PFP)3, PUT as PUT-(PFP)2, and SPD as SPD-(PFP)3 derivatives requires the use of ethyl acetate for their extraction and injection into the GC-MS apparatus and a starting GC column temperature of 40 °C instead of 70 °C. The PFP derivatives of HA, AGM, PUT, and SPD were found to be stable in ethyl acetate for several hours at room temperature. Analytically satisfactory linearity, precision, and accuracy were observed for HA, AGM, PUT, and SPD in biologically relevant ranges (0 to 700 pmol). The limits of detection of AGM, PUT, and SPD were about two times lower in ethyl acetate compared to toluene (range, 1-22 fmol). The limits of detection were 1670 fmol for d0-HA and 557 fmol for d4-HA. Despite the improvements achieved in the study for HA, its analysis by GC-MS as a PFP derivative is challenging and less efficient than that of PUT, AGM, and SPD.

Genetic mapping of sexually dimorphic volatile and non-volatile floral secondary chemistry of a dioecious willow.

Secondary chemistry often differs between sexes in dioecious plant species, a pattern attributed to its possible role in the evolution and/or maintenance of dioecy. We used GC-MS to measure floral volatiles emitted from, and LC-MS to quantitate non-volatile secondary compounds contained in, female and male Salix purpurea willow catkins from an F2 family. Using the abundance of these chemicals, we then performed quantitative trait locus (QTL) mapping to locate them on the genome, identified biosynthetic candidate genes in the QTL intervals, and examined expression patterns of candidate genes using RNA-seq. Male flowers emitted more total terpenoids than females, but females produced more benzenoids. Male tissue contained greater amounts of phenolic glycosides, but females had more chalcones and flavonoids. A flavonoid pigment and a spermidine derivative were found only in males. Male catkins were almost twice the mass of females. Forty-two QTL were mapped for 25 chemical traits and catkin mass across 16 of the 19 S. purpurea chromosomes. Several candidate genes were identified, including a chalcone isomerase associated with seven compounds. A better understanding of the genetic basis of the sexually dimorphic chemistry of a dioecious species may shed light on how chemically mediated ecological interactions may have helped in the evolution and maintenance of dioecy.

Influence of Brewing Process on the Profile of Biogenic Amines in Craft Beers.

The evaluation of the biogenic amines (BAs) profile of different types of craft beers is herein presented. A previously developed and validated analytical method based on ion-pair chromatography coupled with potentiometric detection was used to determine the presence of 10 BAs. Good analytical features were obtained for all amines regarding linearity (R2 values from 0.9873 ± 0.0015 to 0.9973 ± 0.0015), intra- and inter-day precision (RSD lower than 6.9% and 9.7% for beer samples, respectively), and accuracy (recovery between 83.2-108.9%). Detection and quantification limits range from 9.3 to 60.5 and from 31.1 to 202.3 µg L-1, respectively. The validated method was applied to the analysis of four ale beers and one lager craft beer. Ethylamine, spermidine, spermine, and tyramine were detected in all analyzed samples while methylamine and phenylethylamine were not detected. Overall, pale ale beers had a significantly higher total content of BAs than those found in wheat pale and dark samples. A general least square regression model showed a good correlation between the total content of BAs and the brewing process, especially for Plato degree, mashing, and fermentation temperatures. Knowledge about the type of ingredients and manufacturing processes that contribute to higher concentrations of these compounds is crucial to ensuring consumer safety.

Intervening in sibling competition for assimilates by controlled pollination prevents seed abortion under postpollination drought in maize.

During maize production, drought throughout the flowering stage usually induces seed abortion and yield losses. The influence of postpollination drought stress on seed abortion and its underlying mechanisms are not well characterized. By intervening in the competition for assimilates between kernel siblings under different degrees of postpollination drought stresses accompanied by synchronous pollination (SP) and incomplete pollination (ICP) approaches, the mechanisms of postpollination abortion were investigated at physiological and molecular levels. Upon SP treatment, up to 15% of the fertilized apical kernels were aborted in the drought-exacerbated competition for assimilates. The aborted kernels exhibited weak sucrose hydrolysis and starch synthesis but promoted the synthesis of trehalose-6-phosphate and ethylene. In ICP where basal pollination was prevented, apical kernel growth was restored with reinstated sucrose metabolism and starch synthesis and promoted sucrose and hexose levels under drought stress. In addition, the equilibrium between ethylene and polyamine in response to the drought and pollination treatments was associated with the abortion process. We conclude that competition for assimilates drives postpollination kernel abortion, whereas differences in sugar metabolism and the equilibrium between ethylene and polyamines may be relevant to the "live or die" choice of kernel siblings during this competition.

Sphingomonas palmae sp. nov. and Sphingomonas gellani sp. nov., endophytically associated phyllosphere bacteria isolated from economically important crop plants.

In this study, two endophytic bacterial strains designated JS21-1T and S6-262T isolated from leaves of Elaeis guineensis and stem tissues of Jatropha curcas respectively, were subjected for polyphasic taxonomic approach. On R2A medium, colonies of strains JS21-1T and S6-262T are orange and yellow, respectively. Phylogenetic analyses using 16S rRNA gene sequencing and whole-genome sequences placed the strains in distinct clades but within the genus Sphingomonas. The DNA G + C content of JS21-1T and S6-262T were 67.31 and 66.95%, respectively. Furthermore, the average nucleotide identity and digital DNA-DNA hybridization values of strains JS21-1T and S6-262T with phylogenetically related Sphingomonas species were lower than 95% and 70% respectively. The chemotaxonomic studies indicated that the major cellular fatty acids of the strain JS21-1T were summed feature 8 (C18:1 ω7c and/or C18:1 ω6c), C16:0, and C14:0 2OH; strain S6-262T possessed summed feature 3 (C16:1 ω7c and/or iso-C15:0 2-OH) and summed feature 8 (C18:1 ω6c and/or C18:1 ω7c). The major quinone was Q10, and the unique polyamine observed was homospermidine. The polar lipid profile comprised of mixture of sphingoglycolipid, phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol and certain uncharacterised phospholipids and lipids. Based on this polyphasic evidence, strains JS21-1T and S6-262T represent two novel species of the genus Sphingomonas, for which the names Sphingomonas palmae sp. nov. and Sphingomonas gellani sp. nov. are proposed, respectively. The type strain of Sphingomonas palmae sp. nov. is JS21-1T (= DSM 27348T = KACC 17591T) and the type strain of Sphingomonas gellani sp. nov. is S6-262T (= DSM 27346T =  KACC 17594T).

Regulation of the biosynthesis of endogenous nitric oxide and abscisic acid in stored peaches by exogenous nitric oxide and abscisic acid.

BACKGROUND: Nitric oxide (NO) and abscisic acid (ABA) are important regulators of plant response to cold stress, and they interact in response to cold signals. The primary goal of this study was to determine the roles of exogenous NO and ABA on the synthesis of endogenous NO and ABA in cold-stored peach fruit. RESULTS: Exogenous NO and ABA maintained a relatively high content of NO, increased nitrate reductase (NR) activity, and inhibited the activity of NO synthase (NOS)-like and the levels of polyamine biosynthesis in peaches during cold storage. Treatments of potassium 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (c-PTIO), NO, N-nitro-l-Arg-methyl ester (L -NAME), and sodium tungstate did not influence ABA content. Exogenous ABA increased the content of carotenoids and the activities of aldehyde oxidase (AO), 9-cis-epoxycarotenoid dioxygenase (NCED), and zeaxanthin epoxidase (ZEP) of ABA synthesis in peaches during cold storage, and upregulated the gene expression of PpAO1, PpNCED1, PpNCED2, and PpZEP. The production of endogenous NO was differentially inhibited by NO scavengers, ABA inhibitors, and NR inhibitors, but not affected by NOS-like inhibitors during cold storage. CONCLUSION: Exogenous NO and ABA can induce endogenous NO synthesis in cold-stored peaches by the nitrate reductase pathway, and ABA can mediate endogenous ABA synthesis by the autocatalytic reaction. NO does not regulate ABA synthesis. © 2019 Society of Chemical Industry.

Aestuariisphingobium litorale gen. nov., sp. nov., a novel proteobacterium isolated from a water sample of Pearl River estuary.

Strain SYSU M10002T was isolated from a water sample collected from the coastal region of Pearl River estuary, Guangdong Province, southern China. The taxonomic position of the isolate was investigated by polyphasic taxonomic approaches. The isolate was found to be Gram-negative, non-motile, short rods and aerobic. The strain was able to grow at 14-37 °C, pH 6.0-10.0 and in the presence of up to 0.5% (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain SYSU M10002T is a member of the family Sphingomonadaceae, with high sequence similarity to Sphingorhabdus buctiana T5T (95.1%). Overall genomic related indices between the genome of strain SYSU M10002T and those of related strains were low to moderate (AAI values < 64.3%; POCP values < 58%), indicating that strain SYSU M10002T represents a novel lineage within the family Sphinogomonadaceae. Strain SYSU M10002T contained homospermidine as its polyamine. The major polar lipids were diphosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, sphingoglycolipid, two unidentified phospholipids and an unidentified aminolipid. Ubiquinone Q-9 (44.9%) and Q-10 (43.2%) were the dominant respiratory quinones, along with a minor amount of Q-8 (11.9%). The predominant cellular fatty acids (> 10%) identified were summed feature 3 (C16:1ω7c and/or C16:1ω6c), summed feature 8 (C18:1ω7c) and C14:0 2-OH. The genomic DNA G+C content was 64.0%. Based on the analyses of the phenotypic, genotypic and phylogenetic characteristics, strain SYSU M10002T is determined to represent a novel species of a novel genus, for which the name Aestuariisphingobium litorale gen. nov., sp. nov. is proposed. The type strain of the species is SYSU M10002T (= KCTC 52944T = NBRC 112961T).

Reduction of Spermidine Content Resulting from Inactivation of Two Arginine Decarboxylases Increases Biofilm Formation in Synechocystis sp. Strain PCC 6803.

The phototropic bacterium Synechocystis sp. strain PCC 6803 is able to adapt its morphology in order to survive in a wide range of harsh environments. Under conditions of high salinity, planktonic cells formed cell aggregates in culture. Further observations using crystal violet staining, confocal laser scanning microscopy, and field emission-scanning electron microscopy confirmed that these aggregates were Synechocystis biofilms. Polyamines have been implicated in playing a role in biofilm formation, and during salt stress the content of spermidine, the major polyamine in Synechocystis, was reduced. Two putative arginine decarboxylases, Adc1 and Adc2, in Synechocystis were heterologously expressed in Escherichia coli and purified. Adc2 had high arginine decarboxylase activity, whereas Adc1 was much less active. Disruption of the adc genes in Synechocystis resulted in decreased spermidine content and formation of biofilms even under nonstress conditions. Based on the characterization of the adc mutants, Adc2 was the major arginine decarboxylase whose activity led to inhibition of biofilm formation, and Adc1 contributed only minimally to the process of polyamine synthesis. Taken together, in Synechocystis the shift from planktonic lifestyle to biofilm formation was correlated with a decrease in intracellular polyamine content, which is the inverse relationship of what was previously reported in heterotroph bacteria.IMPORTANCE There are many reports concerning biofilm formation in heterotrophic bacteria. In contrast, studies on biofilm formation in cyanobacteria are scarce. Here, we report on the induction of biofilm formation by salt stress in the model phototrophic bacterium Synechocystis sp. strain PCC 6803. Two arginine decarboxylases (Adc1 and Adc2) possess function in the polyamine synthesis pathway. Inactivation of the adc1 and adc2 genes leads to biofilm formation even in the absence of salt. The shift from planktonic culture to biofilm formation is regulated by a decrease in spermidine content in Synechocystis This negative correlation between biofilm formation and polyamine content, which is the opposite of the relationship reported in other bacteria, is important not only in autotrophic but also in heterotrophic bacteria.

Expression and distribution patterns of spermine, spermidine, and putrescine in rat hair follicle.

No expression and distribution patterns of polyamines (PAs), spermine, spermidine, and their precursor putrescine in mammalian hair follicle are available, although polyamines are known to correlate well with hair growth and epidermal tumor genesis. Immunohistochemistry (IHC) using our original two monoclonal antibodies (mAbs) ASPM-29 specific for spermine or spermidine, and APUT-32 specific for putrescine allowed us to detect immunoreactivity for polyamines in hair follicles from normal adult rats. A wide range of immunoreactivity for the total spermine and spermidine was observed in the compartments of hair follicle: The highest degree of immunoreactivity for polyamines was observed in the matrix, in the Huxley's layer, in the deeper Henle's layer, and in the cuticle of the inner root sheath/the hair cuticle, while moderate immunoreactivity existed in the lower-to-mid cortex and the companion layer, followed by lower immunoreactivity in the outer root sheath, including the bulge region and in the deeper medulla, in which the immunoreactivity was also evident in their nuclei. In addition, somewhat surprisingly, with IHC by APUT-32 mAb, we detected significant levels of putrescine in the compartments, in which the immunostaining pattern was the closely similar to that of the total spermine and spermidine. Thus, among these compartments, the cell types of the matrix, the Huxley's layer, the deeper Henle's layer, and the cuticle of the inner root sheath/the hair cuticle seem to have the biologically higher potential in compartments of anagen hair follicle, maybe suggesting that they are involved more critically in the biological event of hair growth. In addition, we noted sharp differences of immunostaining by IHCs between ASPM-29 mAb and APUT-32 mAb in the epidermis cells and fibroblast. ASPM-29 mAb resulted in strong staining in both the cell types, but APUT-32 mAb showed only very light staining in both types. Consequently, the use of the two IHCs could be extremely useful in further studies on hair cycle and epidermal tumor genesis experimentally or clinically.

Quantitative determination of linking number differences between circular polynucleosomes and histone H1-bound circular polynucleosomes.

With the aim of discovering contribution of histone H1 to linking number changes of DNA, determination of linking number differences between histone H1-free circular polynucleosomes and histone H1-bound circular polynucleosomes was carried out during our investigations. Our results showed that on average, binding of ∼11.5 histone H1 molecules causes one linking number change in circular polynucleosomes in the presence of 1.5 mM spermidine. When concentrations of spermidine decreases or increases, these linking number differences decrease significantly. It is therefore evident that linking number changes caused by histone H1 are spermidine concentration-dependent.

Rapid Evaluation of Spermidine from 12 Bean Cultivars by Direct Real-Time Mass Spectrometry Analysis.

The routine spermidine (SPD) detection method is time-consuming and laborious due to the lengthy chromatographic separation and/or tedious sample derivatization pretreatment. In this study, direct analysis in real-time ionization mode coupled with mass spectrometry (DART-MS) was developed to rapidly determine the SPD content of 12 bean cultivars. The results were compared in detail with those of the classical UHPLC-ESI-QTOF method. After conducting a series of optimizations, a simple sample extraction procedure employing 80% aqueous methanol, was followed by determination of sample extracts directly without any chromatographic separation or prior derivatization. The validated method showed excellent performance with low limits of detection (LOD of 0.025 mg·kg-1) and good recovery rates (102.79⁻148.44%). The investigation highlighted that the DART-MS method (~1.3 min per three samples) could be used as a high-throughput alternative to the classic UHPLC-ESI-QTOF method (~15 min per three samples).

Progressive response of large intestinal bacterial community and fermentation to the stepwise decrease of dietary crude protein level in growing pigs.

The study aimed to determine the effects of reduction of dietary crude protein (CP) level with balanced essential amino acids (EAA) on intestinal bacteria and their metabolites of growing pigs. Forty pigs (initial BW 13.50 ± 0.50 kg, 45 ± 2 days of age) were randomly assigned to four dietary treatments containing CP levels at 20.00% (normal crude protein, NP); 17.16% (medium crude protein, MP); 15.30% (low crude protein, LP); and 13.90% (extremely low crude protein, ELP), respectively. Crystalline AAs were added to meet the EAA requirement of pigs. After 4-week feeding, eight pigs per treatment (n = 8) were randomly selected and slaughtered for sampling of ileal, cecal, and colonic digesta and mucosa. Pigs with moderately reduced CP level had increased bacterial diversity, with the Shannon diversity indices for the colon digesta in the LP group and mucosa in the MP and LP groups significantly (P < 0.05) higher than those in the NP and ELP groups. As the CP level reduces, the Bifidobacterium population were linearly decreased (P < 0.05) both in ileum, cecum, and colon, and the ELP group had the lowest Bifidobacterium population in the cecum and colon, with its value significantly lower than NP and MP groups (P < 0.05). However, the ELP group had the highest population of Escherichia coli in the colon, with its value significantly higher than the LP group (P < 0.05). For bacterial metabolites, as CP level decreased, total short-chain fatty acid (T-SCFA), acetate, and butyrate were linearly increased (linear, P < 0.05) in the ileum, while all SCFAs except formate in the cecum and T-SCFA and acetate in the colon, were linearly decreased (P < 0.05). Reducing CP level led to a linear decrease of microbial crude protein (MCP) in the ileum (P < 0.05) and ammonia in all intestine segments (P < 0.05). The spermidine in cecum and total amines, cadaverine, methylamine, and spermidine in colon were shown a quadratic change (P < 0.05) as dietary CP decreases, with the highest concentration in LP group. These findings suggest that moderate reduction of dietary CP level may benefit large intestinal bacterial community and its fermentation, which was negatively affected by extremely low CP diet.

[Simultaneous determination of four coumaroylspermidine constituents in Carthamus tinctorius by HPLC-DAD].

The HPLC-DAD method was established to simultaneously determine the contents of four coumaroylspermidine[ N1, N5, N10-(Z)-tri-p-coumaroylspermidine (1), N1, N5-(Z)-N10-(E)-tri-p-coumaroylspermidine (2), N1(E)-N5-(Z)-N10-(E)-tri-p-coumaroylspermidine (3), and N1, N5, N10-(E)-tri-p-coumaroyl-spermidine (4) ] in Carthamus tinctorius. The method was performed on an Eclipse XDB-C18 column (4.6 mm×250 mm, 5 µm) eluted with 47% methanol in an isocratic program. The flow rate was 1 mL•min⁻¹; the injection volume was 10 µL, and the column temperature was 30 ℃. The detective wavelength was 270, 280, 290, and 300 nm, respectively. Four coumaroylspermidine constituents showed a good linearity in the range of 0.002 1-0.041 6 (r=0.999 5), 0.002 6-0.051 2 (r=0.999 7), 0.002 7-0.054 0 (r=0.999 8) g•L⁻¹, and 0.005 0-0.100 4 (r=0.999 8) g•L⁻¹, respectively. The average recoveries of these four coumaroylspermidine constituents were in the range of 98.61%-100.9% (RSD 2.3%-3.0%). In conclusion, the method is simple, rapid, and sensitive, which could be used as a quantitative determination method for the four coumaroylspermidine components in C.tinctorius.

Amine oxidase-based biosensors for spermine and spermidine determination.

The present work describes the development and optimization of electrochemical biosensors for specific determination of the biogenic polyamine spermine (Spm) and spermidine (Spmd) whose assessment represents a novel important analytical tool in food analysis and human diagnostics. These biosensors have been prepared using novel engineered enzymes: polyamine oxidase (PAO) endowed with selectivity towards Spm and Spmd and spermine oxidase (SMO) characterized by strict specificity towards Spm. The current design entails biosensors in which the enzymes were entrapped in poly(vinyl alcohol) bearing styrylpyridinium groups (PVA-SbQ), a photocrosslinkable gel, onto an electrode surface. Screen-printed electrodes (SPEs) were used as electrochemical transducers for enzymatically produced hydrogen peroxide, operating at different potential vs Ag/AgCl according to the material of the working electrode (WE): +700 mV for graphite (GP) or -100 mV for Prussian blue (PB)-modified SPE, respectively. Biosensor performances were evaluated by means of flow injection amperometric (FIA) measurements. The modified electrodes showed good sensitivity, long-term stability and reproducibility. Under optimal conditions, the PAO biosensor showed a linear range 0.003-0.3 mM for Spm and 0.01-0.4 mM for Spmd, while with the SMO biosensor, a linear range of 0.004-0.5 mM for Spm has been obtained. The main kinetic parameters apparent Michaelis constant (K M), turnover number (K cat) and steady-state current (I max) were determined. The proposed device was then applied to the determination of biogenic amines in blood samples. The results obtained were in good agreement with those obtained with the GC-MS reference method.

Taibaiella yonginensis sp. nov., a bacterium isolated from soil of Yongin city.

A novel bacterial strain, designated as THG-SC4(T), was isolated from a soil sample collected from Yongin city in South Korea. Cells of the strain were Gram-negative, aerobic, rod-shaped and non-motile. The strain grew optimally at 28-30 °C; at pH 7.0 and in the absence of NaCl. Flexirubin-type pigments were found to be present. On the basis of 16S rRNA gene sequence similarity, strain THG-SC4(T) was shown to belong to the genus Taibaiella and shares high sequence similarity with Taibaiella smilacinae KCTC 32316(T) (95.4 %), followed by Taibaiella koreensis KACC 17171(T) (94.3 %) and Taibaiella chishuiensis JCM 19637(T) (94.2 %). The DNA G+C content of the novel isolate was determined to be 43.1 mol% and MK-7 was identified as the predominant isoprenoid quinone. The only polyamine was homospermidine. The major polar lipids were phosphatidylethanolamine, an unidentified aminophospholipid and an unidentified polar lipid. The predominant fatty acids were identified as iso-C15:0, iso-C15:1 G, C16:0 and iso-C17:03-OH. On the basis of data from this polyphasic taxonomic study, strain THG-SC4(T) is considered to represent a novel species of the genus Taibaiella, for which the name Taibaiella yonginensis sp. nov. is proposed. The type strain is THG-SC4(T) (=KACC 18372(T) = CCTCC AB 2014316(T)).

Sphingomonas panacis sp. nov., isolated from rhizosphere of rusty ginseng.

The type strain DCY99(T) was isolated from soil collected from a ginseng field in Hwacheon, Republic of Korea. Strain DCY99(T) is Gram-negative, non-spore forming, motile, rod-shaped, and strictly aerobic. The bacteria grow optimally at 25-30 °C and pH 6.0-6.5. Phylogenetically, strain DCY99(T) is most closely related to Sphingomonas oligophenolica JCM 12082(T), followed by Sphingomonas asaccharolytica KCTC 2825(T), Sphingomonas mali KCTC 2826(T), Sphingomonas cynarae JCM17498(T), Sphingomonas pruni KCTC 2824(T), and Sphingomonas glacialis DSM 22294(T). The DNA-DNA relatedness between strain DCY99(T) and S. oligophenolica JCM 12082(T) was 15.6 ± 0.4 %, and the DNA G+C content of strain DCY99(T) was 64.4 mol%. An isoprenoid quinone was detected and identified as ubiquinone Q-10, and sym-homospermidine was identified as the major polyamine of DCY99(T). The major polar lipids were identified as sphingoglycolipid, diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, and phosphatidylcholine. C14:02OH, C16:0, and summed feature 8 (C18:1 ω7c:/C18:1 ω6c) were identified as the major fatty acids present in DCY99(T). The results of physiological and biochemical tests allowed strain DCY99(T) to be differentiated phenotypically from other recognized species belonging to the genus Sphingomonas. Therefore, it is suggested that the newly isolated organism represents a novel species, for which the name Sphingomonas panacis sp. nov. is proposed with the type strain designated as DCY99(T) (=JCM 30806(T) =KCTC 42347(T)).

Sphingomonas aeria sp. nov. from indoor air of a pharmaceutical environment.

A proteobacterial strain designated R1-3(T) was isolated from indoor air of a pharmaceutical environment. Cells were Gram-stain-negative, oxidase- and catalase-positive, aerobic, motile and rod-shaped. Strain R1-3(T) grew optimally at pH 7, 30 °C and in 0-2 % NaCl on R2A agar. The 16S rRNA gene sequence analysis indicated that strain R1-3(T) belongs to the genus Sphingomonas, and is closely related to Sphingomonas paucimobilis ATCC 29837(T) (98.4 % sequence similarity). However, the DNA-DNA relatedness between the two strains was 43 ± 5 % (reciprocal = 37 ± 3 %), which was well below the suggested level for species distinction. Sphingomonas yabuuchiae GTC868(T) (97.7 % 16S rRNA gene sequence similarity) and Sphingomonas pseudosanguinis G1-2(T) (97.6 %) were also found as distantly related taxa. Strain R1-3(T) was sensitive to most of the tested antibiotics except for erythromycin and streptomycin. The major fatty acid was a summed feature consisting of C18:1 ω7c and/or C18:1 ω6c, and minor proportions of C14:0 2-OH, C16:0 and a summed feature consisting of C16:1 ω7c and/or C16:1 ω6c were also present. The DNA G + C content was 67.2 ± 1.0 mol%. The major polyamines were sym-homospermidine and spermidine. The major polar lipids were phosphatidylglycerol, phosphatidylethanolamine, diphosphatidylglycerol, and minor amounts of a sphingoglycolipid, a phospholipid, an aminoglycolipid and an unidentified lipid were also present. The phenotypic, phylogenetic and chemotaxonomic data not only supported the affiliation of strain R1-3(T) to the genus Sphingomonas, but also distinguished R1-3(T) from related species. On the basis of physiological, chemotaxonomic and phylogenetic evidences, strain R1-3(T) clearly merits recognition as a novel species of Sphingomonas, for which the name Sphingomonas aeria sp. nov. is proposed. The type strain is R1-3(T) (= KCTC 42061(T) = JCM 19859(T)).

Sphingomonas chloroacetimidivorans sp. nov., a chloroacetamide herbicide-degrading bacterium isolated from activated sludge.

Strain Y1(T), a Gram-negative, non-spore-forming, rod-shaped bacterium, was isolated from activated sludge. This strain is able to degrade several commonly used chloroacetamide herbicides, such as butachlor, acetochlor and alachlor. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain Y1(T) is a member of the genus Sphingomonas and shows high sequence similarities with S. starnbergensis 382(T) (95.7 %), S. sanxanigenens NX02(T) (95.7 %) and S. haloaromaticamans A175(T) (95.3 %), and shows low (<95 %) sequence similarities to all other Sphingomonas species. Chemotaxonomic analysis revealed that strain Y1(T) possesses Q-10 as the predominant ubiquinone, C14:0 2-OH as the major 2-hydroxy fatty acid and sym-homospermidine as the major polyamine. The main cellular fatty acids of strain Y1(T) were found to be C18:1 ω7c (38.2 %), C16:1 ω6c/C16:1 ω7c (28.5 %), C16: 0 (10.7 %) and C14:0 2-OH (14.3 %). The main polar lipids were determined to be diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, sphingoglycolipids (SGL1-SGL3), phosphatidyl dimethylethanolamine and aminophospholipid. The DNA G+C content was found to be 66 ± 0.4 mol%. Based on phylogenetic analysis, phenotypic characteristics and chemotaxonomic data, strain Y1(T) is considered to represent a novel species of the genus Sphingomonas, for which the name Sphingomonas chloroacetimidivorans sp. nov. is proposed. The type strain is Y1(T) (=CCTCC AB 2011178(T) = KACC 16607(T)).