Limoniibacter endophyticus gen. nov., sp. nov., an alphaproteobacterium isolated from the roots of Limonium otolepis.

A Gram-negative bacterium, designated as strain YIM 690229T, was isolated from the roots of Limonium otolepis. The strain was able to grow at 10-40 °C (optimum, 28-37 °C), pH 6.0-8.0 (optimum, 7.0) and in the presence of up to 7% NaCl (w/v) (optimum, up to 2.5%). Comparative 16S rRNA gene sequence analysis revealed that strain YIM 690229T shared less than 93.9% sequence similarities with members within the order Rhizobiales, and was remotely related to members of the family Hyphomicrobiaceae. Strain YIM 690229T was characterized by the presence of Q-10 as the predominant respiratory lipoquinone. The major fatty acids (> 10%) detected were C18:1 ω7c, C16:0, anteiso-C15:0 and summed feature 4 (iso-C17:1 I and/or anteiso-C17:1 B). The polar lipids consisted of diphosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylmethylethanoamine and two unidentified lipids. The genomic DNA G + C content was 57.2 mol%. Data from this polyphasic taxonomy study suggested that strain YIM 690229T should be classified as a new species of a new genus within the family Hyphomicrobiaceae for which the name Limoniibacter endophyticus gen. nov., sp. nov., is proposed. The type species of the genus Limoniibacter gen. nov. is Limoniibacter endophyticus. The type strain of the species Limoniibacter endophyticus sp. nov. is YIM 690229T (= KCTC 42097T = JCM 30141T = CCTCC AB 2014130T = CGMCC 1.12906T).

Global transcriptome analysis of Mesorhizobium alhagi CCNWXJ12-2 under salt stress.

BACKGROUND: Mesorhizobium alhagi CCNWXJ12-2 is a α-proteobacterium which could be able to fix nitrogen in the nodules formed with Alhagi sparsifolia in northwest of China. Desiccation and high salinity are the two major environmental problems faced by M. alhagi CCNWXJ12-2. In order to identify genes involved in salt-stress adaption, a global transcriptional analysis of M. alhagi CCNWXJ12-2 growing under salt-free and high salt conditions was carried out. The next generation sequencing technology, RNA-Seq, was used to obtain the transcription profiles. RESULTS: We have compared the transcriptome of M. alhagi growing in TY medium under high salt conditions (0.4 M NaCl) with salt free conditions as a control. A total of 1,849 differentially expressed genes (fold change ≧ 2) were identified and 933 genes were downregulated while 916 genes were upregulated under high salt condition. Except for the upregulation of some genes proven to be involved in salt resistance, we found that the expression levels of protein secretion systems were changed under high salt condition and the expression levels of some heat shock proteins were reduced by salt stress. Notably, a gene encoding YadA domain-containing protein (yadA), a gene encoding trimethylamine methyltransferase (mttB) and a gene encoding formate--tetrahydrofolate ligase (fhs) were highly upregulated. Growth analysis of the three gene knockout mutants under salt stress demonstrated that yadA was involved in salt resistance while the other two were not. CONCLUSIONS: To our knowledge, this is the first report about transcriptome analysis of a rhizobia using RNA-Seq to elucidate the salt resistance mechanism. Our results showed the complex mechanism of bacterial adaption to salt stress and it was a systematic work for bacteria to cope with the high salinity environmental problems. Therefore, these results could be helpful for further investigation of the bacterial salt resistance mechanism.

Bacterial Community Structure and Potential Microbial Coexistence Mechanism Associated with Three Halophytes Adapting to the Extremely Hypersaline Environment.

Halophytes play a crucial ecological role in drought and saline-alkali environments. However, there is limited knowledge about the structure of bacterial communities and the potential microbial coexistence mechanism associated with halophytes. This study investigated the diversity and community structure of endophytic and rhizospheric bacteria associated with three halophytes by applying high-throughput sequencing and geochemistry analyses on the studied soils. We collected 18 plant and 21 soil samples, and sequenced the V3 and V4 hypervariable regions of the 16S rRNA gene using next-generation sequencing (NGS). We also assessed geochemistry of the studied soils. The research suggested that rhizospheric bacterial richness and diversity associated with three halophytes were all significantly higher than for endophytic bacteria. The microbial community analysis indicated that Actinobacteria, Firmicutes, Bacteroidetes and Proteobacteria were the dominating bacterial phyla. Most unassigned operational taxonomic units (OTUs) implied that the microbes associated with halophytes contained abundant potential novel taxa, which are significant microbial resources. The high-abundance OTU phylogenetic tree supported the above views as well. Additionally, network analysis indicated that some conditional rare taxa (CRT) also might be keystone taxa during halophyte microbial community construction. The results of non-metric multidimensional scaling (NMDS) ordination analysis indicated significant dissimilarities in the microbial community among different sample groups. Sixty-two biomarkers were detected from seven different sample groups by linear discriminant analysis effect size (LEFSe) analysis. Microbial functions predicted based on phylogenetic investigation of communities by reconstruction of unobserved states (PICRUSt2) demonstrated that the abundances of nitrogen metabolism genes of endophytic bacteria were significantly higher than in rhizobacteria. Environmental factor analysis confirmed that different soil properties have different degrees of influence on the abundance and composition of the microbiota. To better adapt to the extreme hypersaline environment, halophytes could specifically recruit some plant beneficial bacterial taxa, such as nitrogen-fixing bacteria and extremely halophilic or halotolerant bacteria, to help them robustly grow and proliferate. All our preliminary results highlight microbial diversity and community related to halophytes grown on saline-alkali land of arid areas. Simultaneously, this work also advanced our further understanding of the halophyte microbiome associated with plants, and their role in plant adaptation to the extremely hypersaline environment.