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Soils are the largest reservoir of microplastics (MPs) on earth. Since MPs can remain in soils for a very long time, their effects are magnified. In this study, different concentrations of polyethylene (PE) MPs derived from commercial resins (0%, 1%, 7%, and 14%, represented as MP_0, MP_1, MP_7, and MP_14) were added to soils to assess the changes in the soils' chemical properties, enzyme activities, and bacterial communities during a 70-day incubation period. The results show that PE MP treatments with low concentrations differed from other treatments in terms of exchangeable Ca and Mg, whereas at high concentrations, the pH and availability of phosphate ions differed. Fluorescein diacetate (FDA), acid phosphatase (ACP), and N-acetyl-β-d-glucosaminidase (NAG) enzyme activities exhibited a dose-related trend with the addition of the PE MPs; however, the average FDA and ACP activities were significantly affected only by MP_14. Changes in the microbial communities were observed at both the phylum and family levels with all PE MP treatments. It was revealed that even a low dosage of PE MPs in soils can affect the functional microbes, and a greater impact is observed on those that can survive in polluted environments with limited resources.

Background: Lumbosacral muscle strain (LMS) is common in Chinese elite trampoline athletes. Advanced lumbar muscle activation is necessary for postural control before upper extremity voluntary movements, called anticipatory postural adjustment to reduce internal postural interference (IPI). The potential of delayed lumbar muscle activation has been reported in patients with non-specific LBP (NLBP) in response to IPI. However, it remains unknown whether this effect exists in elite trampoline athletes. There is also limited literature reporting the rehabilitation of LMS in this population. This study first aimed to explore whether elite trampoline athletes with LMS experience delayed activation of lumbar muscles under IPI. The secondary aim was to preliminarily evaluate an integrative rehabilitation program's effectiveness., Materials and Methods: Ten elite trampoline athletes with LMS were recruited and received 10 sessions of integrative rehabilitation, including extracorporeal shock wave therapy, acupuncture, Tui-na, and spine function exercises. At baseline and after all sessions, the relative activation time of the lumbar muscles under IPI in a modified rapid arm-rise test was used as a primary outcome measure. The secondary measures included a visual analog scale (VAS) and a questionnaire to assess low back pain (LBP) and athletic training performance., Results: The relative activation time of the lumbar muscles under IPI was delayed at baseline, but significantly decreased after the intervention ( P  < 0.05). The VAS was significantly decreased after the intervention ( P  < 0.05). There was no significant correlation between the difference in VAS and in activation time of the lumbar muscles before and after the intervention ( P  > 0.05)., Conclusions: Elite trampoline athletes with LMS had delayed activation in their lumbar muscles under IPI. Integrative rehabilitation was effective in LBP relief and neuromuscular control of the lumbar muscles, and impacted positively on training performance. Future studies with a larger sample size, a control group, and long-term follow-ups are needed to further examine the efficacy of integrative rehabilitation in elite trampoline athletes with LMS. Additionally, the application of this approach in athletes with LMS or LBP in other sports, particularly those involving IPI, should be explored., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (© 2024 Ma, Wu, Zhang, Zhao, Shen, Feng, Zhang, Zhu and Xu.)

Several bacteria of environmental and clinical origins, including some human-associated strains secrete a cross-kingdom signaling molecule indole-3-acetic acid (IAA). IAA is a tryptophan (trp) derivative mainly known for regulating plant growth and development as a hormone. However, the nutritional sources that boost IAA secretion in bacteria and the impact of secreted IAA on non-plant eukaryotic hosts remained less explored. Here, we demonstrate significant trp-dependent IAA production in Pseudomonas juntendi NEEL19 when provided with ethanol as a carbon source in liquid cultures. IAA was further characterized to modulate the odor discrimination, motility and survivability in Drosophila melanogaster. A detailed analysis of IAA-fed fly brain proteome using high-resolution mass spectrometry showed significant (fold change, ± 2; p ≤ 0.05) alteration in the proteins governing neuromuscular features, audio-visual perception and energy metabolism as compared to IAA-unfed controls. Sex-wise variations in differentially regulated proteins were witnessed despite having similar visible changes in chemo perception and psychomotor responses in IAA-fed flies. This study not only revealed ethanol-specific enhancement in trp-dependent IAA production in P. juntendi, but also showed marked behavioral alterations in flies for which variations in an array of proteins governing odor discrimination, psychomotor responses, and energy metabolism are held responsible. Our study provided novel insights into disruptive attributes of bacterial IAA that can potentially influence the eukaryotic gut-brain axis having broad environmental and clinical implications., (© 2024. The Author(s).)

In Taiwan, the pesticides dimethomorph and imidacloprid are recommended for pest control in vineyards. Therefore, tank-mixing of these two pesticides is usually a routine practice before application. This study analyzed the influence of vineyard soil microbial flora under the recommended and high dosages (100 times the recommended dosage) of dimethomorph and imidacloprid. Individual and combined applications of pesticides were also tested through batches of soil incubation experiments. Four treatments-control (C), dimethomorph (DT), imidacloprid (IM), and mixed application of dimethomorph and imidacloprid (ID)-were used in the experimental design. From the soil metabolism, no significant reaction was observed after 2 months in the recommended dosage group, regardless of whether the pesticides were being applied individually or combined. For the high dosage, imidacloprid showed a higher effect than the co-exposure treatments, showing a possible prolonged effect after its repetitive application. From PCoA analysis, pesticide treatments altered the soil ecology after 2 months, and the effect of imidacloprid can be explicitly observed at high dosages. At the phylum level, Acidobacteria can indicate pesticide application around the recommended dosage. It was inhibited by ID on day 7 and was augmented by all pesticides on day 63. The effect of the recommended dosage of pesticide mixtures after 2 months of incubation was revealed in the minor families Gemmataceae and Pirellulaceae , while the high dosage treatments affected both the core and the minor families. Our findings verified the changes in the composition of microbial communities upon pesticide application, which would affect carbon, nitrogen, sulfur, phosphorous cycles, and contaminant removal ability within the vineyard., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Chang, Shen, Lai, Liao and Chen.)

The aim of this study was to develop an efficient bioinoculant for amelioration of adverse effects from chilling stress (10°C), which are frequently occurred during rice seedling stage. Seed germination bioassay under chilling condition with rice (Oryza sativa L.) cv. Tainan 11 was performed to screen for plant growth-promoting (PGP) bacteria among 41 chilling-tolerant rice endophytes. And several agronomic traits were used to evaluate the effects of bacterial inoculation on rice seedling, which were experienced for 7-d chilling stress in walk-in growth chamber. The field trials were further used to verify the performance of potential PGP endophytes on rice growth. A total of three endophytes with multiple PGP traits were obtained. It was demonstrated that Pseudomonas sp. CC-LS37 inoculation led to 18% increase of maximal efficiency of Photosystem II (PSII) after 7-d chilling stress and 7% increase of chlorophyll a content, and 64% decline of malondialdehyde content in shoot after 10-d recovery at normal temperature in walk-in growth chamber. In field trial, biopriming of seeds with strain CC-LS37 caused rice plants to increase shoot chlorophyll soil plant analysis development values (by 2.9% and 2.5%, respectively) and tiller number (both by 61%) under natural climate and chilling stress during the end of tillering stage, afterward 30% more grain yield was achieved. In conclusion, strain CC-LS37 exerted its function in increase of tiller number of chilling stress-treated rice seedlings via improvement of photosynthetic characteristics, which in turn increases the rice grain yield. This study also proposed multiple indices used in the screening of potential endophytes for conferring chilling tolerance of rice plants., (© The Author(s) 2022. Published by Oxford University Press on behalf of Applied Microbiology International.)

Tea leaves possess numerous volatile organic compounds (VOC) that contribute to tea's characteristic aroma. Some components of tea VOC were known to exhibit antimicrobial activity; however, their impact on bacteria remains elusive. Here, we showed that the VOC of fresh aqueous tea leaf extract, recovered through hydrodistillation, promoted cell division and tryptophan-dependent indole-3-acetic acid (IAA) production in Pseudomonas sp. NEEL19, a solvent-tolerant isolate of the tea phylloplane. 1-octanol was identified as one of the responsible volatiles stimulating cell division, metabolic change, swimming motility, putative pili/nanowire formation and IAA production, through gas chromatography-mass spectrometry, microscopy and partition petri dish culture analyses. The bacterial metabolic responses including IAA production increased under 1-octanol vapor in a dose-dependent manner, whereas direct-contact in liquid culture failed to elicit such response. Thus, volatile 1-octanol emitting from tea leaves is a potential modulator of cell division, colonization and phytohormone production in NEEL19, possibly influencing the tea aroma.

Plant growth-promoting bacteria (PGPB) are beneficial microbes that increase plant growth and yield. However, limited information is currently available on PGPB in onion (Allium cepa Linn.). The aims of the present study were to isolate and identify PGPB in onion and examine the effects of isolated PGPB on germination and growth during the vegetative stage in onion, pak choy (Brassica chinensis), and sweet pepper (Capsicum annuum). Twenty-three strains of PGPB were isolated from the roots, bulbs, and rhizosphere soil of onion. All isolated bacterial strains showed one or more PGP traits, including indole acetic acid production, phosphate solubilization ability, and 1-aminocyclopropane-1-carboxylate deaminase and nitrogenase activities; most of these traits were derived from Bacillus sp., Microbacterium sp., and Pseudomonas sp. Eight bacteria that exhibited strong abilities to produce indole acetic acid were selected for a Petri dish trial, soil pot test, and vermiculate pot test. The Petri dish trial showed that strains ORE8 and ORTB2 simultaneously increased radicle and hypocotyl lengths in onion, but inhibited growth in sweet pepper after 7 d. The soil pot experiment on onion revealed that strains ORE5, ORE8, and ORTB2 strongly promoted growth during the vegetative stage with only a half dose of chemical fertilizer. The present results indicate that ORE8 (Bacillus megaterium) and ORTB2 (Pantoea sp.) are the most promising biofertilizers of onion and may simultaneously inhibit the seedling growth of other plants.

The present study was performed to investigate root-associated bacteria from Platycodon grandiflorum, a medicinal plant commonly grown in East Asia. Isolates were obtained from the rhizosphere or root interior with various culture media, and phylogenetic analyses were performed based on their 16S rDNA sequences. In consideration of practical applications, traits related to plant growth promotion and niche adaptation were assessed in several endophytic strains with fewer biosafety concerns. The effects of a bacterial inoculation on seedling and mature plant growth were evaluated. Seventeen genera that encompassed more than 30 bacterial lineages were successfully retrieved from the roots, the majority of which have not been reported as P. grandiflorum-associated bacteria, particularly for non-negligible Proteobacteria. Although nitrogen-fixing or phosphate-solubilizing and indole acetic acid-producing activities were recorded in all of the strains selected, these strains were beneficial or detrimental to plant growth as evidenced by their influence on the length of seedlings and biomass of mature plants. Among the 4 endophytic Rhizobium species tested in the present study, the potentially novel Rhizobium sp. BF-E16, which was more compatible with the non-leguminous medicinal plant P. grandiflorum, was identified. Other than plant growth-promoting traits, characteristics such as plant constituent-hydrolyzing activities need to be taken into consideration and their roles clarified when investigating plant growth-promoting rhizobacteria.

A novel yellow-pigmented, aerobic, rod-shaped, non-motile bacterium, designated strain CCMF41T, was isolated from rhizosphere soil of maize (Zea mays) collected in Wufeng District, Taichung, Taiwan. Strain CC-MF41T exhibited 16S rRNA gene sequence similarity of 97.5, 97.3, 97.2 and 97.1% to Leucobacter chironomi MM2LBT (and ‘Leucobacter kyeonggiensis’F3-P9 and ‘L. humi’ Re-6, the names of which have not been validly published), Leucobactertardus K70/01T, L. komagatae IFO 15245T and ‘Leucobacter margaritiformis’ A23. However,CC-MF41T and ‘L. margaritiformis’ A23 formed a loosely bound phylogenetic lineage (with alow bootstrap value) associated with species of the genus Leucobacter. In DNA–DNA reassociation experiments, the relatedness of strain CC-MF41T to L. chironomi DSM 19883T was 57.1% (reciprocal value 29.1 %). The DNA G+C content of strain CC-MF41T was 72.1 mol% and the cell-wall peptidoglycan contained 2,4-diaminobutyric acid, alanine, glycine,glutamic acid and threonine. The major menaquinone was MK-11 and the predominant fatty acids were iso-C16 : 0, anteiso-C15 : 0 and anteiso-C17 : 0. The polar lipid profile of strain CCMF41T contained major amounts of diphosphatidylglycerol followed by an unidentified glycolipid, phosphatidylglycerol and an unknown phospholipid. Based on its phylogenetic,phenotypic and chemotaxonomic distinctiveness, strain CC-MF41T represents a novel species of Leucobacter, for which the name Leucobacter zeae sp. nov. is proposed. The type strain isCC-MF41T (=BCRC 80515T=LMG 27265T).

An aerobic, Gram-stain-negative, rod or spiral-shaped diazotrophic bacterium (designated strain CC-LY788T), was isolated from agricultural soil in Taiwan. Strain CC-LY788T was able to grow at 25-40 °C, pH 6.0-8.0 and tolerated NaCl to 2.0% (w/v). Positive for nitrogen fixation with the activity recorded as 6.5 nmol ethylene h(-1). Strain CC-LY788T showed highest 16S rRNA gene sequence similarity to Azospirillum picis DSM 19922T (97.2%) and Azospirillum rugosum DSM 19657T (97.1%) and lower sequence similarities (<96.6%) to all other species of the genus Azospirillum. According to the DNA-DNA hybridization, the relatedness values of strain CC-LY788T with A. picis DSM 19922T and A. rugosum DSM 19657T were 51.1±5.5% and 46.8±2.1%, respectively. Strain CC-LY788T was positive for the rapid identification of the genus-specific primer set. The respiratory quinone system was ubiquinone (Q-10) and the DNAG+C content was 69.8 mol%. The major fatty acids found in strain CC-LY788T were C16 : 0, C18 : 1 2-OH, C14 : 0 3-OH/C16 : 1 iso I (summed feature 2), C16 : 1ω7c/C16 : 1ω6c (summed feature 3), C18 : 0 ante/C18 : 2ω6,9c (summed feature 5) and C18 : 1ω7c/C18 : 1ω6c (summed feature 8). Based on the phylogenetic, phenotypic and chemotaxonomic features, strain CC-LY788T represents a novel species of the genus Azospirillum, for which the name Azospirillum soli sp. nov. is proposed. The type strain is CC-LY788T (=BCRC 80569T=JCM 18820T).

A Gram-stain-negative, aerobic, non-motile, rod-shaped, flexirubin-producing bacterium, designated strain CC-CZW010T, was isolated from the edible sea urchin Tripneustes gratilla in Penghu Island, Taiwan. The isolate grew optimally at pH 7.0 and 30 °C in the presence of 2 % (w/v) NaCl. The most closely related strains in terms of 16S rRNA gene sequence similarity were Chryseobacterium taihuense NBRC 108747T (97.6 %) and Chryseobacterium aquaticum KCTC 12483T (96.7 %). Phylogenetic analyses based on 16S rRNA gene sequences revealed a distinct taxonomic position attained by strain CC-CZW010T with respect to other species of the genus Chryseobacterium. Strain CC-CZW010T possessed iso-C15 : 0, anteiso-C15 : 0, iso-C17 :  0 3-OH, summed feature 3 (comprising C16 : 1ω7c/C16 : 1ω6c) and summed feature 9 (comprising C16 :  0 10-methyl/iso-C17 : 1ω9c) as predominant fatty acids. The major polar lipid profile consisted of phosphatidylethanolamine, two unidentified lipids and five aminolipids. The polyamine pattern contained the major compound sym-homospermidine. Menaquinone 6 (MK-6) was the predominant respiratory quinone, and the G+C content of the genomic DNA was 36.4 mol%. According to distinct phylogenetic, phenotypic and chemotaxonomic features, strain CC-CZW010T represents a novel species of the genus Chryseobacterium, for which the name Chryseobacterium echinoideorum sp. nov. is proposed. The type strain is CC-CZW010T ( = BCRC 80786T = JCM 30470T).

A Gram-stain-variable, short-rod-shaped, endospore-forming, strictly aerobic, non-motile, chitinolytic and endophytic bacterium, designated strain CC-Alfalfa-19T, exhibiting unusual bipolar appendages was isolated from a root nodule of alfalfa (Medicago sativa L.) in Taiwan and subjected to a polyphasic taxonomic study. Based on 16S rRNA gene sequence analysis, strain CC-Alfalfa-19T was found to be most closely related to Paenibacillus puldeungensis CAU 9324T (95.2 %), whereas other species of the genus Paenibacillus shared ≤ 95.0 % sequence similarity. The phylogenetic analysis revealed a distinct phyletic lineage established by strain CC-Alfalfa-19T with respect to other species of the genus Paenibacillus. Fatty acids comprised predominantly anteiso-C15 : 0, C16 : 0, anteiso-C17 : 0 and iso-C16 : 0. Menaquinone 7 (MK-7) was identified as the sole respiratory quinone and the genomic DNA G+C content was 42.7 mol%. Polar lipids included diphosphatidylglycerol, phosphatidylglycerol, phosphatidylmonomethylethanolamine, phosphatidylethanolamine, an unidentified glycolipid and an unidentified lipid. The diagnostic diamino acid found in the cell-wall peptidoglycan was meso-diaminopimelic acid. Based on the polyphasic taxonomic evidence that was in line with the genus Paenibacillus and additional distinguishing characteristics, strain CC-Alfalfa-19T is considered to represent a novel species, for which the name Paenibacillus medicaginis sp. nov. (type strain CC-Alfalfa-19T = BCRC 80441T = JCM 18446T) is proposed.

A Gram-stain-negative, aerobic, rod-shaped, yellow-pigment-producing bacterium (designated strain CC-CZW007(T)) was isolated from seafood samples (sea urchins) at Penghu Island in Taiwan. Strain CC-CZW007(T) grew optimally at pH 7.0 and 30 °C in the presence of 3% (w/v) NaCl. The novel strain shared highest 16S rRNA gene sequence similarity to Vitellibacter vladivostokensis JCM 11732(T) (96.8%), Vitellibacter soesokkakensis KCTC 32536(T) (96.4%), Vitellibacter nionensis KCTC 32420(T) (95.8%) and Vitellibacter aestuarii JCM 15496(T) (95.6%) and lower sequence similarity to members of other genera. Phylogenetic analyses based on 16S rRNA genes revealed a distinct taxonomic position attained by strain CC-CZW007(T) with respect to other species of the genus Vitellibacter. The major fatty acids were iso-C15 : 0 and iso-C17 : 0 3-OH. The polar lipid profile was composed of major amounts of phosphatidylethanolamine, unidentified lipids and aminolipids; a moderate amount of aminophospholipid was also detected. The DNA G + C content was 34.7 mol%. The predominant quinone system was menaquinone (MK-6). On the basis of polyphasic taxonomic evidence presented here, strain CC-CZW007(T) is proposed to represent a novel species within the genus Vitellibacter, for which the name Vitellibacter echinoideorum sp. nov. is proposed. The type strain is CC-CZW007(T) ( = BCRC 80886(T) = JCM 30378(T)).

A Gram-stain-positive, aerobic, non-motile, rod-shaped, yellow-pigment-producing bacterium, (designated strain CC-CZW004T), was isolated from seafood samples (sea urchins) at Penghu Island in Taiwan. Strain CC-CZW004T grew optimally at pH 7.0 and 30 °C in the presence of 3 % (w/v) NaCl. Phylogenetic analyses based on 16S rRNA genes revealed a distinct taxonomic position attained by strain CC-CZW004T with respect to other members of the genus Nocardioides. The novel strain shared highest 16S rRNA gene sequence similarities to Nocardioides daejeonensis JCM 16922T (96.4 %), Nocardioides pacificus JCM 19260T (96.3 %), and Marmoricola scoriae JCM 17444T (96.2 %). The major fatty acids of strain CC-CZW004T consisted of C17:0, C16:1ω5c, C17:1ω8c`, iso-C16:0 and C19:1ω11c/C19:1ω9c (summed feature 6). The diagnostic diamino acid in the cell wall was ll-2,6-diaminopimelic acid. The polar lipid profile was composed of major amounts of phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol and aminophospholipid. The DNA G+C content was 73.3 mol%. The predominant quinone system was menaquinone (MK-8). On the basis of polyphasic taxonomic evidences presented here, strain CC-CZW004T represents a novel species of the genus Nocardioides, for which the name Nocardioides echinoideorum sp. nov. is proposed. The type strain is CC-CZW004T ( = BCRC 16974T = JCM 30276T).

Discharging untreated highly acidic (pH<4.0), organic and nutrients rich monosodium glutamate wastewater (MW), and highly alkaline (pH>10.0) paper-mill wastewater (PW) causes environmental pollution. When acidity of MW neutralized (pH 6.5±0.1) with PW and lime (treatments represented as MW+PW and MW+Lime), then MW may be utilized as a potential source of nutrients and organic carbon for sustainable food production. Objectives of this study were to compare the effects of PW and lime neutralized MW and chemical fertilizers on maize (Zea mays L. cv. Snow Jean) plant growth, yield, nutrients uptake, soil organic matter and humic substances. The field experiment was carried out on maize using MW at 6000 L ha(-1). Impacts of the MW application on maize crop and soil properties were evaluated at different stages. At harvest, plant height, and plant N and K uptake were higher in MW treatment. Leaf area index at 60 days after sowing, plant dry matter accumulation at harvest, and kernels ear(-1) and 100-kernel weight were higher in MW+Lime treatment. Kernel N, P, K, Mn, Fe and Zn, and plant Zn uptake were highest in MW+Lime. Plant Fe uptake, and soil organic matter and humic substances were highest in MW+PW. The MW+PW and MW+Lime treatments exhibited comparable results with chemically fertilized treatment. The MW acidity neutralized with lime showed positive impacts on growth, yield and nutrients uptake; nevertheless, when MW pH neutralized with PW has an additional benefit on increase in soil organic matter and humic substances., (Copyright © 2014. Published by Elsevier Ltd.)

A taxonomic study was carried out on a novel aerobic bacterial strain (designated CC-LY736(T)) isolated from a fermentor in Taiwan. Cells of strain CC-LY736(T) were Gram-stain negative, spiral-shaped and motile by means of a monopolar flagellum. Strain CC-LY736(T) shared the greatest degree of 16S rRNA gene sequence similarity to Azospirillum irakense DSM 11586(T) (97.2 %), Rhodocista centenaria JCM 21060(T) (96.3 %) and Rhodocista pekingensis JCM 11669(T) (96.1 %). The major fatty acids were C16:0, C16:1 ω5c, C19:0 cyclo ω8c, C18:1 ω7c/C18:1 ω6c, C16:0 3-OH and C18:1 2-OH. The predominant polar lipids included phosphatidylcholine, phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylmethylethanolamine, phosphatidyldimethylethanolamine and two unidentified glycolipids. The common major respiratory quinone was ubiquinone Q-10 and predominant polyamines were sym-homospermidine and putrescine. The DNA G+C content of strain CC-LY736(T) was 67.6 ± 0.1 mol %. During phylogenetic analysis, strain CC-LY736(T) formed a unique phyletic lineage associated with Rhodocista species. However, the combination of genetic, chemotaxonomic and physiological data clearly indicated that strain CC-LY736(T) was a novel representative of the family Rhodospirillaceae. Based on the polyphasic comparison, the name Niveispirillum fermenti gen. nov., sp. nov. is proposed; the type strain of the type species is CC-LY736(T) (= BCRC 80504(T) = LMG 27263(T)). In addition, the reclassifications of Azospirillum irakense as Niveispirillum irakense comb. nov. (type strain KBC1(T) = ATCC 51182(T) = BCRC 15764(T) = CIP 103311(T)), and Azospirillum amazonense as Nitrospirillum amazonense gen. nov., sp. nov. (type strain Am14(T) = ATCC 35119(T) = BCRC 14279(T) = DSM 3787(T)) are proposed based on the polyphasic taxonomic data obtained in this study.

An aerobic, Gram-stain-negative, rod-shaped bacterium (designated strain CC-G9A(T)), motile by a polar-flagellum, was isolated from a hot spring water sample in Taiwan. Strain CC-G9A(T) could grow at 20-42 °C, pH 6.0-10.0 and tolerate up to 7% (w/v) NaCl. The 16S rRNA gene sequence analysis of strain CC-G9A(T) showed pairwise sequence similarity to Pseudomonas mendocina LMG 1223(T) (97.7%), Pseudomonas alcaligenes ATCC 14909(T) (97.8 %), Pseudomonas alcaliphila DSM 17744(T) (97.8 %), Pseudomonas toyotomiensis JCM 15604(T) (97.6 %), Pseudomonas oleovorans subsp. lubricantis DSM 21016(T) (97.6 %) and Pseudomonas argentinensis BCRC 17807(T) (97.5 %), and lower sequence similarity to other species of the genus Pseudomonas. According to DNA-DNA association analysis, the relatedness of strain CC-G9A(T) to P. mendocina BCRC 10458(T), P. alcaliphila DSM 17744(T), P. alcaligenes BCRC 11893(T), P. oleovorans subsp. lubricantis DSM 21016(T), P. argentinensis BCRC 17807(T) and P. oleovorans subsp. oleovorans BCRC 11902 was 55.1±3.1, 13.7±1.5, 14.1±1.8, 58.5±1.1, 28.9±2.0 and 28.6±1.8 %, respectively. The evolutionary trees reconstructed based on 16S rRNA, gyrB and rpoB gene sequences revealed varying phylogenetic neighbourhoods of strain CC-G9A(T) with regard to the most closely related type strains. The predominant quinone system was ubiquinone (Q-9) and the DNA G+C content was 64.3±1.3 mol%. The major fatty acids were C10 : 0 3-OH, C12 : 0, C12 : 0 3-OH, C16 : 0 and summed features 3 and 8 consisting of C16 : 1ω7c/C16 : 1ω6c and C18 : 1ω7c/C18 : 1ω6c, respectively. The major polar lipids were diphosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine and phosphatidylglycerol. According to distinct phylogenetic, phenotypic and chemotaxonomic features, strain CC-G9A(T) is proposed to represent a novel species within the genus Pseudomonas for which the name Pseudomonas guguanensis sp. nov. is proposed. The type strain is CC-G9A(T) ( = BCRC 80438(T) = JCM 18416(T)).

An aerobic, Gram-stain-negative, spiral or rod-shaped, non-spore-forming, diazotrophic bacterium (strain CC-LY743(T)) was isolated from a fermentative tank in Taiwan. Strain CC-LY743(T) was able to grow at 20-37 °C and pH 6.0-8.0 and tolerated up to 3.0 % (w/v) NaCl. It was positive for nitrogen fixation, with activity of 10.6 nmol ethylene h(-1). 16S rRNA gene sequence analysis of strain CC-LY743(T) showed highest similarity to Azospirillum picis DSM 19922(T) (96.1 %), Azospirillum oryzae JCM 21588(T) (96.0 %) and Azospirillum rugosum DSM 19657(T) (96.0 %) and lower similarity (<96.0 %) to all other Azospirillum species. Highest nifH gene sequence similarities were obtained with Azospirillum brasilense BCRC 12270(T) (92.0 %), Azospirillum formosense BCRC 80273(T) (92.3 %) and A. rugosum DSM 19657(T) (91.8 %). It was positive in the rapid identification by a genus-specific primer set. The predominant quinone system was ubiquinone 10 (Q-10) and the DNA G+C content was 69.6±0.1 mol%. The major fatty acids found in strain CC-LY743(T) were n-C16 : 0, C19 : 0 cyclo ω8c, C14 : 0 3-OH/C16 : 1 iso I, C16 : 1ω7c/C16 : 1ω6c and C18 : 1ω7c/C18 : 1ω6c. Based on its phylogenetic, phenotypic and chemotaxonomic features, strain CC-LY743(T) is considered to represent a novel species within the genus Azospirillum for which the name Azospirillum fermentarium sp. nov. is proposed. The type strain is CC-LY743(T) ( = BCRC 80505(T) = JCM 18688(T) = LMG 27264(T)).

A novel Gram-reaction-negative, yellow-pigmented, aerobic, non-motile and rod-shaped bacterium designated strain CC-YHH031(T) was isolated from an agricultural soil collected at Chiayi County, Taiwan. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain CC-YHH031(T) formed a discrete monophyletic lineage in the family Xanthomonadaceae, sharing high pairwise sequence similarity of 93.5-95.2 and 94.8% with species of the genus Dokdonella (94.9% similarity to the type strain of the type species) and Aquimonas voraii GPTSA 20(T), respectively. The genomic DNA G+C content of strain CC-YHH031(T) was 68.6 ± 0.7 mol% and the predominant respiratory quinone was ubiquinone Q-8. Spermidine was the principal polyamine, with minor amounts of putrescine. Major fatty acids (>5% of total fatty acids) were iso-C(16:00, iso-C(15:0), C(16:1)ω7c and/or C(16:1)ω6c (summed feature 3), iso-C(17:1)ω9c, iso-C(14:0), iso-C(11:0) and iso-C(11:0) 3-OH. The polar lipid profile of strain CC-YHH031(T) included phosphatidylethanolamine, phosphatidylmonomethylethanolamine, diphosphatidylglycerol, phosphatidylglycerol, two unidentified aminophospholipids (APL1-2) and four unidentified phospholipids (PL1-4). Strain CC-YHH031(T) was distinguished particularly from the type species of the genus Dokdonella (Dokdonella koreensis) by the presence of major amounts of iso-C(14:0) and summed feature 3 and minor amounts of iso-C(17:0) and by the complete absence of anteiso-C(17:0), the presence of PL1-3 and APL1-2, the absence of APL3 and the presence of putrescine in the former. On the basis of distinguishing genotypic and phenotypic evidence, strain CC-YHH031(T) is proposed to represent a novel genus and species within the family Xanthomonadaceae, for which the name Chiayiivirga flava gen. nov., sp. nov. is proposed. The type strain of Chiayiivirga flava is CC-YHH031(T) ( =BCRC 80274(T) =DSM 24163(T)).

An aerobic, Gram-stain-negative, rod-shaped bacterium with a single polar flagellum, designated CC-OPY-1(T), was isolated from an oil-contaminated site in Taiwan. CC-OPY-1(T) produces siderophores, and can grow at temperatures of 25-37 °C and pH 5.0-9.0 and tolerate <5 % (w/v) NaCl. The 16S rRNA gene sequence analysis of CC-OPY-1(T) showed high pairwise sequence similarity to Pseudomonas alcaligenes BCRC 11893(T) (97.1 %), Pseudomonas. alcaliphila DSM 17744(T) (97.1 %), Pseudomonas tuomuerensis JCM 14085(T) (97.1 %), Pseudomonas toyotomiensis JCM 15604(T) (96.9 %) and lower sequence similarity to remaining species of the genus Pseudomonas. The phylogenetic trees reconstructed based on gyrB and rpoB gene sequences supported the classification of CC-OPY-1(T) as a novel member of the genus Pseudomonas. The predominant quinone system of strain CC-OPY-1T was ubiquinone (Q-9) and the DNA G+C content was 68.4 ± 0.3 mol%. The major fatty acids were C12 : 0, C16 : 0, C17 : 0 cyclo and summed features 3 and 8 consisting of C16 : 1ω7c/C16 : 1ω6c and C18 : 1ω7c/C18 : 1ω6c, respectively. The major polar lipids were phosphatidylethanolamine (PE), phosphatidylglycerol (PG), diphosphatidylglycerol (DPG), phosphatidylcholine (PC) and two unknown phospholipids (PL1-2). Due to distinct phylogenetic, phenotypic and chemotaxonomic features, CC-OPY-1(T) is proposed to represent a novel species within the genus Pseudomonas for which the name Pseudomonas sagittaria sp. nov. is proposed. The type strain is CC-OPY-1(T) ( = BCRC 80399(T) = JCM 18195(T)).

Two novel siderophore-producing strains designated CC-CFT023(T) and CC-CFT034(T), isolated from a rusty iron plate, are characterized in this study. Cells of both strains were catalase- and oxidase-positive. Strain CC-CFT023(T) showed a high pairwise 16S rRNA gene sequence similarity to Aureimonas altamirensis LMG 23375(T) (96.0 %), whereas, strain CC-CFT034(T) shared a maximum similarity to Aureimonas ureilytica DSM 18598(T) (95.8 %). The predominant quinone system was ubiquinone (Q-10) and the DNA G+C contents of CC-CFT023(T) and CC-CFT034(T) were 69.5 mol% and 67.7 mol%, respectively. The major fatty acids of these strains were C16 : 0, C18 : 0, C19 : 0 cyclo ω8c, C18 : 1 2-OH and summed features 3 and 8, consisting of C16 : 1ω7c/C16 : 1ω6c and C18 : 1ω7c/C18 : 1ω6c, respectively. Polar lipid profiles of both of these isolates include diphosphatidylglycerol (DPG), phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylglycerol (PG) and most importantly sulfoquinovosyldiacylglycerol (SQDG), which is one of the typical molecular features reported to delineate the members of the genus Aureimonas from their closest relatives, species of the genus Aurantimonas. Based on the phylogenetic, phenotypic and chemotaxonomic features, strains CC-CFT023(T) and CC-CFT034(T) represent two novel species within the genus Aureimonas, for which the names Aureimonas ferruginea sp. nov. (type strain CC-CFT023(T) = BCRC 80439(T) = JCM 18444(T)) and Aureimonas rubiginis sp. nov. (type strain CC-CFT034(T) = BCRC 80440(T) = JCM 18445(T)) are proposed.

A naphthalene-degrading isolate able to utilize naphthalene as a sole carbon source was identified as Gordonia sp. CC-NAPH129-6. Here a detail characterization of the naphthalene catabolic genes present in this strain was conducted. In nar region four structural genes (narAa, narAb, narB, narC), two regulatory genes (narR1, narR2), a rubredoxin encoding gene (rub1) and a gene (orf7) with unknown function were obtained. When compared with most of the members within naphthalene-degrading Rhodococcus, these naphthalene catabolic genes in strain CC-NAPH129-6 were organized into an operon-like gene cluster and present in the same order. This naphthalene gene cluster located in a 97-kb small plasmid of strain CC-NAPH129-6, as can be seen from the PFGE and Southern blot hybridization data. Besides, a partial transposase sequence containing an IS element structure with 12-nt inverted repeat at both ends was found, which was flanked by direct repeats downstream the narC gene in strain CC-NAPH129-6. This novel transposase gene sequence was unlike to the transposase sequence found between narR2 and rub1 genes in Rhodococcus opacus R7. The comparative analyses of the naphthalene catabolic genes, 16S rRNA and gyrB gene present in strain CC-NAPH129-6 and naphthalene-degrading Rhodococcus species imply that the naphthalene catabolic genes in strain CC-NAPH129-6 might be horizontally transferred from Rhodococcus members. This is the first report demonstrating that naphthalene catabolic genes organized into an operon-like gene cluster in the genus Gordonia, and this might provide evidence of the importance of this actinobacterial lineage in the bioremediation of oil-contaminated soils., (Copyright © 2011 Elsevier GmbH. All rights reserved.)

In the present study, a yellow-pigmented, Gram-negative, short rod-shaped novel bacterium that was capable of degrading a wide range of polycyclic aromatic hydrocarbons (naphthalene, phenanthrene and pyrene) was isolated from agricultural soil located in Yunlin County, Taiwan. Comparative 16S rRNA gene sequence analysis positioned the novel strain in the genus Sphingomonas as an independent lineage adjacent to a subclade containing Sphingomonas fennica K101(T), Sphingomonas histidinilytica UM2(T), Sphingomonas wittichii RW1(T) and Sphingomonas haloaromaticamans A175(T). 16S rRNA gene sequence analysis of strain CC-Nfb-2(T) showed highest sequence similarity to S. fennica K101(T) (96.2%), S. histidinilytica UM2(T) (96.1%), S. wittichii RW1(T) (95.9%), S. haloaromaticamans A175(T) (95.7%), and Sphingobium ummariense RL-3(T) (94.7%); lower sequence similarities were observed with strains of all other Sphingomonas species. The strain contained phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, sphingoglycolipid and diphosphatidylglycerol. The predominant fatty acids were summed feature 8 (C(18:1)ω7c and/or C(18:1)ω6c) C(16:0) and 11-methyl C(18:1)ω7c; C(14:0) 2-OH was the major 2-hydroxy fatty acid. Previously, these lipids have been found to be characteristic of members of the genus Sphingomonas. The serine palmitoyl transferase gene (spt) was also detected and sphingolipid synthesis was confirmed. The predominant isoprenoid quinone system was ubiquinone (Q-10) and the isolate contained sym-homospermidine as the major polyamine. The DNA G+C content of the isolate was 62.8±0.8 mol%. On the basis of chemotaxonomic, phenotypic and phylogenetic data, strain CC-Nfb-2(T) represents a novel species within the genus Sphingomonas, for which the name Sphingomonas formosensis sp. nov. is proposed; the type strain is CC-Nfb-2(T) (=BCRC 80272(T)=DSM 24164(T)).

A gram-negative, spiral or rod-shaped, non-spore-forming diazotrophic bacterium, designated CC-Nfb-7(T), was isolated from agricultural soil in Yunlin County, Taiwan. 16S rRNA gene sequence analysis showed that strain CC-Nfb-7(T) was most closely related to Azospirillum brasilense DSM 1690(T) (97.4 % 16S rRNA gene sequence similarity), Azospirillum rugosum IMMIB AFH-6(T) (96.8 %) and Azospirillum oryzae JCM 21588(T) (96.6 %); <96.5 % 16S rRNA gene sequence similarity was found with all other members of the genus Azospirillum. DNA-DNA relatedness between strain CC-Nfb-7(T) and A. brasilense DSM 1690(T), A. rugosum DSM 19657(T) and A. oryzae JCM 21588(T) was 38.9, 30.1 and 31.8 %, respectively. The respiratory quinone was ubiquinone Q-10. The major fatty acids were summed feature 8 (consisting of C(18 : 1)ω7c and/or C(18 : 1)ω6c), summed feature 3 (consisting of C(16 : 1)ω7c and/or C(16 : 1)ω6c), summed feature 2 (consisting of C(14 : 0) 3-OH and/or iso-C(16 : 1) I), C(16 : 0), C(18 : 0) 2-OH and C(16 : 0) 3-OH. The polar lipids consisted mainly of phosphatidylglycerol, phosphatidylcholine and one unidentified phospholipid. Furthermore, moderate amounts of phosphatidylethanolamine, phosphatidyldimethylethanolamine and one unidentified aminophospholipid were also detected. Strain CC-Nfb-7(T) could be distinguished from members of phylogenetically related species by differences in phenotypic properties. On the basis of morphological, chemotaxonomic and phylogenetic data, strain CC-Nfb-7(T) represents a novel species within the genus Azospirillum, for which we propose the name Azospirillum formosense sp. nov. The type strain is CC-Nfb-7(T) ( = BCRC 80273(T) = JCM 17639(T) = DSM 24137(T)).

Bioremediation, a strategy mediated by microorganisms, is a promising way used in the degradation or removal of organic contaminants from soil or aquatic system. Exopolysaccharide (EPS) which was produced by a variety of Gram-negative bacteria has been demonstrated to be a potential bioemulsifier used in the degradation of hydrocarbons. In the present study, attempts were made to optimize the production of EPS from our newly isolates by adjusting the culture conditions and medium components. Besides, the performance of diesel oil emulsification using partially purified EPS derived from different conditions was also demonstrated. Out of 40 root nodulating bacteria the better emulsifying abilities were recorded from three strains namely Rhizobium miluonense CC-B-L1, Burkholderia seminalis CC-IDD2w and Ensifer adhaerens CC-GSB4, as can be seen from their emulsification index (E(24)) 66, 64 and 60%, respectively. These three strains produced 212, 203 and 198 mg l(-1) of EPS, respectively, in yeast extract mannitol (YEM) medium. After modifying culture conditions, better performances can be achieved from these three strains, with increases of 21.7, 21.4, 16.7% in the EPS production and 12.1, 10.9, 8.3% in E(24), respectively. When considered for strain CC-B-L1 and CC-IDD2w, the addition of 1.5% (v/v) of mannitol and 0.1% (v/v) of asparagine in YEM enhanced 42.9 and 34.7% in EPS production along with 28.8 and 37.5% higher in E(24). The supplement of 2.0% (v/v) glucose and 0.2% (v/v) asparagine in YEM increased 65.2% of EPS and 38.3% of E(24) in strain CC-GSB4. This is the first report demonstrating the optimization of diesel emulsification by EPS from root nodulating isolates, and these microbial agents might be used in the remediation of hydrocarbon contaminated soils in a near future.

The lexA-imuB-dnaE2 gene cassette contributing to the TLS (translesion synthesis) polymerase activity and can easily cause mutation after DNA damage in many bacteria. But it was previously thought that TLS polymerase activity was unlikely to exist in the radio-resistant genus Deinococcus. In our preliminary studies, the lexA-imuB-dnaE2 gene cassette was found in a newly isolated feather-degrading Deinococcus ficus. Here we have attempted to determine the imuB gene sequence from another Deinococcus species namely D. grandis, by using the newly designed primers. The destroying of either imuB or dnaE2 gene in D. ficus leads to the increase in UV sensitivity and decrease in UV-induced mutations, which demonstrated the existence of TLS polymerase activity in D. ficus. In the presence of lexA-imuB-dnaE2, it is possible to obtain mutants with various keratinolytic activities after UV exposure. The keratinolytic activity of mutant strain CC-ZG207 increased by approximately twofold during growth in liquid feather medium. In contrast, the mutant strain CC-ZG227 showed only half of the keratinolytic activity compared with the wild type strain. By utilizing SDS-PAGE and zymogram profile analysis, the change in the protease activity was observed. We have proposed that the superior mutants of D. ficus can be created under UV stress, which is mediated by the lexA-imuB-dnaE2 gene cassette., (Copyright © 2011 Elsevier GmbH. All rights reserved.)

A beige-coloured, gram-staining negative, aerobic, non-motile, moderately thermophilic, rod-shaped bacterium, CC-MHSW-1(T), was isolated on Marine Agar 2216 from a water sample from a coastal hot spring on Green Island (Lutao), located off Taituang, Taiwan. 16S rRNA gene sequence analysis demonstrated that the novel strain shared <94 % sequence similarity with members of the genera Lutimaribacter, Maritimibacter and Oceanicola. Ubiquinone (Q-10) was the major respiratory quinone and C(18 : 1)ω7c was the predominant fatty acid. The predominant polar lipids were phosphatidylcholine, phosphatidylglycerol, phosphatidylethanolamine, diphosphatidylglycerol and unidentified phospholipids and aminolipids. The DNA G+C content of strain CC-MHSW-1(T) was 64.7 mol%. On the basis of 16S rRNA gene sequence analysis and the chemotaxonomic and physiological data, strain CC-MHSW-1(T) represents a new genus and species in the family Rhodobacteraceae for which the name Jhaorihella thermophila gen. nov., sp. nov., is proposed. The type strain of the type species is CC-MHSW-1(T) ( = JCM 15068(T) = CCM 7767(T)).

The modern agricultural practice utilizing plant growth promoting rhizobacteria (PGPR) has brought great benefits in the promotion of crop growth. Among PGPR, Azospirillum is considered as an important genus which is not only closely-associated with plants but also shows potential in the degradation of organic contaminants. However, lack of media for selective isolation or techniques for specific detection or identification limit the exploration of these rhizobacteria. This motivated us to design a genus-specific oligonucleotide primer pair which could assist in rapid detection of species of the genus Azospirillum by means of PCR-specific amplification. The sensitivity and specificity of the newly designed primer pair Azo494-F/Azo756-R were tested against 12 Azospirillum type strains and other closely-related genera. The Azospirillum-specific 16S rRNA gene fragment (263 bp) was successfully amplified for all the reference Azospirillum species with the designed primer pair. No amplification was noted for closely-related species from other genera. The genus specificity was validated with 18 strains including environmental isolates. Interestingly, two strains assigned earlier as Azospirillum amazonense (DSM 2787(T)) and Azospirillum irakense (DSM 11586(T)) failed to produce an Azospirillum-specific fragment with this primer pair. Further phylogenetic analysis of these two isolates based on 16S rRNA gene sequences shows that these two strains might belong to other genera rather than Azospirillum. Preliminary screening of isolates and soil samples with the Azospirillum-specific primers was successful in terms of the rapid detection of Azospirillum isolates. By using real-time PCR analysis the minimum limit of Azospirillum detection was 10(2) CFU g(-1) in the seeded soil sample. The newly designed primers can be used to study the diversity of Azospirillum in ecosystems and aid in the exploration of novel species.

A yellow-pigmented, Gram-negative, rod-shaped, non-spore-forming bacterium, strain CC-TPE-1(T), was isolated from oil-contaminated soil near an oil refinery located in Kaohsiung County, Taiwan. 16S rRNA gene sequence analysis of strain CC-TPE-1(T) showed highest sequence similarity to Novosphingobium naphthalenivorans TUT562(T) (98.1 %), N. panipatense SM16(T) (97.9 %) and N. mathurense SM117(T) (97.6 %) and lower (<97 %) sequence similarity to all other Novosphingobium species. DNA-DNA hybridizations of strain CC-TPE-1(T) with N. naphthalenivorans DSM 18518(T), N. panipatense SM16(T) and N. mathurense SM117(T) showed low relatedness of 30 % (reciprocal 35 %), 29.1 % (reciprocal 30.6 %) and 35 % (reciprocal 23.6 %), respectively. The major respiratory quinone was ubiquinone Q-10, the predominant fatty acid was C(18 : 1)ω7c (49.9 %) and three 2-hydroxy fatty acids, C(14 : 0) 2-OH (8.2 %), C(15 : 0) 2-OH (2.45 %) and C(16 : 0) 2-OH (1.05 %), were detected. Polar lipids consisted mainly of phosphatidylethanolamine, phosphatidylglycerol, phosphatidylcholine, phosphatidyldimethylethanolamine, two sphingoglycolipids, phosphatidylmonomethylethanolamine and several unidentified lipids, and a yellow pigment was also detected. The polyamine pattern contained the single major compound spermidine. Characterization by 16S rRNA gene sequence analysis, physiological parameters, pigment analysis and polyamine, ubiquinone, polar lipid and fatty acid compositions revealed that strain CC-TPE-1(T) represents a novel species of the genus Novosphingobium, for which we propose the name Novosphingobium soli sp. nov., with the type strain CC-TPE-1(T) (=DSM 22821(T) =CCM 7706(T) =CCUG 58493(T)).

A Gram-stain-positive, non-endospore-forming actinobacterium (CC-12301(T)) was isolated from soil attached to a spawn used in the laboratory to grow the edible mushroom Agaricus brasiliensis. Based on 16S rRNA gene sequence similarities, strain CC-12301(T) was shown to belong to the genus Gordonia and was most closely related to the type strains of Gordonia hydrophobica (97.6 % similarity), Gordonia terrae (97.5 %), Gordonia amarae (97.5 %) and Gordonia malaquae (97.4 %). The quinone system was determined to consist predominantly of menaquinone MK-9(H(2)), minor amounts of MK-8(H(2)) and MK-7(H(2)). The polar lipid profile consisted of the major compounds diphosphatidylglycerol and phosphatidylethanolamine, moderate amounts of two phosphatidylinositol mannosides and phosphatidylinositol and minor amounts of phosphatidylglycerol, three unidentified glycolipids, two phosphoglycolipids and a phospholipid. Mycolic acids were present. These chemotaxonomic traits and the major fatty acids, which were C(16 : 1) cis9, C(16 : 0), C(18 : 1) and tuberculostearic acid (10-methyl C(18 : 0)), supported the affiliation of strain CC-12301(T) to the genus Gordonia. The results of physiological and biochemical tests allowed clear phenotypic differentiation of strain CC-12301(T) from the most closely related Gordonia species. Strain CC-12301(T) therefore represents a novel species, for which the name Gordonia humi sp. nov. is proposed, with the type strain CC-12301(T) (=DSM 45298(T) =CCM 7727(T)).

A Gram-negative, beige-pigmented, aerobic, motile, club-shaped bacterium, designated strain CC-SBABM117(T), was isolated from the stipe of the edible mushroom Agaricus blazei Murrill. 16S rRNA gene sequence analysis demonstrated that the strain shared <93 % similarity with the type strains of species in the genera Pannonibacter, Methylopila, Nesiotobacter and Stappia. The organism was unable to produce acid from carbohydrates, but utilized a number of organic acids and amino acids. Ubiquinone 10 (Q-10) was the major respiratory quinone and C(18 : 1) ω 7c, C(19 : 0) cyclo ω 8c, C(16 : 0) and C(18 : 0) were the predominant fatty acids. The predominant polar lipids were diphosphatidylglycerol, phosphatidylcholine, phosphatidylglycerol and phosphatidylethanolamine. The DNA G+C content of strain CC-SBABM117(T) was 62.7 mol%. On the basis of 16S rRNA gene sequence analysis and chemotaxonomic and physiological data, strain CC-SBABM117(T) is considered to represent a novel species of a new genus, for which the name Agaricicola taiwanensis gen. nov., sp. nov. is proposed. The type strain of Agaricicola taiwanensis is CC-SBABM117(T) (=BCRC 17964(T) =CCM 7684(T)).

The taxonomic position of a bright orange-pigmented bacterial strain, designated CC-GZM-130(T), isolated from a water sample of the Guan-zing-ling hot spring, southern Taiwan, was studied. The strain was able to grow on nutrient agar at 25-40 degrees C and in the presence of 1-3 % (w/v) NaCl. Comparative analyses of the 16S rRNA gene sequence showed that the isolate was grouped in the vicinity of the genus Aquiflexum with the highest sequence similarity of 92.1 % to the type strain of Aquiflexum balticum, followed by sequence similarities of 92.0, 91.6 and 91.5 % to the type strains of Algoriphagus ornithinivorans, Algoriphagus hitonicola and Belliella baltica, respectively. The polyamine pattern showed that the major compound was sym-homospermidine. The quinone system was menaquinone MK-7. The polar lipid profile was composed predominantly of phosphatidylethanolamine, three polar lipids and one aminolipid. Minor amounts of other lipids were also detectable. The main characteristics of the fatty acid profiles of strain CC-GZM-130(T), B. baltica and Aquiflexum balticum were similar, with iso-C(15 : 0), iso-C(17 : 1)ω 9c and iso-C(17 : 0) 3-OH as the major fatty acids, but some qualitative and quantitative differences were observed. The DNA G+C content of the novel strain was 53.2 mol%. The isolate clearly differed genotypically and phenotypically from representatives of the most closely related genera. On the basis of these differences, a novel species in a new genus, Fontibacter flavus gen. nov., sp. nov., is proposed with CC-GZM-130(T) (=CCUG 57694(T)=CCM 7650(T)) as the type strain of the type species.

A Gram-stain-positive, coccoid, non-endospore-forming actinobacterium (strain CC-12602(T)) was isolated from a spawn used for growing the edible mushroom Agaricus brasiliensis in the laboratory. On the basis of 16S rRNA gene sequence analysis, strain CC-12602(T) was shown to belong to the genus Microlunatus and was related most closely to the type strains of Microlunatus ginsengisoli (96.1 % similarity), M. phosphovorus (95.9 %), M. panaciterrae (95.8 %) and M. aurantiacus (95.5 %). The quinone system comprised menaquinone MK-9(H4) as the major component and the polyamine pattern consisted of spermidine and spermine as major compounds. The predominant polar lipids were phosphatidylglycerol and unknown phospholipid PL3. Moderate amounts of diphosphatidylglycerol, an unknown glycolipid and three unknown phospholipids and minor amounts of an unknown phospholipid and a polar lipid were detected. The peptidoglycan type was A3gamma', based on LL-2,6-diaminopimelic acid with an interpeptide bridge consisting of a single glycine residue and a second glycine residue at position 1 of the peptide subunit. Peptidoglycan structure and major fatty acids (anteiso-C(15 : 0), iso-C(16 : 0) and iso-C(15 : 0)) supported the affiliation of strain CC-12602(T) to the genus Microlunatus. The results of physiological and biochemical tests allowed strain CC-12602(T) to be differentiated phenotypically from recognized Microlunatus species. Strain CC-12602(T) is therefore considered to represent a novel species of the genus Microlunatus, for which the name Microlunatus soli sp. nov. is proposed. The type strain is CC-12602(T) (=DSM 21800(T) =CCM 7685( T)).

Three Gram-positive, rod-shaped bacteria (strains CC-SBCK-209( T), CC-12309(T) and CC-5209(T)) were isolated from the stalk of the edible mushroom Agaricus blazei grown in the laboratory. 16S rRNA gene sequence analysis indicated that all three isolates clearly belonged to the genus Microbacterium. Strains CC-SBCK-209( T) and CC-12309(T) were most related closely to the type strain of Microbacterium halotolerans (95.9 and 96.1 % 16S rRNA gene sequence similarity, respectively). These two novel strains shared 97.9 % 16S rRNA gene sequence similarity. Levels of similarity to the type strains of all other recognized Microbacterium species were lower than 95.5 %. The third strain (CC-5209( T)) showed the highest 16S rRNA gene sequence similarity to the type strain of Microbacterium resistens (97.6 %); levels of similarity to the type strains of all other recognized Microbacterium species were lower than 96 %. The quinone systems of strains CC-SBCK-209(T), CC-12309(T) and CC-5209(T) consisted of MK-11/MK-12, MK-11/MK-10 and MK-13 as major compounds, respectively. All three strains contained ornithine in their peptidoglycan. The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol and an unknown glycolipid. The polyamine pattern consisted of spermidine and spermine as predominant components. Fatty acid profiles (anteiso-C(15 : 0), iso-C(16 : 0) and anteiso-C(17 : 0 ) as major components) supported the affiliation of all three strains to the genus Microbacterium. The results of physiological and biochemical tests and DNA-DNA hybridization experiments allowed the clear phenotypic and genotypic differentiation of strains CC-SBCK-209(T) and CC-12309( T) from M. halotolerans and other closely related Microbacterium species. Strain CC-5209(T) could be differentiated clearly from M. resistens both genotypically and phenotypically. Based on these data, the novel strains are considered to represent three novel species of the genus Microbacterium. The names proposed for these organisms are Microbacterium agarici sp. nov. [type strain CC-SBCK-209( T) (=DSM 21798(T)=CCM 7686(T))], Microbacterium humi sp. nov. [type strain CC-12309(T) (=DSM 21799(T)=CCM 7687(T))] and Microbacterium pseudoresistens sp. nov. [type strain CC-5209(T) (=DSM 22185(T)=CCM 7688(T))].

The alkB gene encodes for alkane 1-monooxygenase, which is a key enzyme responsible for the initial oxidation of inactivated alkanes. This functional gene can be used as a marker to assess the catabolic potential of bacteria in bioremediation. In the present study, a pair of primers was designed based on the conserved regions of the AlkB amino acid sequences of Actinobacteria, for amplifying the alkB gene from the genus Gordonia (20 Gordonia strains representing 13 species). The amplified alkB genes were then sequenced and analyzed. In the phylogenetic tree based on the translated AlkB amino acid sequences, all the Gordonia segregated clearly from other closely related genera. The sequence identity of the alkB gene in Gordonia ranged from 58.8% to 99.1%, which showed higher sequence variation at the inter-species level compared with other molecular markers, such as the 16S rRNA gene (93.1-99.8%), gyrB gene (77.5-97.3%) or catA gene (72.4-99.5%). The genetic diversity of four selected loci also showed that the alkB gene might have evolved faster than rrn operons, as well as the gyrB or catA genes, in Gordonia. All the available actinobacterial alkB gene sequences derived from the whole genome shotgun sequencing projects are phylogenetically characterized here for the first time, and they exclude the possibility of horizontal gene transfer of the alkB gene in these bacterial groups., (Copyright 2009 Elsevier GmbH. All rights reserved.)

A motile, Gram-negative, pinkish red-pigmented bacterium (strain CC-Hsuan-617(T)) was isolated from an oil-contaminated soil near an oil refinery located in Kaohsiung County, Taiwan. 16S rRNA gene sequence analysis showed that strain CC-Hsuan-617(T) clustered with Algoriphagus mannitolivorans (97.5 % sequence similarity), Algoriphagus aquatilis (98.0 %) and Algoriphagus boritolerans (97.5 %), followed by Algoriphagus ornithinivorans (97.1 %) and Algoriphagus alkaliphilus (97.1 %). The fatty acid profile of the novel strain was slightly different from those reported for recognized Algoriphagus species. The quinone system contained menaquinone MK-7 as the predominant component. The major polar lipids were phosphatidylethanolamine, an unknown polar lipid, an unknown phospholipid and an unknown aminophospholipid. The main cell polyamine was sym-homospermidine; lesser amounts of spermine and spermidine were also found. The results of DNA-DNA hybridization, and physiological and biochemical tests allowed the genotypic and phenotypic differentiation of strain CC-Hsuan-617(T) from recognized Algoriphagus species. Strain CC-Hsuan-617(T) is thus considered to represent a novel species of the genus Algoriphagus, for which the name Algoriphagus olei sp. nov. is proposed. The type strain is CC-Hsuan-617(T) (=BCRC 17886(T)=CCUG 57471(T)).

A yellow-pigmented, Gram-staining-negative, aerobic, non-motile, moderately thermophilic, rod-shaped bacterium, designated strain CC-HSB-11(T), was isolated from a coastal hot spring of Green Island (Lutao), located off Taituang, Taiwan. 16S rRNA gene sequence analysis demonstrated that it shared <94.4 % sequence similarity with Muricauda species. Menaquinone with six isoprene units (MK-6) was the major respiratory quinone and iso-C(15 : 0), iso-C(15 : 1) G, iso-C(15 : 0) 3-OH, iso-C(16 : 0) 3-OH, iso-C(17 : 0) 3-OH and summed feature 3 (comprising iso-C(15 : 0) 2-OH and/or C(16 : 1)omega7c/t) were the predominant fatty acids. The predominant polar lipids were diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine. Six unidentified phospholipids and glycolipids also occurred as minor components. The DNA G+C content of strain CC-HSB-11(T) was 46.4+/-1 mol%. On the basis of 16S rRNA gene sequence similarities with other Muricauda species and differentiating fatty acid compositions and other phenotypic data, strain CC-HSB-11(T) represents a novel species in the genus Muricauda, for which the name Muricauda lutaonensis sp. nov. is proposed. The type strain is CC-HSB-11(T) (=BCRC 17850(T) =KCTC 22339(T)).

Strain CC-SAL-25(T), a non-motile, Gram-negative, red-pigmented bacterium, was isolated from a rare mud-volcano, located in Wandan, Pingtung County, Taiwan. Studies based on 16S rRNA gene sequences showed that the strain clustered closely with Belliella baltica BA143(T) (95.4 % sequence similarity). A menaquinone with seven isoprene units (MK-7) was the major respiratory quinone. The fatty acid profile was slightly different from that of B. baltica BA143(T). The results of the physiological and biochemical tests showed that strain CC-SAL-25(T) could be clearly differentiated from recognized Belliella species based on phenotypic properties. It was evident from the genotypic and phenotypic data that strain CC-SAL-25(T) should be classified as representing a second novel species in the genus Belliella. The name proposed for this taxon is Belliella pelovolcani sp. nov., and the type strain is CC-SAL-25(T) (=BCRC 17883(T)=KCTC 13248(T)).

The C12O gene (catA gene) encodes for catechol 1,2-dioxygenase, which is a key enzyme involved in the first step catalysis of the aromatic ring in the ortho-cleavage pathway. This functional gene can be used as a marker to assess the catabolic potential of bacteria in bioremediation. C12OF and C12OR primers were designed based on the conserved regions of the CatA amino acid sequence of Actinobacteria for amplifying the catA gene from the genus Gordonia (16 Gordonia representing 11 species). The amplified catA genes (382bp) were sequenced and analyzed. In the phylogenetic tree based on the translated catA amino acid sequences, all the Gordonia segregated clearly from other closely related genera. The sequence similarity of the catA gene in Gordonia ranged from 72.4% to 99.5%, indicating that the catA gene might have evolved faster than rrn operons or the gyrB gene at the inter-species level. A single nucleotide deletion of the catA gene was observed in Gordonia amicalis CC-MJ-2a, Gordonia rhizosphera and Gordonia sputi at nucleotide position 349. This deletion led to an encoding frame shift downstream of 11 amino acid residues, from WPSVAARAPAP to GHPWRPAHLHL, which was similar to most of the non-Gordonia Actinobacteria. Such variations might influence the catabolic activities or substrate utilization patterns of catechol 1,2-dioxygenase among Gordonia.

A yellow-pigmented, Gram-staining-negative, aerobic, non-motile, moderately thermophilic, rod-shaped bacterium, CC-MHSW-2T, was isolated from a coastal hot spring of Green Island (Lutao), located off Taituang, Taiwan. The 16S rRNA gene sequence analysis demonstrated that it shared <93.2% sequence similarity with Aquimarina species. The organism was unable to produce acid from carbohydrates, but it could utilize a number of organic acids and amino acids. Menaquinone 6 (MK-6) was the major respiratory quinone and iso-C15:0, iso-C17:0 3-OH and summed feature 3 (comprising C16:1omega7c and/or C16:1omega6c) were the predominant fatty acids. This fatty acid profile distinguished the isolate from members of the genera Aquimarina, Tamlana, Zhouia, Leeuwenhoekiella and Cellulophaga. The DNA G+C content of strain CC-MHSW-2T was 39.7+/-1 mol%. On the basis of the 16S rRNA gene sequence analysis and the chemotaxonomic and physiological data, strain CC-MHSW-2T represents a novel genus and species in the family Flavobacteriaceae, for which the name Lutaonella thermophila gen. nov., sp. nov. is proposed. The type strain is CC-MHSW-2T (=KCTC 22538T=JCM 15069T).

Strain CC-MMS-1(T), a motile, Gram-negative, orange-pigmented bacterium, was isolated from water samples collected from a rare coastal hot spring at Lutao, an island off the eastern coast of Taiwan. 16S rRNA gene sequence studies showed that the novel strain was closely related to Parvularcula bermudensis HTCC2503(T) (95.1 % sequence similarity). The fatty acid profile was slightly different from P. bermudensis HTCC2503(T). The results of physiological and biochemical tests allowed clear phenotypic differentiation of the novel isolate from P. bermudensis HTCC2503(T). It is evident from the genotypic and phenotypic data that the strain should be classified as a novel species in the genus Parvularcula. The name proposed for this taxon is Parvularcula lutaonensis sp. nov. The type strain is CC-MMS-1(T) (=BCRC 17814(T)=KCTC 22245(T)).

A polyphasic taxonomic study was performed on a pink-coloured unknown bacterium isolated from discarded road tar. Comparative analysis of the 16S rRNA gene sequence demonstrated that the isolate belongs phylogenetically to the genus Azospirillum with Azospirillum lipoferum, A. melinis and A. rugosum as its closest phylogenetic relatives (96.7, 96.6 and 96.6 % similarity to the respective type strains). The generic assignment was confirmed on the basis of chemotaxonomic data, which revealed a fatty acid profile characteristic for the genus Azospirillum, consisting of straight-chain saturated and unsaturated fatty acids, with C(18 : 1)omega7c as the major unsaturated non-hydroxylated fatty acid, and C(16 : 0) 3-OH as the major hydroxylated fatty acid, and a ubiquinone with ten isoprene units (Q-10) as the predominant respiratory quinone. On the basis of both the phenotypic and molecular genetic evidence, it is proposed that the unknown isolate should be classified within a novel species of the genus Azospirillum, for which the name Azospirillum picis sp. nov. is proposed. The type strain is IMMIB TAR-3(T) (=CCUG 55431(T) =DSM 19922(T)).

This study was undertaken to assess genetic diversity among Gordonia species present in heavily oil-contaminated sites using both a culture-dependent and a culture-independent (PCR-denaturing gradient gel electrophoresis (DGGE)) approach. Soil samples for this purpose were collected from 8 different heavily (crude) oil-contaminated industrial park sites located around Kaohsiung County, Taiwan. Using Gordonia-specific PCR-DGGE, a significant increase in Gordonia species diversity was noted in 1% heavily oil-enriched soil. A total of 67 strains were scored and identified as Gordonia after genus-specific PCR amplification and sequencing. BOX-PCR fingerprinting of culturable Gordonia showed wide strain diversity. A total of 33 different strains were identified from most of the sampling sites. Based on gyrB gene sequence analysis, all Gordonia strains could be segregated into five major clusters. Gordonia amicalis was the predominant species in all oil-amended soil samples. Isolates sharing <98.5% gyrB gene sequence similarities with Gordonia type strains represent indigenous novel Gordonia species. Variations in phenotypic characteristics further confirm the presence of a wide range of species and strain diversity among Gordonia isolates. Based on the genotypic and phenotypic details obtained here, we conclude that heavily oil-contaminated soil supports diverse indigenous Gordonia strains.

The taxonomic status of a pale-yellow-coloured bacterial isolate from rhizosphere soil of Fortunella hindsii (Champ. ex Benth.) Swingle was characterized using a polyphasic taxonomic approach. Comparative analysis of the 16S rRNA gene sequence showed that the isolate constituted a distinct branch within the genus Sphingobium. The generic assignment was confirmed by chemotaxonomic data, which revealed the presence of a fatty acid profile that was characteristic for the genus Sphingobium, consisting of straight-chain saturated and unsaturated as well as 2-OH fatty acids and the lack of 3-OH fatty acids, ubiquinone with ten isoprene units (Q-10) as the predominant respiratory quinone, and a polar lipid pattern that consisted of the predominant compounds phosphatidylethanolamine, phosphatidylglycerol, phosphatidylmonomethylethanolamine, phosphatidyldimethylethanolamine, diphosphatidylglycerol, sphingoglycolipid and an unknown glycolipid. Spermidine was the major polyamine component. The genotypic and phenotypic data (physiology and fatty acid and polar lipid profiles) showed that the isolate merits classification as representing a novel species of the genus Sphingobium, for which the name Sphingobium rhizovicinum sp. nov. is proposed. The type strain is CC-FH12-1T (=CCM 7491(T)=BCRC 17770T) [corrected]

A Gram-type negative or variable, aerobic, rod-shaped, nitrogen-fixing bacterium, designated strain CC4(T), was isolated on nutrient agar from a soil sample collected at a regional agricultural research field station located in Kaohsiung County, Taiwan. 16S rRNA gene sequence analysis demonstrated that this isolate is unique, showing <92.5 % similarity with respect to species of the genera Xanthobacter (maximum of 92.2 % similarity), Prosthecomicrobium (92.0 %), Devosia (91.9 %), Kaistia (91.9 %) and Methylocystis (91.9 %). The organism utilized acetic acid, formic acid, beta-hydroxybutyric acid, dl-lactic acid, succinic acid, bromosuccinic acid, l-alaninamide, l-alanine, l-alanyl glycine, l-glutamic acid and l-proline as substrates, but not methanol or methylamine. Chemotaxonomic data revealed that strain CC4(T) contains ubiquinone Q-10 as the major respiratory quinone. The characteristic diamino acid of the peptidoglycan was meso-diaminopimelic acid. The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine, phosphatidylmonomethylethanolamine, phosphatidyldimethylethanolamine, an unknown lipid and an unknown aminolipid. The fatty acid profile differed from those of members of the genera Xanthobacter, Prosthecomicrobium, Devosia and Kaistia. The predominant fatty acids were C(18 : 1)omega7c, C(19 : 0) cyclo and C(16 : 0). On the basis of 16S rRNA gene sequence analysis in combination with chemotaxonomic and physiological data, strain CC4(T) represents a novel genus and species, for which we propose the name Pseudoxanthobacter soli gen. nov., sp. nov. The type strain of Pseudoxanthobacter soli is CC4(T) (=DSM 19599(T) =CIP 109513(T)).

Strain CC-SEYA-1T, a motile, Gram-negative, non-violet-pigmented bacterium, was isolated on nutrient agar from spring-water samples collected from Yang-Ming Mountain, Taipei County, Taiwan. 16S rRNA gene sequence studies showed that the strain clustered with Chromobacterium violaceum (96.8 % similarity) and Chromobacterium subtsugae (96.5 % similarity), followed by Aquitalea magnusonii (95.8 % similarity). The fatty acid profile was slightly different from those reported for C. violaceum, C. subtsugae and A. magnusonii. The results of DNA-DNA hybridization, and physiological and biochemical tests allowed both genotypic and phenotypic differentiation of the isolate from the described Chromobacterium species. It is evident from the data obtained that the strain should be classified as a novel species in the genus Chromobacterium. The name proposed for this taxon is Chromobacterium aquaticum sp. nov.; the type strain is CC-SEYA-1T (=CCUG 55175T=BCRC 17769T).

The taxonomic status of a light-orange-coloured bacterial isolate from an oil-contaminated soil sample was characterized by using a polyphasic taxonomic approach. Comparative analysis of 16S rRNA gene sequences demonstrated that the isolate belonged phylogenetically to the genus Azospirillum, with Azospirillum canadense, Azospirillum brasilense and Azospirillum doebereinerae as its closest phylogenetic relatives (97.3, 97.0 and 97.0 % similarity, respectively). DNA-DNA pairing studies showed that the unidentified organism displayed 25.0, 17.0 and 19.0 % relatedness to the type strains of A. brasilense, A. canadense and A. doebereinerae, respectively. The generic assignment was confirmed by chemotaxonomic data, which revealed a fatty acid profile that was characteristic of the genus Azospirillum, consisting of straight-chain saturated and unsaturated fatty acids with C18 : 1 omega 7c as the major fatty acid, and ubiquinone with ten isoprene units (Q-10) as the predominant respiratory quinone. On the basis of both the phenotypic and molecular genetic evidence, it is proposed that the unknown isolate be classified as a representative of a novel species of the genus Azospirillum, for which the name Azospirillum rugosum sp. nov. is proposed. The type strain is IMMIB AFH-6T (=CCUG 53966T=DSM 19657T).

A Gram-negative, short rod-shaped, nitrogen-fixing bacterium (CC-21(T)) was isolated from a soil sample collected from the regional agricultural research station in Kaohsiung County (Taiwan). Using 16S rRNA gene sequence analysis, it could be clearly demonstrated that this isolate was novel: it showed <97 % similarity to species of the genera Acidovorax, Alicycliphilus, Giesbergeria, Simplicispira and Diaphorobacter. The organism used several organic acids, but only a few sugars as substrates. The fatty acid profile differed from those reported for members of the genera Acidovorax, Alicycliphilus, Giesbergeria, Simplicispira and Diaphorobacter. On the basis of 16S rRNA gene sequence analysis in combination with physiological data, strain CC-21(T) represents a novel species in a new genus, for which the name Pseudacidovorax intermedius gen. nov., sp. nov. is proposed; the type strain is CC-21(T) (=CCUG 54492(T)=CIP 109510(T)).