The class Alphaproteobacteria is comprised of a diverse assemblage of Gram-negative bacteria that includes organisms of varying morphologies, physiologies and habitat preferences many of which are of clinical and ecological importance. Alphaproteobacteria classification has proved to be difficult, not least when taxonomic decisions rested heavily on a limited number of phenotypic features and interpretation of poorly resolved 16S rRNA gene trees. Despite progress in recent years regarding the classification of bacteria assigned to the class, there remains a need to further clarify taxonomic relationships. Here, draft genome sequences of a collection of genomes of more than 1000 Alphaproteobacteria and outgroup type strains were used to infer phylogenetic trees from genome-scale data using the principles drawn from phylogenetic systematics. The majority of taxa were found to be monophyletic but several orders, families and genera, including taxa recognized as problematic long ago but also quite recent taxa, as well as a few species were shown to be in need of revision. According proposals are made for the recognition of new orders, families and genera, as well as the transfer of a variety of species to other genera and of a variety of genera to other families. In addition, emended descriptions are given for many species mainly involving information on DNA G+C content and (approximate) genome size, both of which are confirmed as valuable taxonomic markers. Similarly, analysis of the gene content was shown to provide valuable taxonomic insights in the class. Significant incongruities between 16S rRNA gene and whole genome trees were not found in the class. The incongruities that became obvious when comparing the results of the present study with existing classifications appeared to be caused mainly by insufficiently resolved 16S rRNA gene trees or incomplete taxon sampling. Another probable cause of misclassifications in the past is the partially low overall fit of phenotypic characters to the sequence-based tree. Even though a significant degree of phylogenetic conservation was detected in all characters investigated, the overall fit to the tree varied considerably.

/pdf/analysis-of-1-000-type-strain-genomes-substantially-improves-57hicbb77a.pdf

Analysis of 1,000+ Type-Strain Genomes Substantially Improves Taxonomic Classification of Alphaproteobacteria.

Role of circular RNAs in brain development and CNS diseases.

A polyphasic study was carried out to clarify the taxonomic position of a pink-coloured, Gram-negative bacterium isolated from air in the Suwon region of Korea. The novel strain, 5420S-30(T), grew in the temperature range of 5-35 degrees C (optimum 30 degrees C) and pH range of 5-9 (optimum pH 6-7). Growth did not occur in the presence of as little as 1 % (w/v) NaCl. It contained Q-10 as the predominant ubiquinone. The major fatty acids were C(18 : 1)omega7c, summed feature 3 (C(16 : 1)omega7c/iso-C(15 : 0) 2-OH) and C(16 : 0) (together representing 77.7 % of the total). The DNA G+C content was 72.1 mol%. The 16S rRNA gene sequence similarities between strain 5420S-30(T) and members of the genus Roseomonas were in the range 91.6-95.1 %, with the highest sequence similarities (95.1 and 94.5 %, respectively) to Roseomonas aquatica TR53(T) and Roseomonas gilardii ATCC 49956(T). On the basis of its phenotypic and genotypic characteristics, strain 5420S-30(T) represents a novel species of the genus Roseomonas, for which the name Roseomonas aerilata sp. nov. is proposed, with 5420S-30(T) (=KACC 12521(T) =DSM 19363(T)) as the type strain.

Roseomonas aerilata sp. nov., isolated from an air sample. Int J Syst Evol Microbiol

Thiobencarb is a thiocarbamate herbicide used in rice paddies worldwide. Microbial degradation plays a crucial role in the dissipation of thiobencarb in the environment. However, the physiological and genetic mechanisms underlying thiobencarb degradation remain unknown. In this study, a novel thiobencarb degradation pathway was proposed in Acidovorax sp. strain T1. Thiobencarb was oxidized and cleaved at the C-S bond, generating diethylcarbamothioic S-acid and 4-chlorobenzaldehyde (4CDA). 4CDA was then oxidized to 4-chlorobenzoic acid (4CBA) and hydrolytically dechlorinated to 4-hydroxybenzoic acid (4HBA). The identification of catabolic genes suggested further hydroxylation to protocatechuic acid (PCA) and finally degradation through the protocatechuate 4,5-dioxygenase pathway. A novel two-component monooxygenase system identified in the strain, TmoAB, was responsible for the initial catabolic reaction. TmoA shared 28 to 32% identity with the oxygenase components of pyrimidine monooxygenase from Agrobacterium fabrum, alkanesulfonate monooxygenase from Pseudomonas savastanoi, and dibenzothiophene monooxygenase from Rhodococcus sp. TmoB shared 25 to 37% identity with reported flavin reductases and oxidized NADH but not NADPH. TmoAB is a flavin mononucleotide (FMN)-dependent monooxygenase and catalyzed the C-S bond cleavage of thiobencarb. Introduction of tmoAB into cells of the thiobencarb degradation-deficient mutant T1m restored its ability to degrade and utilize thiobencarb. A dehydrogenase gene, tmoC, was located 7,129 bp downstream of tmoAB, and its transcription was clearly induced by thiobencarb. The purified TmoC catalyzed the dehydrogenation of 4CDA to 4CBA using NAD+ as a cofactor. A gene cluster responsible for the complete 4CBA metabolic pathway was also cloned, and its involvement in thiobencarb degradation was preliminarily verified by transcriptional analysis.IMPORTANCE Microbial degradation is the main factor in thiobencarb dissipation in soil. In previous studies, thiobencarb was degraded initially via N-deethylation, sulfoxidation, hydroxylation, and dechlorination. However, enzymes and genes involved in the microbial degradation of thiobencarb have not been studied. This study revealed a new thiobencarb degradation pathway in Acidovorax sp. strain T1 and identified a novel two-component FMN-dependent monooxygenase system, TmoAB. Under TmoAB-mediated catalysis, thiobencarb was cleaved at the C-S bond, producing diethylcarbamothioic S-acid and 4CDA. Furthermore, the downstream degradation pathway of thiobencarb was proposed. Our study provides the physiological, biochemical, and genetic foundation of thiobencarb degradation in this microorganism.

A Novel Aerobic Degradation Pathway for Thiobencarb Is Initiated by the TmoAB Two-Component Flavin Mononucleotide-Dependent Monooxygenase System in Acidovorax sp. Strain T1.

A Gram-stain-negative, facultative anaerobic, non-motile and rod-shaped bacterial strain, designated LX32T, was isolated from arsenic and cadmium contaminated farmland soil. Phylogenetic analysis based on 16S rRNA gene sequence indicated that strain LX32T was closely related to 
Phenylobacterium hankyongense
HKS-05T (97.7 % sequence similarity), 
Phenylobacterium kunshanense
CCTCC AB 2013085T (97.4 %) and 
Phenylobacterium deserti
CCTCC AB 2016297T (97.1 %). The average nucleotide identity values of the whole genome sequences of LX32T/P. 
hankyongense
HKS-05T, LX32T/P. 
kunshanense
CCTCC AB 2013085T and LX32T/P. 
deserti
CCTCC AB 2016297T were 79.8, 77.9 and 77.5 %, respectively. Its genome size was 4.02 Mb, comprising 3998 predicted genes with a DNA G+C content of 70.1 mol%. The major fatty acids were C15 : 0, C16 : 0 and summed feature 8 (comprising C18 : 1ω7c and/or C18 : 1ω6c). The polar lipid profiles consisted of phosphatidylglycerol, aminophospholipid, seven glycolipids and two unidentified polar lipids. The predominantly respiratory quinone was ubiquinone-10. Based on polyphasic analyses, the isolate is considered to represent a novel species of the genus 
Phenylobacterium
, for which the name 
Phenylobacterium
soli sp. nov. is proposed. The type strain is LX32T (=KCTC 62522=CCTCC AB 2018055).

Phenylobacterium soli sp. nov., isolated from arsenic and cadmium contaminated farmland soil.

A novel aerobic, Gram-stain-negative, motile bacterium, designated strain BUT-10T, was isolated from the sludge of a pesticide manufacturing factory in Kunshan, China. Cells were rod-shaped (0.4–0.45×0.9–1.4 µm) and colonies were white, circular with entire edges and had a smooth surface. The strain grew at 25–37 °C, at pH 6.0–8.0 and with 0–0.5 % NaCl. Phylogenetic analysis based on 16S rRNA gene sequence comparisons revealed that strain BUT-10T was a member of the genus 
Phenylobacterium
, and showed highest sequence similarities to 
Phenylobacterium muchangponense
A8T (97.49 %), 
Phenylobacterium immobile
DSM 1986T (97.14 %) and 
Phenylobacterium lituiforme
FaiI3T (96.34 %). Major fatty acids (>5 %) were summed feature 8 (comprising C18 : 1ω7c and/or C18 : 1ω6c), C16 : 0 and summed feature 3 (comprising C16 : 1ω7c and/or C16 : 1ω6c). The major isoprenoid quinone was ubiquinone-10. The DNA G+C content was 71.85 mol%. Strain BUT-10T showed low DNA–DNA relatedness with 
P. muchangponense
A8T (15.7±2.9 %) and 
P. immobile
DSM 1986T (12.8±1.1 %). On the basis of the phenotypic, phylogenetic and genotypic data, strain BUT-10T is considered to represent a novel species of the genus 
Phenylobacterium
, for which the name Phenylobacterium kunshanense sp. nov. is proposed. The type strain is BUT-10T ( = CCTCC AB 2013085T = KCTC 42014T).

Phenylobacterium kunshanense sp. nov., isolated from the sludge of a pesticide manufacturing factory.

A Gram-stain-negative, aerobic, short rod-shaped, non-endospore-forming, ivory-pigmented and non-motile bacterium, designated strain BUT-5T, was isolated from activated sludge of an herbicides-manufacturing wastewater treatment facility in Jiangsu Province, China. The major fatty acids (>5 % of total fatty acids) were C16 : 0, C18 : 1 2-OH and summed feature 8 (C18 : 1ω7c and/or C18 : 1ω6c). The predominant respiratory quinone was ubiquinone Q-10. The polar lipids profile of strain BUT-5T included diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine and two unknown aminolipids. The DNA G+C content was 67.6 mol%. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain BUT-5T showed the highest sequence similarities to Roseomonas soli 5N26T (97.5 % 16S rRNA gene sequence similarity), followed by Roseomonas lacus TH-G33T (97.3 %) and Roseomonas terrae DS-48T (97.1 %). Strain BUT-5T showed low DNA-DNA relatedness with Roseomonas soli KACC 16376T (41 %), Roseomonas lacus KACC 11678T (46 %) and Roseomonas terrae KACC 12677T (42 %), respectively. On the basis of phenotypic and genotypic properties, as well as chemotaxonomic data, strain BUT-5T represents a novel species of the genus Roseomonas, for which the name Roseomonas eburnea sp. nov. is proposed. The type strain is BUT-5T ( = CCTCC AB2013276T = KACC 17166T).

Roseomonas eburnea sp. nov., isolated from activated sludge

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Phenylobacterium kunshanense sp. nov., isolated from the sludge of a pesticide manufacturing factory.

Roseomonas eburnea sp. nov., isolated from activated sludge