Pseudomonas lopnurensis sp. nov., an endophytic bacterium isolated from Populus euphratica at the ancient Ugan river.

A Gram-stain negative, aerobic, rod-shaped, motile by a single polar flagellum, non-spore-forming bacterium, designated strain AL-54T, was isolated from the storage liquid in the stems of Populus euphratica tree at the ancient Ugan River in Xinjiang, PR China. Isolated AL-54T grew optimally at pH 7.0 and temperature 35 °C in the presence of 3% (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequence demonstrated that the isolate belonged to the genus Pseudomonas and was closely related to Pseudomonas songnenensis NEAU-ST5-5 T (97.6%), Pseudomonas zhaodongensis NEAU-ST5-21 T (97.5%), Pseudomonas alcaliphila AL15-21T (97.3%), Pseudomonas toyotomiensis HT-3T (97.3%), Pseudomonas oleovorans subsp. lubricantis RS1T (97.3%), Pseudomonas stutzeri ATCC 17588T (97.3%), Pseudomonas chengduensis CGMCC 2318T (97.2%), and Pseudomonas xanthomarina KMM 1447T (97.1%). Multilocus Sequences Analysis (MLSA) of strain AL-54T based on the three housekeeping genes, rpoB, rpoD and gyrB further confirmed the phylogenetic assignment of the isolates. The G+C content was 64.7 mol%. The DNA-DNA hybridization with P. songnenensis NEAU-ST5-5 T, P. zhaodongensis NEAU-ST5-21T, P. alcaliphila AL15-21T, P. toyotomiensis HT-3T, P. oleovorans subsp. lubricantis RS1T, P. stutzeri ATCC 17588T, P. chengduensis CGMCC 2318T and P. xanthomarina KMM 1447T revealed 44.0%, 44.7%, 60.1%, 48.7%, 49.1%, 60.1%, 58.9% and 60.2% relatedness respectively. The predominant quinone system is ubiquinone-9 (Q-9). The major components of the cellular fatty acids (>10%) were summed feature 8 (comprising C18:1 ω7c /C18:1 ω6c), summed feature 3 (comprising C16:1 ω7c /C16:1 ω6c) and C16:0. The detected major polar lipids were phosphatidylethanolamine (PE), phosphatidylglycerol (PG), diphosphatidylglycerol (DPG) and phosphatidylcholine (PC). On the basis of phenotypic data, chemotaxonomic and phylogenetic properties, strain AL-54T can consider as a novel species within the genus Pseudomonas, for which the name Pseudomonas lopnurensis sp. nov. is proposed. The type strain is AL-54T (= JCM 19136T = CCTCC AB 2013066T = NRRL B-59987T).

Pseudomonas populi sp. nov., an endophytic bacterium isolated from Populus euphratica.

A Gram-stain negative, aerobic, non-motile, short-rod-shaped bacterium, designated KBL-4-9T, was isolated from the storage liquid in the stems of Populus euphratica trees in Xinjiang, PR China. Strain KBL-4-9T grew at 4-45 °C (optimum 37 °C), 1-3 % (w/v) NaCl (optimum 1 %, w/v) and pH 5.5-9.5 (optimum pH 7.5). Phylogenetic analyses based on 16S rRNA gene sequences indicated that strain KBL-4-9T belonged to the genus Pseudomonas and showed the highest 16S rRNA gene sequence similarity of 97.36 % to Pseudomonas pelagia CL-AP6T, followed by Pseudomonas xinjiangensis S3-3T (97.16 %), Pseudomonas sabulinigri J64T (97.15 %) and Pseudomonas xiamenensis C10-2T (96.47 %). Analysis of strain KBL-4-9T based on the three housekeeping genes, rpoB, rpoD and gyrB, further confirmed the phylogenetic assignment of the isolates. The DNA G+C content was 61.6 mol% (sd = 2.19). DNA-DNA hybridization with P. pelagia CL-AP6T, P. xinjiangensis S3-3T and P. sabulinigri J64T revealed 49.3 % (sd = 3.04), 41.2 % and 52.5 % (sd = 4.45) relatedness, respectively. The major cellular fatty acids of strain KBL-4-9T were C16 : 0, C12 : 0, summed feature 8 (C18 : 1ω7c and/or C18 : 1ω6c) and C17 : 0 cyclo. The major isoprenoid quinone was Q-9. The major polar lipids were phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol and phosphatidylcholine. On the basis of phenotypic, chemotaxonomic and phylogenetic properties, strain KBL-4-9T is considered to represents a novel species of the genus Pseudomonas, for which the name Pseudomonas populi sp. nov. is proposed. The type strain is KBL-4-9T ( = JCM 19138T = CCTCC AB 2013069T = NRRL B-59988T).

[Microbiological characteristics of a Lysinibacillus strain isolated from Populous euphratica].

OBJECTIVE: The aim of this study was to identify the microbiological characteristics of a Lysinibacillus strain isolated from storage liquid in the stems of Populus euphratica trees. METHODS: Bacterial morphology and cultivation characteristics were studied by conventional cultivation and dyeing method. Biochemical characteristics, fatty acid components, menaquinone, polar lipids, phylogenetic analyses of 16S rRNA, determination of (G + C) mol% content and DNA- DNA hybridization were studied by polyphasic taxonomic approach. RESULTS: Strain ML-64 is Gram-positive, endospore-forming and rod-shaped. Colonies are pale-yellow, circular and entire margin. Temperature range for growth is between 10 and 45 degrees C (optimum at 37 degrees C ). The pH range for growth is between 6. 0 and 9.0 (optimum at 7.0). NaCl concentration range for growth is between 0 and 6% (optimum 3% ). Cells were positive for lipid esterase, Arginine dihydrolase, urease and Voges-Proskauer test. No sugars were fermented in the API 50CH strips. L-Serine, Methyl Pyruvate, α-Keto-Butyric, Acetoacetic Acid were oxidized. Resistant to polymyxin b (30 µg), novobiocin (30 µg), peillin G (10 U). 16S rRNA gene sequence demonstrated that strain ML-64 was closely related to Lysinibacillus chungkukjangi 2RL3-2T (100%) , Lysinibacillus sinduriensis BLB-1T (99.1%). DNA-DNA relatedness were 82% and 50. 9% with Lysinibacillus chungkukjangi 2RL3-2T and Lysinibacillus massiliensis CIP108446T, respectively. The genomic DNA G + C content of strain ML-64 was 36. 8% (mol). Major fatty acids were iso-C,,, (55. 05% ) and anteiso-C15,0 (20. 70% ). The predominant menaquinone is MK-7. Based on the phenotypic phylogenetic and genotypic analyses, the strain ML-64 is concluded to represent a new mutant strain of the Lysinibacillus chungkukjangi species, GenBank accession number is KC609752. CONCLUSION: As an endophytic bacterium of Populus euphratica, genomic structure of the strain ML-64 was greatly differentiated from the closest strain L. chungkukjangi, and suitably adapted to the endophytic environment of Populus euphratica.

Rhizobium populi sp. nov., an endophytic bacterium isolated from Populus euphratica.

An endophytic bacterium, designated K-38(T), was isolated from the storage liquid in the stems of Populus euphratica trees at the ancient Ugan River in Xinjiang, PR China. Strain K-38(T) was found to be rod-shaped, Gram-stain-negative, aerobic, non-motile and non-spore-forming. Strain K-38(T) grew at temperatures of 25-37 °C (optimum, 28 °C), at pH 6.0-9.0 (optimum, pH 7.5) and in the presence of 0-3 % (w/v) NaCl with 1 % as the optimum concentration for growth. According to phylogenetic analysis based on 16S rRNA gene sequences, strain K-38(T) was assigned to the genus Rhizobium with highest 16S rRNA gene sequence similarity of 97.2 % to Rhizobium rosettiformans W3(T), followed by Rhizobium nepotum 39/7(T) (96.5 %) and Rhizobium borbori DN316(T) (96.2 %). Phylogenetic analysis of strain K-38(T) based on the protein coding genes recA, atpD and nifH confirmed (similarities were less than 90 %) it to be a representative of a distinctly delineated species of the genus Rhizobium. The DNA G+C content was determined to be 63.5 mol%. DNA-DNA relatedness between K-38(T) and R. rosettiformans W3(T) was 48.4 %, indicating genetic separation of strain K-38(T) from the latter strain. The major components of the cellular fatty acids in strain K-38(T) were revealed to be summed feature 8 (comprising C18 : 1ω7c and/or C18 : 1ω6c; 57.2 %), C16 : 0 (13.6 %) and summed feature 2 (comprising C12 : 0 aldehyde, C14 : 0 3-OH/iso-C16 : 1 I and/or unknown ECL 10.928; 11.0 %). Polar lipids of strain K-38(T) include phosphatidylethanolamine, phosphatidylmonomethylethanolamine, phosphatidylcholine, phosphatidylglycerol, diphosphatidylglycerol, two unidentified aminophospholipids and two unidentified phospholipids. Q-10 was the major quinone in strain K-38(T). Based on phenotypic, chemotaxonomic and phylogenetic properties, strain K-38(T) represents a novel species of the genus Rhizobium, for which the name Rhizobium populi sp. nov. is proposed. The type strain is K-38(T) ( = CCTCC AB 2013068(T) = NRRL B-59990(T) = JCM 19159(T)).