The genus Caulobacter and its role in plant microbiomes.

Recent omics approaches have revealed the prevalent microbial taxa that constitute the microbiome of various plant species. Across global scales and environmental conditions, strains belonging to the bacterial genus Caulobacter have consistently been found in association with various plant species. Aligned with agroecological relevance and biotechnological advances, many scientific communications have demonstrated that several Caulobacter strains (spanning several Caulobacter species) harbor the potential to enhance plant biomass for various plant species ranging from Arabidopsis to Citrullus and Zea mays. In the past several years, co-occurrence data have driven mechanistically resolved communications about select Caulobacter-plant interactions. Given the long-standing history of Caulobacter as a model organism for cell cycle regulation, genetic studies, and the prevalence of Caulobacter species in various plant microbiomes, the genus Caulobacter offers researchers a unique opportunity to leverage for investigating plant-microbe interactions and realizing targeted biotechnological applications. In this review, recent developments regarding Caulobacter-plant interactions are presented in terms of model utility for future biotechnological investigations.

Genes related to redox and cell curvature facilitate interactions between Caulobacter strains and Arabidopsis.

Bacteria play an integral role in shaping plant growth and development. However, the genetic factors that facilitate plant-bacteria interactions remain largely unknown. Here, we demonstrated the importance of two bacterial genetic factors that facilitate the interactions between plant-growth-promoting (PGP) bacteria in the genus Caulobacter and the host plant Arabidopsis. Using homologous recombination, we disrupted the cytochrome ubiquinol oxidase (cyo) operon in both C. vibrioides CB13 and C. segnis TK0059 by knocking out the expression of cyoB (critical subunit of the cyo operon) and showed that the mutant strains were unable to enhance the growth of Arabidopsis. In addition, disruption of the cyo operon, metabolomic reconstructions, and pH measurements suggested that both elevated cyoB expression and acid production by strain CB13 contribute to the previously observed inhibition of Arabidopsis seed germination. We also showed that the crescent shape of the PGP bacterial strain C. crescentus CB15 contributes to its ability to enhance plant growth. Thus, we have identified specific genetic factors that explain how select Caulobacter strains interact with Arabidopsis plants.

Caulobacter soli sp. nov., isolated from soil sampled at Jiri Mountain, Republic of Korea.

A Gram-stain-negative, yellow-pigmented, aerobic, non-spore-forming, motile with a single polar flagellum and rod-shaped bacterium, Ji-3-8T, was isolated from a soil sample taken from Jiri Mountain, Republic of Korea. Comparative 16S rRNA gene sequence studies showed the isolate had clear affiliation with Alphaproteobacteria and the closest relatedness to Caulobacter rhizosphaerae KCTC 52515T, Caulobacter henricii ATCC 15253T, Caulobacter segnis ATCC 21756T, Caulobacter hibisci THG-AG3.4T, Caulobacter flavus RHGG3T and Caulobacter vibrioides CB51T showing 99.1, 98.9, 97.7, 97.6, 97.5 and 97.4 % 16S rRNA gene sequence similarity, respectively, and 94.7-96.5 % to the remaining species of genus Caulobacter. The predominant ubiquinone was Q-10 and the major fatty acids were C18 : 1 ω7c 11-methyl, C16 : 0, summed feature 8 (C18 : 1 ω6c and/or C18 : 1 ω7c) and summed feature 3 (C16 : 1 ω6c and/or C16 : 1 ω7c). The major polar lipids were found to be phosphatidylglycerol, two unidentified phosphoglycolipid and two unidentified glycolipids. The G+C content of the genomic DNA of strain Ji-3-8T was 68.1 mol%. Average nucleotide identity and digital DNA-DNA hybridization values of strain Ji-3-8T with C. rhizosphaerae KCTC 52515T, C. henricii ATCC 15253T, C. segnis ATCC 21756T, C. flavus RHGG3T and C. vibrioides were 79.7-87.7% and 23.0-34.3%, respectively. Based on the polyphasic evidence, it is proposed that strain Ji-3-8T forms a novel species in the genus Caulobacter, for which the name Caulobacter soli sp. nov. is proposed. The type strain is Ji-3-8T (=CCTCC AB 2019389T=KCTC 72990T).

Both clinical and environmental Caulobacter species are virulent in the Galleria mellonella infection model.

The Caulobacter genus, including the widely-studied model organism Caulobacter crescentus, has been thought to be non-pathogenic and thus proposed as a bioengineering vector for various environmental remediation and medical purposes. However, Caulobacter species have been implicated as the causative agents of several hospital-acquired infections, raising the question of whether these clinical isolates represent an emerging pathogenic species or whether Caulobacters on whole possess previously-unappreciated virulence capability. Given the proposed environmental and medical applications for C. crescentus, understanding the potential pathogenicity of this bacterium is crucial. Consequently, we sequenced a clinical Caulobacter isolate to determine if it has acquired novel virulence determinants. We found that the clinical isolate represents a new species, Caulobacter mirare that, unlike C. crescentus, grows well in standard clinical culture conditions. C. mirare phylogenetically resembles both C. crescentus and the related C. segnis, which was also thought to be non-pathogenic. The similarity to other Caulobacters and lack of obvious pathogenesis markers suggested that C. mirare is not unique amongst Caulobacters and that consequently other Caulobacters may also have the potential to be virulent. We tested this hypothesis by characterizing the ability of Caulobacters to infect the model animal host Galleria mellonella. In this context, two different lab strains of C. crescentus proved to be as pathogenic as C. mirare, while lab strains of E. coli were non-pathogenic. Further characterization showed that Caulobacter pathogenesis in the Galleria model is mediated by lipopolysaccharide (LPS), and that differences in LPS chemical composition across species could explain their differential toxicity. Taken together, our findings suggest that many Caulobacter species can be virulent in specific contexts and highlight the importance of broadening our methods for identifying and characterizing potential pathogens.

Caulobacter zeae sp. nov. and Caulobacter radicis sp. nov., novel endophytic bacteria isolated from maize root (Zea mays L.).

Four bacterial strains designated 410T, 441, 695T and 736 were isolated from maize root in Beijing, P. R. China. Based on 16S rRNA gene phylogeny, the four strains formed two clusters in the genus Caulobacter. Since strain 441 was a clonal variety of strain 410T, only three strains were selected for further taxonomic studies. The whole genome average nucleotide identity (ANI) value between strains 410T and 695T was 94.65%, and both strains shared less than 92.10% ANI values with their close phylogenetic neighbors Caulobacter vibrioides DSM 9893T, Caulobacter segnis ATCC 21756T and Caulobacter flavus CGMCC 1.15093T. Strains 410T and 695T contained Q-10 as the sole ubiquinone and their major fatty acids were C16:0, 11-methyl C18:1ω 0, 11-methyl C18: 1ω7c, summed feature 3 (C16:1ω7c and/or C16:1ω 1ω7c and/or C16: 1ω6c) and summed feature 8 (C18:1ω7c and/or C18:1ω 1ω7c and/or C18: 1ω6c). Their major polar lipids consisted of glycolipids and phosphatidylglycerol, and phenotypic tests differentiated them from their closest phylogenetic neighbors. Based on the results obtained, it is proposed that the three strains represent two novel species, for which the names Caulobacter zeae sp. nov. (type strain 410T=CGMCC 1.15991=DSM 104304) and Caulobacter radicis sp. nov. (type strain 695T=CGMCC 1.16556=DSM 106792) are proposed.

Caulobacter hibisci sp. nov., isolated from rhizosphere of Hibiscus syriacus L. (Mugunghwa flower).

A Gram-stain-negative, smooth, bright yellow-pigmented, aerobic, catalase- and oxidase-positive and rod-shaped bacterial strain was isolated from rhizosphere of Hibiscus syriacus L. (Mugunghwa flower) located in Kyung Hee University, Yongin, Gyeonggi, South Korea. Cells were dimorphic, non-motile or non-stalked, and motile by means of peritrichous flagellum. The strain, named THG-AG3.4T, grew at 15-35 °C, at pH 6.5-9.0 and in the presence of 0-1.5 % (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain THG-AG3.4T was most closely related to Caulobacter segnis ATCC 21756T (98.64 % similarity), Caulobacter vibrioides CB51T (98.57 %) and Caulobacter henricii ATCC 15253T (97.41 %). The DNA G+C content of strain THG-AG3.4T was 64.0 mol%. In DNA-DNA hybridization, the DNA-DNA relatedness between strain THG-AG3.4T and its closest phylogenetic neighbour was below 55.0 %. The predominant isoprenoid quinone detected in strain THG-AG3.4T was ubiquinone-10 (Q-10). The major polar lipids were found to be an unidentified lipid, two unidentified phosphoglycolipids, five unidentified glycolipids, eight unidentified aminolipids and phosphatidylglycerol. The major fatty acids were C16 : 0, summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c) and summed feature 8 (C18 : 1ω7c and/or C18 : 1ω6c). Thus, based on the report of the phenotypic, genotypic and phylogenetic characterization of strain THG-AG3.4T, it has been concluded that the isolate represents a novel species of the genus Caulobacter, for which the name Caulobacter hibisci sp. nov. is proposed. The type strain is THG-AG3.4T (=KACC 18849T=CCTCC AB 2016077T).

Caulobacter rhizosphaerae sp. nov., a stalked bacterium isolated from rhizosphere soil.

The Gram-reaction-negative, aerobic, white- to pale-yellow-coloured and rod-shaped bacterium with a single polar flagellum or a stalk, designated strain 7F14T, was isolated from rhizosphere soil of cultivated watermelon (Citrullus lanatus) collected from Hefei, China. Growth of strain 7F14T was observed at pH 6.0-9.0, 10-30 °C and in the presence of 0-1 % (w/v) NaCl. Cells were catalase-negative and oxidase-positive. Phylogenetic analyses based on 16S rRNA gene sequences indicated that strain 7F14T formed a phyletic lineage within the genus Caulobacter of the family Caulobacteraceae and showed the highest 16S rRNA gene sequence similarities to Caulobacter henricii ATCC 15253T (98.66 %), Caulobacter segnis ATCC 21756T (98.27 %), Caulobacter vibrioides CB51T (97.92 %) and Caulobacter flavus RHGG3T (97.44 %). The G+C content of the genomic DNA was 68.6 mol%. Strain 7F14T contained Q-10 as the sole ubiquinone and 11-methyl C18 : 1ω7c, C18 : 1ω7c, C16 : 0 and summed feature 3 (C16 : 1ω7c and/or iso-C15 : 0 2-OH) as the major fatty acids. The polar lipids profile consisted of phosphatidylglycerol, an unknown phosphoglycolipid, five unknown glycolipids, an unknown phospholipid and three unknown lipids. DNA-DNA relatedness values to the most closely related type strains Caulobacter henricii DSM 4730T and Caulobacter segnis DSM 7131T were 26.0 and 19.7 %, respectively. Based on unique phenotypic traits, and phylogenetic, chemotaxonomic and DNA-DNA hybridization results, strain 7F14T should be classified as a representative of a novel species of the genus Caulobacter, for which the name Caulobacter rhizosphaerae sp. nov. is proposed. The type strain is 7F14T (=CGMCC 1.15915T=KCTC 52515T).

Conservation of the Essential Genome Among Caulobacter and Brevundimonas Species.

When the genomes of Caulobacter isolates NA1000 and K31 were compared, numerous genome rearrangements were observed. In contrast, similar comparisons of closely related species of other bacterial genera revealed nominal rearrangements. A phylogenetic analysis of the 16S rRNA indicated that K31 is more closely related to Caulobacter henricii CB4 than to other known Caulobacters. Therefore, we sequenced the CB4 genome and compared it to all of the available Caulobacter genomes to study genome rearrangements, discern the conservation of the NA1000 essential genome, and address concerns about using 16S rRNA to group Caulobacter species. We also sequenced the novel bacteria, Brevundimonas DS20, a representative of the genus most closely related to Caulobacter and used it as part of an outgroup for phylogenetic comparisons. We expected to find that there would be fewer rearrangements when comparing more closely related Caulobacters. However, we found that relatedness was not correlated with the amount of observed "genome scrambling." We also discovered that nearly all of the essential genes previously identified for C. crescentus are present in the other Caulobacter genomes and in the Brevundimonas genomes as well. However, a few of these essential genes were only found in NA1000, and some were missing in a combination of one or more species, while other proteins were 100 % identical across species. Also, phylogenetic comparisons of highly conserved genomic regions revealed clades similar to those identified by 16S rRNA-based phylogenies, verifying that 16S rRNA sequence comparisons are a valid method for grouping Caulobacters.

Caulobacter flavus sp. nov., a stalked bacterium isolated from rhizosphere soil.

A Gram-stain-negative, aerobic, yellow-pigmented and rod-shaped bacterium with a single polar flagellum or a stalk, designated strain RHGG3T, was isolated from rhizosphere soil of cultivated watermelon (Citrullus lanatus) collected from Hefei, China. Optimal growth of strain RHGG3T was observed at pH 7.0 and 28-30 °C. Cells were catalase-positive and oxidase-negative. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain RHGG3T belonged to the genus Caulobacter and showed the highest 16S rRNA gene sequence similarities to Caulobacter segnis ATCC 21756T (98.6 %), Caulobacter vibrioides CB51T (98.3 %) and Caulobacter henricii ATCC 15253T (97.2 %). The G+C content of the genomic DNA was 70 mol%. Strain RHGG3T contained Q-10 as the sole ubiquinone and the major fatty acids (>8 %) were 11-methyl C18 : 1ω7c, C18 : 1ω7c, C16 : 0, C15 : 0 and summed feature 3 (C16 : 1ω7c and/or iso-C15 : 0 2-OH). The polar lipids were various unknown glycolipids, phosphatidylglycerol and phosphoglycolipids. DNA-DNA relatedness of strain RHGG3T to type strains of the most closely related species (Caulobacter segnis ATCC 21756T, Caulobacter vibrioides DSM 4738 and Caulobacter henricii ATCC 15253T) was 32.4-40.9 %. Based on polyphasic taxonomy analysis (phylogenetic, unique phenotypic traits, chemotaxonomic and DNA-DNA hybridizations), strain RHGG3T represents a novel species of the genus Caulobacter, for which the name Caulobacter flavus sp. nov. is proposed. The type strain is RHGG3T ( = CGMCC 1.15093T = KCTC 42581T = JCM 30763T).

Caulobacter profunda sp. nov., isolated from deep freshwater sediment.

The Gram-stain-negative, aerobic, non-spore-forming, motile, with a single polar flagellum, or non-motile (stalked) and rod-shaped bacteria, DS48-5-2(T) and DS48-6-3, were isolated from a sediment sample collected from a depth of 48 m taken from Daechung Reservoir, Republic of Korea. Comparative 16S rRNA gene sequence studies showed that the two isolates had clear affiliation with Alphaproteobacteria and the closest relatedness to Caulobacter mirabilis FWC 38(T), Caulobacter fusiformis ATCC 15257(T) and Caulobacter daechungensis H-E3-2(T) showing 98.5%, 97.3% and 97.3% 16S rRNA gene sequence similarity, respectively, and 96.1-96.7% similarity to all other species of the genus Caulobacter. The two isolates shared 100 % 16S rRNA gene sequence similarity. The predominant ubiquinone was Q-10. The major fatty acids were summed feature 8 (C18 : 1ω6c and/or C18 : 1ω7c), C16:0, C18:0ω7c 11-methyl and summed feature 3 (C16 : 1ω6c and/or C16 : 1ω7c). The G+C contents of the genomic DNA of strains DS48-5-2(T) and DS48-6-3 were 66.7 mol% and 66.2 mol%, respectively. DNA-DNA hybridization values of strains DS48-5-2(T) and DS48-6-3 with C. mirabilis FWC 38(T), C. fusiformis ATCC 15257(T) and C. daechungensis H-E3-2(T) were 19.3 %-24.4 %. Thus, based on the evidence from polyphasic studies, it is proposed that strains DS48-5-2(T) and DS48-6-3 are representatives of a novel species in the genus Caulobacter, for which the name Caulobacter profunda sp. nov. is proposed. The type strain is DS48-5-2(T) ( = KCTC 32480(T) = JCM 19440(T)).

Cellulose degradation by one mesophilic strain Caulobacter sp. FMC1 under both aerobic and anaerobic conditions.

Caulobacteria are presumed to be responsible for considerable mineralization of organic material in aquatic environments. In this study, a facultative, mesophilic and cellulolytic bacterium Caulobacter sp. FMC1 was isolated from sediments which were taken from a shallow freshwater lake and then enriched with amendment of submerged macrophyte for three months. This strain seemed to evolve a capacity to adapt redox-fluctuating environments, and could degrade cellulose both aerobically and anaerobically. Cellulose degradation percentages under aerobic and anaerobic conditions were approximately 27% and 10% after a 240-h incubation in liquid mediums containing 0.5% cellulose, respectively. Either cellulose or cellobiose alone was able to induce activities of endoglucanase, exoglucanase, and ß-1,4-glucosidase. Interestingly, ethanol was produced as the main fermentative product under anaerobic incubation on cellulose. These results could improve our understanding about cellulose-degrading process in aquatic environments, and were also useful in optimizing cellulose bioconversion process for bioethanol production.

Caulobacter daechungensis sp. nov., a stalked bacterium isolated from a eutrophic reservoir.

A Gram-stain-negative, aerobic, non-spore-forming, curved, rod-shaped bacterium, H-E3-2(T), was isolated from a water sample taken from Daechung Reservoir, Republic of Korea, during the late-blooming period of cyanobacteria. Strain H-E3-2(T) was motile with a single polar flagellum or non-motile (stalked cell). Comparative 16S rRNA gene sequence studies showed the isolate had a clear affiliation with the class Alphaproteobacteria and was most closely related to Caulobacter fusiformis ATCC 15257(T) and Caulobacter mirabilis LMG 24261(T), showing 97.6 and 97.3 % 16S rRNA gene sequence similarity, respectively, and 95.3-96.3 % similarity to all other species of the genus Caulobacter. The predominant ubiquinone was Q-10. The major fatty acids were summed feature 8 (C18 : 1ω6c and/or C18 : 1ω7c) and C16 : 0. The G+C content of the genomic DNA of strain H-E3-2(T) was 64.7 mol%. DNA-DNA hybridization values of strain H-E3-2(T) with C. fusiformis ATCC 15257(T) and C. mirabilis LMG 24261(T) were 21.2 and 19.7 %, respectively. Thus, based on the results of polyphasic analysis, it is proposed that strain H-E3-2(T) represents a novel species of the genus Caulobacter, for which the name Caulobacter daechungensis sp. nov. is proposed. The type strain is H-E3-2(T) ( = KCTC 32211(T) = JCM 18689(T)).

Codon usage methods for horizontal gene transfer detection generate an abundance of false positive and false negative results.

Bacteria acquire new DNA in a process known as horizontal gene transfer (HGT). To investigate the evolutionary impact of this transfer of DNA, various methods have been developed to detect past HGT events. For example, codon usage-based methods detect the presence of transferred genes by identifying atypical patterns of codon usage. However, some inherited genes exhibit atypical codon usage and some transferred genes have codon usage patterns similar to those of the inherited genes. In this study, we used a comparative phylogenetic approach with Methylobacterium and Caulobacter species to demonstrate that even well-designed codon usage methods fail to detect many HGT events and generate a high rate of false positives (60-75 %) and false negatives (23-61 %). Therefore, we recommend caution when employing codon usage methods to identify transferred genes and suggest that the rapidly increasing availability of bacterial genome sequences makes the phylogenetic approach the method of choice.

Reclassification and emended description of Caulobacter leidyi as Sphingomonas leidyi comb. nov., and emendation of the genus Sphingomonas.

'Caulobacter leidyi' DSM 4733(T) has been shown to be affiliated with the family Sphingomonadaceae instead of the Caulobacteraceae, and due to its poor characterization has been omitted from the current edition of Bergey's Manual of Systematic Bacteriology and removed to limbo. We isolated a novel sphingoglycolipid-containing dimorphic prosthecate bacterium, designated strain 247, from a pre-alpine freshwater lake. Strain 247 and 'Caulobacter leidyi' DSM 4733(T) were characterized in detail. The rod-shaped cells were Gram-stain-negative, aerobic, catalase- and oxidase-positive, and formed a stalk or polar flagellum. Both strains grew optimally at 28-30 °C, and pH 6.0-8.0. The major fatty acids were C(18 : 1)ω7c, C(16 : 0) and 11-methyl C(18 : 1)ω7c. C(14 : 0) 2-OH represents the major 2-hydroxy fatty acid. Q-10 was the major respiratory quinone and the major polar lipids were diphosphatidylglycerol, phosphatidyldimethylethanolamine, phosphatidylglycerol, phosphatidylmonomethylethanolamine, phosphatidylethanolamine, phosphatidylcholine, three glycolipids, two phosphoaminolipids and two unidentified sphingoglycolipids. The major polyamine was sym-homospermidine. The G+C content of genomic DNA of strains 247 and DSM 4733(T) was 67.6 mol% and 67.0 mol%, respectively. According to 16S rRNA gene sequence analysis and DNA-DNA hybridization, strains DSM 4733(T) and 247 were phylogenetically closely related (99.6 % 16S rRNA gene sequence similarity, 82.9 % DNA-DNA hybridization value) and affiliated to the genus Sphingomonas. The closest recognized species was Sphingomonas aquatilis DSM 15581(T) (98.1 % sequence similarity). In addition, the presence of cystine arylamidase, absence of ß-galactosidase, and the inability to utilize l-arabinose, galactose and sucrose distinguished strains DSM 4733(T) and 247 from most other members of the family Sphingomonadaceae. So far, the dimorphic life cycle that involves a prosthecate and a flagellated stage is unique for strains DSM 4733(T) and 247 among all members of the family Sphingomonadaceae. Therefore, Caulobacter leidyi is reclassified as Sphingomonas leidyi, with the type strain DSM 4733(T) ( = ATCC 15260(T) = CIP 106443(T) = VKM B-1368(T)) and strain 247 (DSM 25078 = LMG 26658) as an additional strain of this species.

Novel root nodule bacteria belonging to the genus Caulobacter.

AIMS: Aim of this study is to determine the genetic variation of rhizobia associated with horse gram [Macrotyloma uniflorum (Lam.) Verdc.] plants grown in different regions of Andhra Pradesh, India. METHODS AND RESULTS: Four representative isolates having most representative characters from the previous characterization were selected for 16S rRNA sequence. The sequences were submitted to the NCBI GenBank and Ribosomal Database Project (RDP). The isolates HGR-4, 6 and 13 showed more than 99% homology between them and they were grouped with Rhizobium reference strains where as the isolate HGR-25 showed 87.1, 87.4 and 87.2% homology with the isolates HGR-4, 6 and 13, respectively, and were grouped with reference strains for Caulobacter. The nodulation ability of these isolates on horse gram was confirmed by inoculation tests. CONCLUSIONS: The isolate HGR-25 was identified as Caulobacter isolated from the plants growing in soil samples collected from Khareemnagar district, Andhra Pradesh, India. Inoculation tests revealed that Caulobacter formed nodules on horse gram. It was also confirmed by RDP. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report that a legume was nodulated by a member of the genus Caulobacter, which belongs to the family Caulobacteriaceae in the order Caulobacterales of Alphaproteobacteria.

Efficacy of various chemical disinfectants on biofilms formed in spacecraft potable water system components.

As the provision of potable water is critical for successful habitation of the International Space Station (ISS), life support systems were installed in December 2008 to recycle both humidity from the atmosphere and urine to conserve available water in the Station. In-flight pre-consumption testing from the dispensing needle at the Potable Water Dispenser (PWD) indicated that bacterial concentrations exceeded the current ISS specifications of 50 colony-forming units (CFU) ml(-1). Subsequent investigations revealed that a corrugated stainless steel flex hose upstream of the dispensing needle in the PWD was filled with nonsterile water and left at room temperature for more than 1 month before launch. To simulate biofilm formation that was suspected in the flight system, sterile flex hoses were seeded with a consortium of bacterial isolates previously recovered from other ISS water systems, including Ralstonia pickettii, Burkholderia multivorans, Caulobacter vibrioides, and Cupriavidus pauculus. After incubation for 5 days, the hoses were challenged with various chemical disinfectants including hydrogen peroxide (H2O2), colloidal silver, and buffered pH solutions to determine the ability of the disinfectants to decrease and maintain bacterial concentrations below ISS specifications. The disinfection efficacy over time was measured by collecting daily heterotrophic plate counts after exposure to the disinfectants. A single flush with either 6% H2O2 solution or a mixture of 3% H2O2 and 400 ppb colloidal silver effectively reduced the bacterial concentrations to <1 CFU ml(-1) for a period of up to 3 months.

Caulobacter ginsengisoli sp. nov., a novel stalked bacterium isolated from ginseng cultivating soil.

A Gram negative, aerobic, nonspore-forming, straight or curved rod-shaped bacterium, designated Gsoil 317T, was isolated from soil of a ginseng field in Pocheon Province (South Korea) and was characterized using a polyphasic approach. Cells were dimorphic, with stalk (or prostheca) and nonmotile or nonstalked and motile, by means of a single polar flagellum. Comparative analysis of 16S rRNA gene sequences revealed that strain Gsoil 317T was most closely related to Caulobacter mirabilis LMG 24261T (97.2%), Caulobacter fusiformis ATCC 15257T (97.1%), Caulobacter segnis LMG 17158T (97.0%), Caulobacter vibrioides DSM 9893T (96.8%), and Caulobacter henricii ATCC 15253T (96.7%). The sequence similarities to any other recognized species within Alphaproteobacteria were less than 96.0%. The detection of Q-10 as the major respiratory quinone and a fatty acid profile with summed feature 7 (C18:1 omega7c and/or C18:1 omega9t and/or C18:1 omega12t; 56.6%) and C16:0 (15.9%) as the major fatty acids supported the affiliation of strain Gsoil 317T to the genus Caulobacter. The G+C content of the genomic DNA was 65.5 mol%. DNA-DNA hybridization experiments showed that the DNA-DNA relatedness values between strain Gsoil 317T and its closest phylogenetic neighbors were below 11%. On the basis of its phenotypic properties and phylogenetic distinctiveness, strain Gsoil 317T should be classified as representing a novel species in the genus Caulobacter, for which the name Caulobacter ginsengisoli sp. nov. is proposed. The type strain is Gsoil 317T (=KCTC 12788T= DSM 18695T).

Phylogeny by a polyphasic approach of the order Caulobacterales, proposal of Caulobacter mirabilis sp. nov., Phenylobacterium haematophilum sp. nov. and Phenylobacterium conjunctum sp. nov., and emendation of the genus Phenylobacterium.

Three strains of Gram-negative, rod-shaped, non-spore-forming bacteria were isolated from fresh water and human blood. As determined by analyses of 16S rRNA gene sequences, the prosthecate strain FWC 38T was affiliated to the alphaproteobacterial genus Caulobacter, with Caulobacter henricii (96.8 %) and Caulobacter fusiformis (96.8 %) as its closest relatives. The non-prosthecate strain LMG 11050T and the prosthecate strain FWC 21T both belonged to the genus Phenylobacterium with Phenylobacterium koreense (96.9 %) and Phenylobacterium immobile (96.3 %) as the closest relatives. This affiliation was supported by chemotaxonomic data (polar lipids and cellular fatty acids). Physiological and biochemical tests allowed genotypic and phenotypic differentiation of the novel strains from all hitherto recognized species of the genera Caulobacter and Phenylobacterium. The strains therefore represent novel species, for which the names Caulobacter mirabilis sp. nov. (type strain FWC 38T=LMG 24261T=CCUG 55073T), Phenylobacterium conjunctum (type strain FWC 21T=LMG 24262T=CCUG 55074T), the first described prosthecate Phenylobacterium species, and Phenylobacterium haematophilum sp. nov. (type strain LMG 11050T=CCUG 26751T) are proposed. Marker nucleotides within the 16S rRNA genes were determined for the genera Asticcacaulis, Brevundimonas, Caulobacter and Phenylobacterium and the description of the genus Phenylobacterium is emended.

Biological filtration limits carbon availability and affects downstream biofilm formation and community structure.

Carbon removal strategies have gained popularity in the mitigation of biofouling in water reuse processes, but current biofilm-monitoring practices based on organic-carbon concentrations may not provide an accurate representation of the in situ biofilm problem. This study evaluated a submerged microtiter plate assay for direct and rapid monitoring of biofilm formation by subjecting the plates to a continuous flow of either secondary effluent (SE) or biofilter-treated secondary effluent (BF). This method was very robust, based on a high correlation (R(2) = 0.92) between the biomass (given by the A(600) in the microtiter plate assay) and the biovolume (determined from independent biofilms developed on glass slides under identical conditions) measurements, and revealed that the biomasses in BF biofilms were consistently lower than those in SE biofilms. The influence of the organic-carbon content on the biofilm community composition and succession was further evaluated using molecular tools. Terminal restriction fragment length polymorphism analysis of 16S rRNA genes revealed a group of pioneer colonizers, possibly represented by Sphingomonadaceae and Caulobacter organisms, to be common in both SE and BF biofilms. However, differences in organic-carbon availabilities in the two water samples eventually led to the selection of distinct biofilm communities. Alphaproteobacterial populations were confirmed by fluorescence in situ hybridization to be enriched in SE biofilms, while Betaproteobacteria were dominant in BF biofilms. Cloning analyses further demonstrated that microorganisms adapted for survival under low-substrate conditions (e.g., Aquabacterium, Caulobacter, and Legionella) were preferentially selected in the BF biofilm, suggesting that carbon limitation strategies may not achieve adequate biofouling control in the long run.