Next-Generation Sequencing in Lung Cancers-A Single-Center Experience in Taiwan.

Background and Objectives: Lung cancer is a leading cause of cancer mortality in Taiwan. With rapid advancement of targeted therapeutics in non-small cell lung cancers, next-generation sequencing (NGS) is becoming an important tool for biomarker testing. In this study, we describe institutional experience of NGS analysis in non-small cell carcinoma (NSCLC). Materials and Methods: A cohort of 73 cases was identified from the institutional pathology archive in the period between November 2020 and December 2022. Results: Adenocarcinoma was the most common histologic type (91.8%). Most patients presented with stage IIIB and beyond (87.7%). Twenty-nine patients (39.7%) were evaluated at the time of initial diagnosis, while the others had received prior chemotherapy or targeted therapy. The most frequently mutated gene was EGFR (63%), and this was followed by TP53 (50.7%), KRAS (13.7%), RB1 (13.7%), and CDKN2A (13.7%). Clinically actionable mutations associated with a guideline-suggested targeted therapy were identified in 55 cases (75.3%) overall, and in 47.1% of cases excluding EGFR TKI-sensitizing mutation. Biomarkers other than EGFR TKI-sensitizing mutations were compared. Cases without TKI-sensitizing EGFR mutation had more level 1 or 2 biomarkers (excluding EGFR TKI-sensitizing mutations) than cases with TKI-sensitizing EGFR mutations (47.1% versus 20.1%, p = 0.016). Progressive disease was associated with co-occurrence of clinically actionable mutations (20.5% versus 0%, p < 0.05). Eight of the nine cases with co-occurring actionable genetic alternations had an EGFR mutation. After an NGS test, 46.1% of actionable or potentially actionable genetic alternations led to patients receiving a matched therapy. Conclusions: Our study demonstrated that NGS analysis identifies therapeutic targets and may guide treatment strategies in NSCLC. NGS tests may be advantageous over multiple single-gene tests for optimization of treatment plans, especially for those with non-EGFR mutations or those with progressive disease.

Effect of seed biopriming with selected endophytes on the growth and chilling tolerance of rice plants.

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.

Description of Gemmobacter aestuarii sp. nov., isolated from estuarine surface water and reclassification of Cereibacter changlensis as Gemmobacter changlensis Chen et al. 2013.

A Gram-stain-negative, tyrosine-metabolizing, non-motile, strictly aerobic, non-spore-forming, rod-shaped marine bacterium, designated strain CC-PW-75T, was isolated from the estuarine water off Pintung, Taiwan. Strain CC-PW-75T formed a distinct phyletic lineage associated with Gemmobacter species, sharing the highest 16S rRNA gene sequence similarity with G. megaterium CF17T and G. straminiformis CAM-8T (96.0% each) followed by G. aquatilis IFAM 1031T and G. nectariphilus AST4T (95.8% each). Analysis of the draft genome (3.76 Mbp) revealed the presence of genes encoding light-harvesting complexes, photosynthetic reaction centers and proteins involved in the metabolism of CO, CO2, HCO3‒ and H2S. However, bacteriochlorophyll a was not detected. Average nucleotide identity values between the genome sequence of CC-PW-75T and the related Gemmobacter species (n = 6) were estimated to be 72.8-76.3%. Polar lipid analysis revealed the presence of phosphatidylglycerol, phosphatidylethanolamine and an unidentified lipid in major amounts, and phosphatidylmonomethylethanolamine, phosphatidylcholine, an unidentified aminolipid and an unidentified lipid in minor amounts. C18:1ω7c and/or C18:1ω6c, C18:0 and C18:1ω7c 11-methyl were identified to be major fatty acids. The DNA G + C content was 66.2 mol% (draft genome sequence). Ubiquinone-10 (Q-10) was the sole respiratory quinone. Based on the polyphasic taxonomic evidence, CC-PW-75T is most likely a novel species of the genus Gemmobacter, affiliated to the family Rhodobacteraceae, for which the name Gemmobacter aestuarii sp. nov. is proposed. The type strain is CC-PW-75T (= JCM 19754T = BCRC 80759T). Also, we propose the reclassification of Cereibacter changlensis as Gemmobacter changlensis Chen et al. 2013 using the polyphasic data presented in this study.

Castellaniella fermenti sp. nov., isolated from a fermented meal.

A polyphasic taxonomic approach was used to characterize a presumably novel bacterium, designated strain CC-YTH191T, isolated from a fermented meal in Taiwan. Cells of strain CC-YTH191T were Gram-stain-negative aerobic rods, which grew at 15-40 °C (optimal 25-30 °C), pH 6.0-9.0 (optimal 7.0) and 1-2 % (w/v) NaCl (optimal 1 %). On the basis of 16S rRNA gene sequence analysis, strain CC-YTH191T appeared to belong to the genus Castellaniella, and was closely related to Castellaniella hirudinis (96.7 % similarity), Castellaniella ginsengisoli (96.7 %) and Castellaniella caeni (96.0 %), while with other related species it shared <96.0 % similarity. The major cellular fatty acids of the isolate were C16 : 0, C17 : 0cyclo, C14 : 0 3OH/C16 : 1iso I and C18 : 1ω7c/C18 : 1ω6c. The polar lipid profile contained diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylserine, three unidentified phospholipids, an unidentified aminolipid and an unidentified aminophospholpid. Putrescine was the predominant polyamine followed by spermidine. The DNA G+C content was 62.2 mol% and the predominant quinone system was ubiquinone 8 (Q-8). All these features confirmed the placement of the strain CC-YTH191T as a novel species within the genus Castellaniella, for which the name Castellaniella fermenti sp. nov. is proposed. The type strain is CC-YTH191T (=BCRC 81023T=JCM 31755T).

Sphingomonas colocasiae sp. nov., isolated from taro (Colocasia esculanta).

A polyphasic approach was used to characterize an aerobic, Gram-stain-negative, rod-shaped bacterium (designed as strain CC-MHH0539T) isolated from the chopped tuber of taro (Colocasia esculanta) in Taiwan. Strain CC-MHH0539T was able to grow at 15-30 °C (optimum, 25 °C), at pH 6.0-9.0 (optimum, 7.0) and with 0-1 % (w/v) NaCl. Strain CC-MHH0539T showed highest 16S rRNA gene sequence similarity to Sphingomonas laterariae LNB2T (96.8 %), Sphingobium boeckii 469T (96.5 %), Sphingomonas faucium E62-3T (96.4 %) and Sphingosinicella vermicomposti YC7378T (96.2 %) and <96.1 % similarity to other sphingomonads. Strain CC-MHH0539T was found to cluster mainly with the clade that accommodated members of the genus Sphingomonas. The dominant cellular fatty acids were C16 : 0, C16 : 1ω5c, C14 : 0 2-OH, C16 : 1ω7c/C16 : 1ω6c and C18 : 1ω7c/C18 : 1ω6c. Diphosphatidylglycerol, phosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine, phosphatidylmonomethylethanolamine, two sphingoglycolipids and two unidentified phospholipids were detected in strain CC-MHH0539T. The DNA G+C content was 69.5 mol%. The respiratory quinone system and predominant polyamine was ubiquinone 10 (Q-10) and sym-homospermidine, respectively, which is in line with Sphingomonas representatives. Based on the distinct phylogenetic, phenotypic and chemotaxonomic traits, strain CC-MHH0539T is considered to represent a novel species of the genus Sphingomonas, for which the name Sphingomonas colocasiae sp. nov. is proposed. The type strain is CC-MHH0539T (=BCRC 80933T=JCM 31229T).

Filimonas aquilariae sp. nov., isolated from agarwood chips.

A polyphasic approach was used to characterize a Gram-staining negative bacterium (designated strain CC-YHH650T) isolated from agarwood chips. Strain CC-YHH650T was aerobic and rod-shaped, able to grow at 15-37 °C (optimal 30 °Ð¡), pH 6.0-8.0 (optimal 7.0) and 0-1 % (w/v) NaCl (optimal 0 %). Phylogenetic analysis based on 16S rRNA genes revealed that strain CC-YHH650T shared highest sequence similarities with Filimonas lacunae (97.5 %), F. zeae (97.4 %), F. endophytica (97.3 %) and F. aurantiibacter (93.0 %), and lower sequence similarity with other genera (less than 93.0 %). The levels of DNA-DNA relatedness between strain CC-YTH209T, F. lacunae, F. endophytica and F. zeae were estimated to be 18.3, 6.1, 24.7 % (the reciprocal values were 9.8, 8.8, 18.3 %). The major fatty acids were iso-C15 : 0, iso-C15 : 1 G, C16 : 0 3-OH, iso-C17 : 0 3-OH and C16 : 1ω7c/C16 : 1ω6c. The polar lipid profile contained phosphatidylethanolamine, an unidentified phospholipid, two unidentified aminophospholipids, three unidentified aminolipids and three unidentified lipids. The DNA G+C content was 46.6 mol% and the predominant quinone was menaquinone-7 (MK-7). The major polyamine was sym-homospermidine. Based on the distinct phylogenetic, phenotypic and chemotaxonomic traits together with results of comparative 16S rRNA gene sequence analysis, strain CC-YHH650T is considered to represent a novel species of the genus Filimonas, for which the name Filimonas aquilariae sp. nov. is proposed. The type strain is CC-YHH650T (=BCRC 80935T=JCM 31197T).

Hydrogenophaga aquatica sp. nov., isolated from a hot spring.

A polyphasic approach was used to characterize an aerobic, Gram-negative, rod-shaped bacterium (designated strain CC-KL-3T) isolated from a hot spring. Phylogenetic analyses based on 16S rRNA genes indicated that strain CC-KL-3T showed highest sequence similarity to Hydrogenophaga bisanensis (97.7 %) and Hydrogenophaga atypica (97.6 %) and lower sequence similarity to other species (less than 97.6 %). The levels of DNA-DNA relatedness between strain CC-KL-3T, H. bisanensis and H. atypica were estimated to be 13.0 and 8.7 % (the reciprocal value was 14.7 and 6.3 %). Strain CC-KL-3T was non-motile, without apparent flagella and able to grow between 15-42 °C (optimal 30 °Ð¡), pH 6.0-8.0 (optimal 7.0) and 0-2 % (w/v) NaCl (optimal 0 %). The DNA G+C content was 61.4 mol% and the major quinone system was ubiquinone (Q-8). The polyamine profile revealed the predominance of 2-hydroxyputrescine and putrescine and the dominant cellular fatty acids were C16 : 0 (28.9 %), C16 : 1ω7c/C16 : 1ω6c (41.4 %) and C18 : 1ω7c/C18 : 1ω6c (11.9 %). These data corroborated the affiliation of strain CC-KL-3T to the genus Hydrogenophaga. Based on the distinct phylogenetic, phenotypic and chemotaxonomic traits, and the results of comparative 16S rRNA gene sequence analysis, strain CC-KL-3T is considered to represent a novel species of the genus Hydrogenophaga, affiliated to the family Comamonadaceae, for which the name Hydrogenophaga aquatica sp. nov. is proposed. The type strain is CC-KL-3T (=BCRC 80937T=JCM 31216T).

Olivibacter composti sp. nov., isolated from compost collected at a greenhouse.

A polyphasic approach was used to characterize a presumably novel bacterium, designated strain CC-KYC063T, isolated from a compost sample collected at a greenhouse facility within Taiwan. Strain CC-KYC063T was Gram-stain-negative and aerobic, able to grow at 15-37 °C (optimal 30 °C), at pH 7.0-9.0 (optimal pH 7.0) and with 0-2 % (w/v) NaCl. The 16S rRNA gene sequence of strain CC-KYC063T showed highest similarity to Olivibacter ginsengisoli Gsoil 060T (96.2 %) and Olivibacter terrae Jip13T (95.5 %). The DNA G+C content was 43.0 mol% and the predominant quinone system was menaquinone-7 (MK-7). The polyamine pattern showed a predominance of sym-homospermidine. The major fatty acids found in strain CC-KYC063T were iso-C15 : 0, iso-C17 : 0 3-OH and C16 : 1ω6c/C16 : 1ω7c. Based on the distinct phylogenetic, phenotypic and chemotaxonomic traits together with results of comparative 16S rRNA gene sequence analysis, strain CC-KYC063T is considered to represent a novel species of the genus Olivibacter, for which the name Olivibacter composti sp. nov. is proposed. The type strain is CC-KYC063T (=BCRC 80939T=JCM 31198T).

Humibacter aquilariae sp. nov., an actinobacterium isolated from an agarwood chip.

A polyphasic taxonomic approach was used to characterize a presumably novel bacterium, designated strain CC-YTH161T, isolated from an agarwood sample. Cells of strain CC-YTH161T were Gram-stain-positive aerobic rods, which grew at 20-40 °C, at pH 5.0-9.0 and with 0-5 % (w/v) NaCl. On the basis of 16S rRNA gene sequence analysis, strain CC-YTH161T appeared to belong to the genus Humibacter, and was closely related to Humibacter antri D7-27T (96.6 % similarity) and Humibacter ginsengiterrae DCY60T (96.2 %). The DNA G+C content was 67.0 mol% and the predominant quinone system was menaquinones (MK) 11 and 12. The major cellular fatty acids of the isolate were C15 : 0, iso-C16 : 0, anteiso-C17 : 0 and C18 : 1ω7c/C18 : 1ω6c. The polar lipid profile comprised predominant amounts of diphosphatidylglycerol followed by two unidentified co-migrating glycolipids and phosphatidylglycerol in significant amounts. The diagnostic diamino acid was 2,4-diaminobutyric acid. All these features confirmed the placement of strain CC-YTH161T within the genus Humibacter. On the basis of evidence from this study, strain CC-YTH161T is considered to represent a novel species of the genus Humibacter, for which the name Humibacter aquilariae sp. nov. is proposed. The type strain is CC-YTH161T (=BCRC 80936T=JCM 31199T).

Ruficoccus amylovorans gen. nov., sp. nov., an amylolytic and nitrate-reducing diazotroph of the family Puniceicoccaceae.

A novel amylolytic, nitrate-reducing and diazotrophic bacterium, designated strain CC-MHH0563T, isolated from a fermenter was assessed for its taxonomic status using a polyphasic approach. Cells of strain CC-MHH0563T were Gram-staining-negative, catalase- and oxidase-positive, mesophilic and aerobic cocci, which produced reddish nondiffusible pigments. Growth was observed at 15-37 °C (optimal 25 °C), pH 6.0-8.0 (optimal pH 7.0) and salinity of 0-3 % (w/v). Strain CC-MHH0563T showed highest pairwise 16S rRNA gene sequence similarity to members of the genera Cerasicoccus(89.3-89.5 %), Coraliomargarita (87.8 %), Pelagicoccus(85.8-86.4 %) and Puniceicoccus (87.9 %), and established a discrete taxonomic lineage during phylogenetic analysis. The major fatty acids found in strain CC-MHH0563T were C14 : 0, anteiso-C15 : 0, C16 : 0, C17 : 0, C18 : 0 and C18 : 1ω9c. The polar lipid profile consisted of phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylglycerol, two unknown aminolipids and two unknown aminophospholipids. The polyamine pattern showed a predominance of spermidine and a minor amount of cadaverine. The DNA G+C content was 57.4 mol% and the predominant quinone system was menaquinone-7. The low 16S rRNA gene sequence similarity values (<90.0 %) and a distinct phylogenetic clustering clearly distinguished strain CC-MHH0563T from other representatives of the family Puniceicoccaceae. Based on the discrete phylogenetic, phenotypic and chemotaxonomic traits together with the results of comparative 16S rRNA gene sequence analysis, strain CC-MHH0563T is considered to represent a novel genus and species of the family Puniceicoccaceae, for which the name Ruficoccus amylovorans gen. nov., sp. nov. is proposed. The type strain of the type species is CC-MHH0563T (=BCRC 80918T=JCM 31066T).

Chryseobacterium endophyticum sp. nov., isolated from a maize leaf.

A Gram-stain-negative, aerobic, yellow-pigmented, flexirubin-producing, rod-shaped and endophytic bacterium, designated strain CC-YTH209T, was isolated from a maize leaf and subjected to a taxonomic study. Strain CC-YTH209T was found to grow at 15-40 °C (optimal 30 °C), at pH 6.0-8.0 (optimal pH 7.0) and in the presence of 0-2 % (optimal 1 %) (w/v) NaCl. On the basis of 16S rRNA gene sequence analysis, strain CC-YTH209T appeared to belong to the genus Chryseobacterium within the class Flavobacteriia, and was closely related to Chryseobacterium rigui CJ16T (97.5 % similarity) and Chryseobacterium taeanense PHA3-4T (96.9 %). The level of DNA-DNA relatedness between strain CC-YTH209T and Chryseobacterium rigui CJ16T was 14.4 % (reciprocal, 13.0 %). Phylogenetic analyses based on 16S rRNA genes revealed a distinct taxonomic position attained by strain CC-YTH209T within the clade that accommodated Chryseobacterium species. The DNA G+C content was 37.2 mol%. Strain CC-YTH209T contained menaquinone MK-6 as the predominant respiratory quinone and sym-homospermidine as the major polyamine. The major cellular fatty acids of the isolate were iso-C15 : 0, iso-C17 : 0 3-OH and C16 : 1ω6c/C16 : 1ω7c. The polar lipid profile comprised phosphatidylethanolamine and five aminolipids, three lipids, one glycolipid, one aminophospholipid and one phospholipid, which are presently uncharacterized. On the basis of evidence presented in this study, strain CC-YTH209T is considered to represent a novel species of the genus Chryseobacterium, for which the name Chryseobacterium endophyticum sp. nov. is proposed. The type strain is CC-YTH209T (=BCRC 80938T=JCM 31226T).

Sphingobacterium cibi sp. nov., isolated from the food-waste compost and emended descriptions of Sphingobacterium spiritivorum (Holmes et al. 1982) Yabuuchi et al. 1983 and Sphingobacteriumthermophilum Yabe et al. 2013.

A novel, pale yellow-coloured, Gram-staining-negative, aerobic, rod-shaped and non-motile bacterium, designated as strain CC-YY411T, was isolated from the food-waste compost and subjected to polyphasic taxonomy. Strain CC-YY411T exhibited the highest pairwise 16S rRNA gene sequence similarity to Sphingobacterium thermophilum JCM 17858T (94.6 %) while sharing 94.1 to 89.7 % similarities with other Sphingobacterium species. Novel strain established a discrete phylogenetic lineage within the clade that accommodated validly established members of the genus Sphingobacterium. The polar lipid profile of strain CC-YY411T contained major amounts of phosphatidylethanolamine, one unidentified lipid and two unidentified aminolipids besides accommodating trace amounts of a sphingolipid, two phospholipids, an unidentified aminophospholipid and an unidentified glycolipid. The DNA G+C content of strain CC-YY411T was 34.5 mol%. The major and minor respiratory quinones were MK-7 (89.1 %) and MK-6 (10.9 %), respectively. The predominant fatty acids were iso-C15 : 0 (24.0 %), iso-C17 : 0 3-OH (11.9 %), iso-C15 : 0 3-OH (6.9 %), C15 : 1ω5c (5.5 %) and summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c) (32.0 %). Based on the phylogenetic, phenotypic and chemotaxonomic distinctiveness, strain CC-YY411T represents a novel species of Sphingobacterium, for which the name Sphingobacteriumcibi sp. nov. is proposed. The type strain is CC-YY411T (=BCRC 80430T=JCM 18407T). Amended species descriptions of Sphingobacterium spiritivorum(Holmes et al. 1982) Yabuuchi et al. 1983 and Sphingobacterium thermophilum Yabe et al. 2013 are also proposed based on new data obtained in this study.

Azospirillum agricola sp. nov., a nitrogen-fixing species isolated from cultivated soil.

A polyphasic approach was used to characterize a novel nitrogen-fixing bacterial strain, designated CC-HIH038T, isolated from cultivated soil in Taiwan. Cells of strain CC-HIH038T were Gram-stain-negative, facultatively aerobic and spiral-shaped, with motility provided by a single polar flagellum. The 16S rRNA gene sequence analysis of strain CC-HIH038T showed highest sequence similarity to Azospirillum doebereinerae (98.0 %), Azospirillum thiophilum (97.5 %), Azospirillum rugosum (97.4 %) and Azospirillum zeae (97.2 %) and lower sequence similarity ( < 97.0 %) to all other species of the genus Azospirillum. According to DNA-DNA association, the relatedness values of strain CC-HIH038T with A. doebereinerae, A. thiophilum, A. rugosum and A. zeae were 51.8 %, 41.2 %, 56.5 % and 37.5 %, respectively. Strain CC-HIH038T was able to grow at 20-37 °C and pH 7.0-8.0. Strain CC-HIH038T gave positive amplification for dinitrogen reductase (nifH gene); the activity was recorded as 8.4 nmol ethylene h- 1. The predominant quinone system was ubiquinone Q-10 and the DNA G+C content was 68.8 mol%. The major fatty acids found in strain CC-HIH038T were C16 : 0, iso-C18 : 0, C16 : 0 3-OH, C14 : 0 3-OH/iso-C16 : 1 and C18 : 1ω7c/C18 : 1ω6c. Based on the distinct phylogenetic, phenotypic and chemotaxonomic traits together with results of comparative 16S rRNA gene sequence analysis, strain CC-HIH038T is considered to represent a novel species in the genus Azospirillum, for which the name Azospirillum agricola sp. nov. is proposed. The type strain is CC-HIH038T ( = BCRC 80909T = JCM 30827T).

Idiomarina tyrosinivorans sp. nov., isolated from estuarine surface water.

A tyrosine-metabolizing, Gram-stain-negative, strictly aerobic, non-spore-forming, curved-rod-shaped, motile (due to monopolar flagellum) marine bacterium, designated strain CC-PW-9T, was isolated from estuarine water off Pingtung, Taiwan. Strain CC-PW-9T not only shared highest 16S rRNA gene sequence similarities with Idiomarina representatives (96.4-93.4 %, n=26), but also formed a distinct phyletic lineage and coherent phylogenetic cluster associated with those species. Cells of strain CC-PW-9T grew with 6-12 % (w/v) NaCl, at 20-40 °C and at pH 6-9. It produced predominant amounts of phosphatidylethanolamine, plus diphosphatidylglycerol, phosphatidylglycerol, phosphatidylserine, two unidentified phospholipids, an unidentified phosphoglycolipid, two unidentified lipids and an unidentified aminolipid in moderate to trace amounts. Fatty acids such as iso-C15 : 0, iso-C17 : 0 and iso-C17 : 1ω9c and/or C16 : 010-methyl (summed feature 9) were found in major amounts. The DNA G+C content was 51.1 mol%. Ubiquinone-8 (Q-8) was the sole respiratory quinone. Based on evidence from this polyphasic taxonomic study, strain CC-PW-9T is considered to represent a novel species of the genus Idiomarina, affiliated to the family Idiomarinaceae, for which the name Idiomarina tyrosinivorans sp. nov. is proposed. The type strain is CC-PW-9T (=JCM 19757T=BCRC 80745T).

Leucobacter zeae sp. nov., isolated from the rhizosphere of maize (Zea mays L.).

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 DNA­DNA 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).

Azospirillum soli sp. nov., a nitrogen-fixing species isolated from agricultural soil.

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).

Bacillus formosensis sp. nov., isolated from pesticide wastewater.

A Gram-stain-positive, endospore-forming rod (designated strain CC-LY275T) was isolated from a pesticide wastewater sample. The isolate grew at a temperature 20-45 °C, at pH 7.0-8.0 and tolerated NaCl 6 % (w/v). The most closely related strains in terms of 16S rRNA gene sequence similarity were Bacillus horneckiae (97.1 %) and Bacillus oceanisediminis (96.8 %), respectively. The G+C content of the genomic DNA was 37.9 mol%. Strain CC-LY275T was determined to possess iso-C14 : 0, iso-C15 : 0 and anteiso-C15 : 0 as predominant fatty acids. The major polar lipid profile consisted of diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylglycerol. Cell-wall peptidoglycan contained meso-diaminopimelic acid; menaquinone (MK-7) was the predominant respiratory quinone. According to the distinct phylogenetic, phenotypic and chemotaxonomic properties, the name Bacillus formosensis sp. nov. (type strain CC-LY275T = BCRC 80443T = JCM 18448T) is proposed.

Paenibacillus medicaginis sp. nov. a chitinolytic endophyte isolated from a root nodule of alfalfa (Medicago sativa L.).

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.