recA Gene Research Articles

Biodiversity and Geographic Distribution of Rhizobia Nodulating With Vigna minima.

Vigna minima is a climbing annual plant widely distributed in barren wilderness, grass land, and shrub bush of China and other countries such as Japan. However, the rhizobia nodulating with this plant has never been systematically studied. In order to reveal the biodiversity of nodulating rhizobia symbiosis with V. minima, a total of 874 rhizobium isolates were obtained from root nodules of the plant spread in 11 sampling sites of Shandong Peninsula, China, and they were designated as 41 haplotypes in the genus Bradyrhizobium based upon recA sequence analyses. By multilocus sequence analysis (MLSA) of five housekeeping genes (dnaK, glnII, gyrB, recA, and rpoB), the 41 strains representing different recA haplotypes were classified into nine defined species and nine novel genospecies. Bradyrhizobium elkanii, Bradyrhizobium ferriligni, and Bradyrhizobium pachyrhizi were the predominant and universally distributed groups. The phylogeny of symbiotic genes of nodC and nifH showed similar topology and phylogenetic relationships, in which all the representative strains were classified into two clades grouped with strains nodulating with Vigna spp., demonstrating that Vigna spp. shared common nodulating groups in the natural environment. All the representative strains formed nodules with V. minima in a nodulation test performed in green house conditions. The correlation between V. minima nodulating rhizobia and soil characteristics analyzed by CANOCO indicates that available nitrogen, total nitrogen, and organic carbon in the soil samples were the main factors affecting the distribution of rhizobia isolated in this study. This study systematically uncovered the biodiversity and distribution characteristics of V. minima nodulating rhizobia for the first time, which provided novel information for the formation of the corresponding rhizobium community.

Decreasing the Cut-off Score Value of MALDI-ToF MS Increase the Identities of Burkholderia cepacia Complex Species.

Burkholderia cepacia complex (Bcc) comprises 24 related species genetically distinct, associated with high mortality in cystic fibrosis (CF) patients. Due to a high level of similarity among Bcc species, accurate identification has been problematic, and most conventional and automated phenotypic tests have shown low accuracy. We evaluated accuracy of MALDI-ToF MS decreasing the cut-off score value to distinguish Bcc species compared to recA gene sequencing. A total of 145 Bcc isolates were analyzed. B. vietnamiensis (41.37%), B. cenocepacia IIIA (23.44%), B. multivorans (20%), B. cenocepacia IIIB (11.03%), and B. contaminans (2.75%) among other species were identified by recA sequencing. MALDI-ToF MS identified 100% of Bcc isolates at the genus level and 53.1% at the species level. By decreasing cut-off values for ≥1.70, the correct identification at the species level increased to 74.5%. MALDI-ToF MS proved to be useful at the genus level identification, but it still requires improvements that allow more precise identification, requiring continuous updates and addition of new spectra to its database. A review of interpretative criteria is a field to be explored with a large collection of Bcc species.

Paraburkholderia spp. are the main rhizobial microsymbionts of Mimosa tenuiflora (Willd.) Poir. in soils of the Brazilian tropical dry forests (Caatinga biome)

Mimosa tenuiflora (Willd.) Poir. is widespread in southern and central American drylands, but little information is available concerning its associated rhizobia. Therefore, this study aimed to characterize M. tenuiflora rhizobia from soils of the tropical dry forests (Caatinga) in Pernambuco State, Brazil, at the molecular and symbiotic levels. Soil samples of pristine Caatinga areas in four municipalities were used to grow M. tenuiflora. First, the bacteria from root nodules were subjected to nodC/nifH gene amplification, and the bacteria positive for both genes had the 16S rRNA gene sequenced. Then, ten strains were evaluated using recA, gyrB, and nodC gene sequences, and seven of them had their symbiotic efficiency assessed. Thirty-two strains were obtained and 22 of them were nodC/nifH positive. Twenty strains clustered within Paraburkholderia and two within Rhizobium by 16S rRNA gene sequencing. The beta-rhizobia were similar to P. phenoliruptrix (12) and P. diazotrophica (8). Both alpha-rhizobia were closely related to R. miluonense. The recA + gyrB phylogenetic analysis clustered four and five strains within the P. phenoliruptrix and P. diazotrophica branches, respectively, but they were somewhat divergent to the 16S rRNA phylogeny. For Rhizobium sp. ESA 637, the recA + gyrB phylogeny clustered the strain with R. jaguaris. The nodC phylogeny indicated that ESA 626, ESA 629, and ESA 630 probably represented a new symbiovar branch. The inoculation assay showed high symbiotic efficiency for all tested strains. The results indicated high genetic diversity and efficiency of M. tenuiflora rhizobia in Brazilian drylands and included P. phenoliruptrix-like bacteria in the list of efficient beta-rhizobia in the Caatinga biome.

Genomics reveals the novel species placement of industrial contaminant isolates incorrectly identified as Burkholderia lata.

The Burkholderia cepacia complex (Bcc) is a closely related group of bacteria, composed of at least 20 different species, the accurate identification of which is essential in the context of infectious diseases. In industry, they can contaminate non-food products, including home and personal care products and cosmetics. The Bcc are problematic contaminants due to their ubiquitous presence and intrinsic antimicrobial resistance, which enables them to occasionally overcome preservation systems in non-sterile products. Burkholderia lata and Burkholderia contaminans are amongst the Bcc bacteria encountered most frequently as industrial contaminants, but their identification is not straightforward. Both species were historically established as a part of a group known collectively as taxon K, based upon analysis of the recA gene and multilocus sequence typing (MLST). Here, we deploy a straightforward genomics-based workflow for accurate Bcc classification using average nucleotide identity (ANI) and core-gene analysis. The workflow was used to examine a panel of 23 Burkholderia taxon K industrial strains, which, based on MLST, comprised 13 B. lata, 4 B. contaminans and 6 unclassified Bcc strains. Our genomic identification showed that the B. contaminans strains retained their classification, whilst the remaining strains were reclassified as Burkholderia aenigmatica sp. nov. Incorrect taxonomic identification of industrial contaminants is a problematic issue. Application and testing of our genomic workflow allowed the correct classification of 23 Bcc industrial strains, and also indicated that B. aenigmatica sp. nov. may have greater importance than B. lata as a contaminant species. Our study illustrates how the non-food manufacturing industry can harness whole-genome sequencing to better understand antimicrobial-resistant bacteria affecting their products.

PprA Protein Inhibits DNA Strand Exchange and ATP Hydrolysis of Deinococcus RecA and Regulates the Recombination in Gamma-Irradiated Cells.

DrRecA and PprA proteins function are crucial for the extraordinary resistance to γ-radiation and DNA strand break repair in Deinococcus radiodurans. DrRecA mediated homologous recombination help in DNA strand break repair and cell survival, while the PprA protein confers radio-resistance via its roles in DNA repair, genome maintenance, and cell division. Genetically recA and pprA genes interact and constitute an epistatic group however, the mechanism underlying their functional interaction is not clear. Here, we showed the physical and functional interaction of DrRecA and PprA protein both in solution and inside the cells. The absence of the pprA gene increases the recombination frequency in gamma-irradiated D. radiodurans cells and genomic instability in cells growing under normal conditions. PprA negatively regulates the DrRecA functions by inhibiting DrRecA mediated DNA strand exchange and ATPase function in vitro. Furthermore, it is shown that the inhibitory effect of PprA on DrRecA catalyzed DNA strand exchange was not due to sequestration of homologous dsDNA and was dependent on PprA oligomerization and DNA binding property. Together, results suggest that PprA is a new member of recombination mediator proteins (RMPs), and able to regulate the DrRecA function in γ-irradiated cells by protecting the D. radiodurans genome from hyper-recombination and associated negative effects.

Characterization of Retama sphaerocarpa microsymbionts in Zaida lead mine tailings in the Moroccan middle Atlas

In the Moroccan Middle Atlas, the tailings rich in lead and other metal residues, in the abandoned Zaida mining district, represent a real threat to environment and the neighboring villages’ inhabitants’ health. In this semi-arid to arid area, phytostabilisation would be the best choice to limit the transfer of heavy metals to populations and groundwater. The aim of this work was to characterize the bacteria that nodulate Retama sphaerocarpa, spontaneous nitrogen fixing shrubby legume, native to the Zaida mining area, with great potential to develop for phytostabilisation. Forty-three bacteria isolated from root nodules of the plant were characterized. Based on REP-PCR and ARDRA, four strains were selected for further molecular analyzes. The 16S rRNA gene sequences analysis revealed that the isolated strains are members of the genus Bradyrhizobium, and the phylogenetic analysis of the housekeeping genes glnII, atpD, gyrB, rpoB, recA and dnaK individual sequences and their concatenation showed that the strains are close to B. algeriense RST89T and B. valentinum LmjM3T with similarity percentages of 89.07% to 95.66% which suggest that the newly isolated strains from this mining site may belong to a potential novel species. The phylogeny of the nodA and nodC genes showed that the strains belong to the symbiovar retamae of the genus Bradyrhizobium. These strains nodulate also R. monosperma, R. dasycarpa and Lupinus luteus.

Inactivation of UmuC Protein Significantly Reduces Resistance to Ciprofloxacin and SOS Mutagenesis in Escherichia coli Mutants Harboring Intact umuD Gene

Background: Ciprofloxacin induces SOS response and mutagenesis by activation of UmuD’2C (DNA polymerase V) and DinB (DNA polymerase IV) in Escherichia coli, leading to antibiotic resistance during therapy. Inactivation of DNA polymerase V can result in the inhibition of mutagenesis in E. coli. Objectives: The aim of this research was to investigate the effect of UmuC inactivation on resistance to ciprofloxacin and SOS mutagenesis in E. coli mutants. Methods: Ciprofloxacin-resistant mutants were produced in a umuC- genetic background in the presence of increasing concentrations of ciprofloxacin. The minimum inhibitory concentration of umuC-mutants was measured by broth dilution method. Alterations in the rifampin resistance-determing region of rpoB gene were assessed by PCR amplification and DNA sequencing. The expression of SOS genes was measured by quantitative real-time PCR assay. Results: Results showed that despite the induction of SOS response (overexpression of recA, dinB, and umuD genes) following exposure to ciprofloxacin in E. coli umuC mutants, resistance to ciprofloxacin and SOS mutagenesis significantly decreased. However, rifampicin-resistant clones emerged in this genetic background. One of these clones showed mutations in the rifampicin resistance-determining region of rpoB (cluster II). The low mutation frequency of E. coli might be associated with the presence and overexpression of umuD gene, which could somehow limit the activity of DinB, the location and type of mutations in the β subunit of RNA polymerase. Conclusions: In conclusion, for increasing the efficiency of ciprofloxacin against Gram-negative bacteria, use of an inhibitor of umuC, along with ciprofloxacin, would be helpful.

Characterization of the Recombinant Glutamate Decarboxylase of Lactobacillus brevis G144 Isolated from Galchi Jeotgal, a Korean Salted and Fermented Seafood

A γ-aminobutyric acid (GABA)-producing microorganism was isolated from galchi (hairtail fish, Trichiurus lepturus) jeotgal, a Korean salted and fermented seafood. The G144 isolate produced GABA excessively when incubated in MRS broth containing monosodium glutamate (MSG, 3%, w/v). G144 was identified as Lactobacillus brevis through 16S rRNA and recA gene sequencing. gadB and gadC encoding glutamate decarboxylase (GAD) and glutamate/GABA antiporter, respectively, were cloned and gadB was located downstream of gadC. The operon structure of gadCB was confirmed by reverse transcription (RT)-polymerase chain reaction. gadB was overexpressed in Escherichia coli and recombinant GAD was purified and its size was 54.4 kDa as evidenced by SDS-PAGE results. Maximum GAD activity was observed at pH 5.0 and 40℃ and the activity was dependent on pyridoxal 5''-phophate. The Km and Vmax of GAD were 8.6 mM and 0.01 mM/min, respectively.

The endemic Chamaecytisus albidus is nodulated by symbiovar genistearum of Bradyrhizobium in the Moroccan Maamora Forest

Out of 54 isolates from root nodules of the Moroccan-endemic Chamaecytisus albidus plants growing in soils from the Maamora cork oak forest, 44 isolates formed nodules when used to infect their original host plant. A phenotypic analysis showed the metabolic diversity of the strains that used different carbohydrates and amino acids as sole carbon and nitrogen sources. The isolates grew on media with pH values ranging from 6 to 8. However, they did not tolerate high temperatures or drought and they did not grow on media with salt concentrations higher than 85 mM. REP-PCR fingerprinting grouped the strains into 12 clusters, of which representative strains were selected for ARDRA and rrs analyses. The rrs gene sequence analysis indicated that all 12 strains were members of the genus Bradyrhizobium and their phylogeny showed that they were grouped into two different clusters. Two strains from each group were selected for multilocus sequence analysis (MLSA) using atpD, recA, gyrB and glnII housekeeping genes. The inferred phylogenetic trees confirmed that the strains clustered into two divergent clusters. Strains CM55 and CM57 were affiliated to the B. canariense/B. lupini group, whereas strains CM61 and CM64 were regrouped within the B. cytisi/B. rifense lineage. The analysis of the nodC symbiotic gene affiliated the strains to the symbiovar genistearum. The strains were also able to nodulate Retama monosperma, Lupinus luteus and Cytisus monspessulanus, but not Phaseolus vulgaris or Glycine max.Inoculation tests with C. albidus showed that some strains could be exploited as efficient inocula that could be used to improve plant growth in the Maamora forest.

Streptomyces polyasparticus sp. nov. isolated from cotton field soil by a medium applied with polyaspartic acid.

A novel Streptomyces strain (TRM66268-LWLT) was isolated from cotton field soil by a medium supplied with polyaspartic acid (PASP) at Alar, Xinjiang, Northwest PR China, and characterized using a polyphasic taxonomic approach. The strain was found to degrade PASP, and grow well on the medium to take PASP as the sole carbon source. The TRM66268-LWLT fermentation broth was applied to the surface of PASP, and there were pores on the surface of PASP after a period of time. The strain was observed to be Gram-stain-positive and to form greyish-white aerial mycelia that differentiated into straight spore chains with round spores. The whole-cell sugar pattern of TRM 66268-LWLT consisted of ribose, mannose and arabinose, and the principal phospholipids were found to be diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol mannoside, phosphatidylinositol and two undetermined polar lipids. The predominant menaquinones were MK-7, MK-7(H4), MK-9(H8), MK-10(H6). The diagnostic cell wall amino acid was identified as LL-diaminopimelic acid. The G+C content of strain TRM66268-LWLT was 70.11mol%. The average nucleotide identity value between strain TRM66268-LWLT and the phylogenetically related strain Streptomyces indicus IH32-1T was calculated to be 85.49%. The digital DNA-DNA hybridization value between them was 30.40%. A multilocus sequence analysis of five house-keeping genes (atpD, gyrB, rpoB, recA and trpB) also illustrated that strain TRM66268-LWLT should be assigned to the genus Streptomyces. On the basis of evidence from polyphasic study, strain TRM66268-LWLT is designated as representing a novel species of the genus Streptomyces, for which the name Streptomyces polyasparticus sp. nov. is proposed. The type strain is TRM66268-LWLT (CCTCC AA 2020003T = LMG32106T).

The leguminous trees Vachellia seyal (Del.) and Prosopis juliflora (Swartz) DC and their association with rhizobial strains from the root-influence zone of the grass Sporobolus robustus Kunth

This study focused on the genetic, symbiotic and phenotypic diversity of a collection of 12 Vachellia seyal and Prosopis juliflora rhizobial strains that were isolated from soils influenced by the roots of the grass Sporobolus robustus. Phylogenetic trees were constructed based on data from the 16S rRNA gene, from a multi-locus sequence analysis (MLSA) of the housekeeping genes recA, atpD, glnA and gyrB, as well as from the symbiotic genes nodA, nodC and nifH. Most of the mesorhizobial strains were clustered with the M. plurifarium type strain ORS 1032T and the 16S rRNA gene was as discriminatory as the other housekeeping genes. The Ensifer collection was closely related to E. alkalisoli YIC 4027T and E. fredii LMG 6217T using 16S rRNA and MLSA. Amplification of the Rhizobium collection was only observed with the 16S rRNA gene, where the strains were clustered with Rhizobium sp. 1B, Rhizobium sp. SEMIA 6411, Rhizobium sp. AS1-101a and Rhizobium sp. ORS 3441. The phylogeny of the nifH gene from Mesorhizobium and Ensifer strains were similar to those of nodulation genes (nodA and nodC). The nodA-nodC and nifH gene phylogenies were not consistent with those of 16S rRNA and the housekeeping genes. Ensifer genospecies tolerated high salinity better than the other genotypes after testing on a salt content gradient up to 49‰. We conclude that the rhizosphere of Sporobolus tussocks harbors a rhizobial microbiota, which could be capable of helping V. seyal and P. juliflora to grow better in saline soils.

Redesign of ultrasensitive and robust RecA gene circuit to sense DNA damage.

SummarySOS box of the recA promoter, PVRecA from Vibrio natriegens was characterized, cloned and expressed in a probiotic strain E. coli Nissle 1917. This promoter was then rationally engineered according to predicted interactions between LexA repressor and PVRecA. The redesigned PVRecA‐AT promoter showed a sensitive and robust response to DNA damage induced by UV and genotoxic compounds. Rational design of PVRecA coupled to an amplification gene circuit increased circuit output amplitude 4.3‐fold in response to a DNA damaging compound mitomycin C. A TetR‐based negative feedback loop was added to the PVRecA‐AT amplifier to achieve a robust SOS system, resistant to environmental fluctuations in parameters including pH, temperature, oxygen and nutrient conditions. We found that E. coli Nissle 1917 with optimized PVRecA‐AT adapted to UV exposure and increased SOS response 128‐fold over 40 h cultivation in turbidostat mini‐reactor. We also showed the potential of this PVRecA‐AT system as an optogenetic actuator, which can be controlled spatially through UV radiation. We demonstrated that the optimized SOS responding gene circuits were able to detect carcinogenic biomarker molecules with clinically relevant concentrations. The ultrasensitive SOS gene circuits in probiotic E. coli Nissle 1917 would be potentially useful for bacterial diagnosis.

Computational analysis of LexA regulons in Proteus species.

To gain a general understanding of the SOS system in Proteus species, in this study LexA-binding sites and the LexA regulons in 23 Proteus genomes were first predicted by phylogenetic footprinting server, then with Proteus vulgaris as an example, the expression of LexA regulon in iron limitation was investigated by proteomic analysis and quantitative reverse transcription polymerase chain reaction (RT-qPCR) method. The results showed that LexA proteins were highly conserved in Proteus species, and were in a close phylogenetic relationship with those in Gram-negative bacteria; the core SOS response genes lexA and recA were found in all the 23 genomes, indicating that this system was widely distributed in this genus; besides that, putative LexA-binding sites were also found in the upstream sequences of some genes involved in other biological processes such as biosynthesis, drug resistance, and stress response. Proteomic and RT-qPCR analyses showed that under iron deficient condition, the expression of lexA, recA and sulA was transcriptionally upregulated (p < 0.05), lexA was also translationally upregulated but recA was on the contrary (p < 0.05), whereas another SOS response gene dinI was transcriptionally downregulated (p < 0.01). These results indicated that in response to iron deficiency, the members of LexA regulon were not regulated by the same way, suggesting the existence of a precise regulation mechanism of SOS response in P. vulgaris. In conclusion, this study provided a preliminary understanding of the SOS system in Proteus species, which laid the foundation for further investigation of its roles in SOS response and other biological processes.

Studies of Phylogeny, Symbiotic Functioning and Ecological Traits of Indigenous Microsymbionts Nodulating Bambara Groundnut (Vigna subterranea L. Verdc)in Eswatini.

Rhizobial microsymbionts of grain legumes are ubiquitous in soils and exhibit a wide range of diversity with respect to colony morphology, genetic variability, biochemical characteristics, and phylogenetic relationships. This study assessed the phylogenetic positions of rhizobial microsymbionts of Bambara groundnut from Eswatini exhibiting variations in morpho-physiology, adaptive characteristics, and N2-fixing efficiency. Theisolates' ERIC-PCR profiles revealed the presence of high genetic variation among them. These test isolates also exhibited differences in pH tolerance and IAA production. Multilocus sequence analysis based on the 16S rRNA, atpD, glnII, gyrB, and recA gene sequences of representative test isolates closely aligned themto the type strains ofBradyrhizobium arachidis, B. manausense, B. guangdongense, B. elkanii, and B. pachyrhizi. However, some isolates showed a high divergence from the known reference type strains, indicating that they may represent species yet to be properly characterized and described. Functional characterization in the glasshouse revealed that most of the isolates from the contrasting Agro-ecologies of Eswatini were efficient in N2 fixation, and therefore elicited greater stomatal conductance and photosynthetic rates in the homologous Bambara groundnut. Of the 75 isolates tested, 51% were more effective than the commercial Bradyrhizobium sp.strain CB756, with relative symbiotic effectiveness ranging from 138 to 308%. The findings of this study indicated that the analysis ofhousekeeping genes and functional traits of Bambara-nodulating microsymbionts can provide a clear view for understanding and predicting rhizobial community structure across environmental gradients.

Phylogenetic Revision of the Genus Aliivibrio: Intra- and Inter-Species Variance Among Clusters Suggest a Wider Diversity of Species.

Genus Aliivibrio is known to harbor species exhibiting bioluminescence as well as pathogenic behavior affecting the fish farming industry. Current phylogenetic understanding of Aliivibrio has largely remained dormant after reclassification disentangled it from the Vibrio genus in 2007. There is growing evidence of wider diversity, but until now the lack of genomes and selective use of type strains have limited the ability to compare and classify strains firmly. In this study, a total of 143 bacterial strains, including 51 novel sequenced strains, were used to strengthen phylogenetic relationships in Aliivibrio by exploring intra-species and inter-species relations. Multilocus sequence analysis (MLSA), applying the six housekeeping genes 16S ribosomal RNA (rRNA), gapA, gyrB, pyrH, recA, and rpoA, inferred 12 clades and a singular branch in Aliivibrio. Along with four new phylogenetic clades, the MLSA resolved prior inconsistencies circumscribing Aliivibrio wodanis and formed a unique clade we propose as the novel species Aliivibrio sp. “friggae.” Furthermore, phylogenetic assessment of individual marker genes showed gyrB, pyrH, and recA superior to the 16S rRNA gene, resolving accurately for most species clades in Aliivibrio. In this study, we provide a robust phylogenetic groundwork for Aliivibrio as a reference point to classification of species.

Diversity of Root Nodule-Associated Bacteria of Diverse Legumes Along an Elevation Gradient in the Kunlun Mountains, China.

Bacteria in root nodules of legumes play important roles in promoting plant growth. In this study, we investigated root nodule-associated bacteria isolated from leguminous plants along an elevation gradient on the northern slope of the Kunlun Mountains, China, using a cultivation approach. In total, 300 isolates were obtained from seven legume species within six ecological zones. Isolates were identified based on 16S rRNA gene phylogenetic analysis and potential rhizobia were further identified using a recA gene phylogeny. Among the isolates, Bacillales (particularly Bacillus) were the dominant isolates from all host legumes and all elevations (63.5%), followed by Rhizobiales (13%) and Pseudomonadales (11.7%). Less than 3% of the isolates belonged to Burkholderiales, Paenibacillales, Enterobacteriales, Actinomycetales, Sphingomonadales, Xanthomonadales, Chitinophagales, Brevibacillales, Staphylococcales, or Mycobacteriales. A few elevation-specific patterns emerged within the Bacillales and Pseudomonadales. For example, isolates related to the psychrotroph Bacillus psychrosaccharolyticus were only isolated from the highest elevation sites (>3,500 m) whereas those related to the mesophile Bacillus endophyticus were only isolated from lowest elevation sites (1,350 m), suggestive of a role of soil temperature in their distribution. Similarly, isolates related to Pseudomonas brassicacearum were the dominant Pseudomonadales isolates, but they were only isolated from middle and low elevations (<3,200 m). A total of 39 isolates belonged to the Rhizobiales, 36 of which were confirmed to the genus level using the recA gene. In all, Rhizobiales isolates were obtained from five different host legumes spanning the entire elevation gradient. Those from the low-elevation Qira Desert-Oasis Transition Zone (1,350–1,960 m) suggested some patterns of host preference. For example, most isolates from Albizia julibrissin formed a monophyletic group related to Rhizobium lemnae and most from Alhagi sparsifolia were closely related to Ensifer kummerowiae. In general, this study shows that most bacteria associated with root nodules of legumes are widely distributed in distinct ecological zones within a single geographic region but suggests that both climate and host interactions may influence their distributions.

Diversity of rhizobia isolated from Tunisian arid soils capable of forming nitrogen-fixing symbiosis with Anthyllis henoniana

Soils in arid and infra-arid regions of Tunisia (where rainfall does not exceed 180 mm) suffer from rapid degradation of vegetation cover and loss of biodiversity. To limit the progress of desertification, it is necessary to promote the cultivation of autochthonous plants and particularly legumes, pioneer plants used for the restoration of poor soils. 50 strains were isolated from soils of five locations throughout arid areas in Tunisia using Anthyllis henoniana, a woody Saharan legume. 16S rRNA gene sequencing and multilocus phylogenetic analysis using recA and dnaK genes grouped rhizobial isolates into six clades, three of them related to the genus Rhizobium (36%), particularly R. leguminosarum, R. sullae and R. radiobacter, two related to Ensifer (56%), E. meliloti and E. numidicus, and one related to Neorhizobium (8%). Twelve selected strains were assayed for symbiotic efficiency using A. henoniana. Strains related to Ensifer and R. sullae were symbiotically efficient; demonstrating for the first time that they are natural symbionts of A. henoniana in Tunisian arid soils. Strains Ensifer sp. AZ2.5 and Rhizobium sp. AE4.7 were the most efficient in nitrogen fixation, suggesting that these strains could be interesting inoculants to enhance A. henoniana adaptation and growth in poorly vegetated regions.

Multilocus Sequence Typing of Leuconostoc mesenteroides Strains From the Qinghai-Tibet Plateau.

Leuconostoc mesenteroides strains were a type of epiphytic bacterium widely used in fermented foods and products in the biochemical and pharmaceutical industries but data on its presence in foods from Qinghai-Tibet Plateau in China was scarce. In this study, molecular analysis based on multilocus sequence typing (MLST) with eight housekeeping genes (pyrG, groeL, rpoB, recA, uvrC, murC, carB, and pheS) was carried out on 45 L. mesenteroides strains isolated from different plants and dairy products from Qinghai-Tibet Plateau in China. The objective of this study was to perform genetic diversity analysis and explore the relationship between strains and isolate samples or separate regions. A total of 25 sequence types (STs) were identified with a diversity of up to 55.6%, which were grouped into one clonal complexes (CCs), 3 doublets and 17 singletons by eBURST. The results of minimum spanning tree and clustering analysis indicated these L. mesenteroides strains from the Qinghai-Tibet Plateau were relatively weakly related to the isolated region. However, there was a close relationship between the genotypes of L. mesenteroides strains and the type of the isolated sample, which was consistent with the results of API 50CH. The MLST scheme presented in this study provides a shareable and comparable sequence database and enhances our knowledge of the population diversity of L. mesenteroides strains which will be further used for the selection of industrial strains.

Streptomyces scabichelini sp. nov., isolated from soil

A novel actinomycete, designated strain HC44T, was isolated from a soil sample collected from Hacibektaş, Turkey, and characterized using a polyphasic approach. The strain had morphological characteristics and chemotaxonomic properties identical to those of members of the genus Streptomyces. Phylogenetic analyses based on 16S rRNA gene sequence comparisons revealed that HC44T clustered with members of the genus Streptomyces and the highest 16S rRNA gene sequence similarity values were obtained with Streptomyces vastus NBRC 13094T (97.6 %) and Streptomyces kalpinensis TRM 46509T (96.9 %). Multi-locus sequence analysis (MLSA) based on five housekeeping genes (atpD, gyrB, recA, rpoB and trpB) showed that the MLSA evolutionary distance value was 0.043 between strain HC44T and S. vastus NBRC 13094T. Whole-cell hydrolysates contained ll-diaminopimelic acid, glucose, mannose and ribose. The predominant menaquinones were MK-9(H6) and MK-9(H8). The major polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol and phosphatidylinositol mannoside. The DNA G+C content of the draft genome sequence, consisting of 11.2 Mbp, was 69.8 mol%. On the basis of polyphasic taxonomic evidence, strain HC44T represents a novel species of the genus Streptomyces, for which the name Streptomyces scabichelini sp. nov. is proposed. The type strain is HC44T (=DSM 106874T=KCTC 39872T).

Core and Accessory Genome Analysis of Vibrio mimicus.

Vibrio mimicus is an emerging pathogen, mainly associated with contaminated seafood consumption. However, little is known about its evolution, biodiversity, and pathogenic potential. This study analyzes the pan-, core, and accessory genomes of nine V. mimicus strains. The core genome yielded 2424 genes in chromosome I (ChI) and 822 genes in chromosome II (ChII), with an accessory genome comprising an average of 10.9% of the whole genome for ChI and 29% for ChII. Core genome phylogenetic trees were obtained, and V. mimicus ATCC-33654 strain was the closest to the outgroup in both chromosomes. Additionally, a phylogenetic study of eight conserved genes (ftsZ, gapA, gyrB, topA, rpoA, recA, mreB, and pyrH), including Vibrio cholerae, Vibrio parilis, Vibrio metoecus, and Vibrio caribbenthicus, clearly showed clade differentiation. The main virulence genes found in ChI corresponded with type I secretion proteins, extracellular components, flagellar proteins, and potential regulators, while, in ChII, the main categories were type-I secretion proteins, chemotaxis proteins, and antibiotic resistance proteins. The accessory genome was characterized by the presence of mobile elements and toxin encoding genes in both chromosomes. Based on the genome atlas, it was possible to characterize differential regions between strains. The pan-genome of V. mimicus encompassed 3539 genes for ChI and 2355 genes for ChII. These results give us an insight into the virulence and gene content of V. mimicus, as well as constitute the first approach to its diversity.