Characterization of a novel root-associated diazotrophic rare PGPR taxa, Aquabacter pokkalii sp. nov., isolated from pokkali rice: new insights into the plant-associated lifestyle and brackish adaptation.

Salinity impacts crop growth and productivity and lowers the activities of rhizosphere microbiota. The identification and utilization of habitat-specific salinity-adapted plant growth-promoting rhizobacteria (PGPR) are considered alternative strategies to improve the growth and yields of crops in salinity-affected coastal agricultural fields. In this study, we characterize strain L1I39T, the first Aquabacter species with PGPR traits isolated from a salt-tolerant pokkali rice cultivated in brackish environments. L1I39T is positive for 1-aminocyclopropane-1-carboxylate deaminase activity and nitrogen fixation and can promote pokkali rice growth by supplying fixed nitrogen under a nitrogen-deficient seawater condition. Importantly, enhanced plant growth and efficient root colonization were evident in L1I39T-inoculated plants grown under 20% seawater but not in zero-seawater conditions, identifying brackish conditions as a key local environmental factor critical for L1I39T-pokkali rice symbiosis. Detailed physiological studies revealed that L1I39T is well-adapted to brackish environments. In-depth genome analysis of L1I39T identified multiple gene systems contributing to its plant-associated lifestyle and brackish adaptations. The 16S rRNA-based metagenomic study identified L1I39T as an important rare PGPR taxon. Based on the polyphasic taxonomy analysis, we established strain L1I39T as a novel Aquabacter species and proposed Aquabacter pokkalii sp nov. Overall, this study provides a better understanding of a marine-adapted PGPR strain L1I39T that may perform a substantial role in host growth and health in nitrogen-poor brackish environments.

Sphingomonas pokkalii sp. nov., a novel plant associated rhizobacterium isolated from a saline tolerant pokkali rice and its draft genome analysis.

Three strains L3B27T, 3CNBAF, L1A4 isolated from a brackish cultivated pokkali rice rhizosphere were characterised using a polyphasic taxonomic approach. Phylogenetic analysis based on 16S rRNA and recA gene sequences revealed that these strains were highly similar among each other and formed a separate monophyletic cluster within the genus Sphingomonas with Sphingomonas pituitosa DSM 13101T, Sphingomonas azotifigens DSM 18530T and Sphingomonas trueperi DSM 7225T as their closest relatives sharing 97.9-98.3% 16S rRNA similarity and 91.3-94.0% recA similarity values, respectively. The average nucleotide identity (ANI), average amino acid identity (AAI) and digital DNA-DNA hybridisation (dDDH) values between L3B27T (representative of the novel strains) and its phylogenetically closest Sphingomonas species were well below the established cut-off <94% (ANI/AAI) and <70% (dDDH) for species delineation. Further, the novel strains can be distinguished from its closest relatives based on several phenotypic traits. Thus, based on the polyphasic approach, we describe a novel Sphingomonas species for which the name Sphingomonas pokkalii sp. nov (type strain L3B27T=KCTC 42098T=MCC 3001T) is proposed. In addition, the novel strains were characterised for their plant associated properties and found to possess several phenotypic traits which probably explain its plant associated lifestyle. This was further confirmed by the presence of several plant associated gene features in the genome of L3B27T. Also, we could identify gene features which may likely involve in brackish water adaptation. Thus, this study provides first insights into the plant associated lifestyle, genome and taxonomy of a novel brackish adapted plant associated Sphingomonas.

Description of Vogesella oryzae sp. nov., isolated from the rhizosphere of saline tolerant pokkali rice.

Three strains, namely L3B39(T), L3D16, and L1E9, were obtained while studying the cultivable rhizosphere bacteria of saline tolerant pokkali rice, at Kerala, India. The novel strains were negative for many plant growth promoting plate assays such as phytohormone and siderophore production, 1-aminocyclopropane-1-carboxylate (ACC) deaminase and growth in nitrogen free agar medium but found to utilize malic acid, citrate, D-glucose, L-arabinose, and D-maltose, important components of the plant root exudates, indicating that they are normal plant rhizosphere residents without yet known benefits to the plant. The 16S rRNA gene analysis placed these strains in the genus Vogesella, forming a separate branch independent of the previously described type strains of this genus in all tree making algorithms applied. Vogesella perlucida DS-28(T) was the type strain with highest 16S rRNA sequence similarity (97.59%). DNA-DNA hybridization values among these novel strains were above 85% andthat with Vogesella perlucida LMG 24214(T) was below 50%. Phenotypically, the novel strains can be differentiated from Vogesella perlucida LMG 24214(T) by many characters such as NaCl tolerance, growth temperature, and utilization of L-arabinose, D-maltose, and citrate. These novel strains contain C16:1ω6c/C16:1ω7c and C16:0 as major fatty acids, ubiquinone Q-8 as the major respiratory quinone, and phosphatidylethanolamine and phosphatidylglycerol as major polar lipids. Based on the results obtained from the polyphasic taxonomic approach we conclude that the strains belong to a novel Vogesella species for which the name Vogesella oryzae sp.nov. is proposed. The type strain is L3B39(T) (= LMG 28272(T)=DSM 28780(T)).

Vibrio mangrovi sp. nov., a diazotrophic bacterium isolated from mangrove-associated wild rice (Poteresia coarctata Tateoka).

The taxonomic status of a nitrogen-fixing bacterium, strain MSSRF38(T), isolated from the rhizosphere of mangrove-associated wild rice (Porteresia coarctata Tateoka), in Pichavaram, India, was studied using a polyphasic approach. Phylogenetic analyses based on 16S rRNA gene sequences indicated that the novel strain MSSRF38(T) was most closely related to Vibrio ruber DSM 16370(T) (98.3% gene sequence similarity), Vibrio rhizosphaerae DSM 18581(T) (98.2% sequence similarity) and <96% to the remaining Vibrio species. Multilocus sequence analysis using ftsZ, gapA, gyrB and mreB genes showed low levels of gene sequence similarities (82-90%) with all species of the genus Vibrio with validly published names, indicating that strain MSSRF38(T) occupies a distinct phylogenetic position. DNA-DNA hybridization experiments showed that strain MSSRF38(T) had <70% DNA-DNA similarity to its closest neighbours V. ruber DSM 16370(T) (27.4%) and V. rhizosphaerae DSM 18581(T) (12.1%). Strain MSSRF38(T) could be differentiated from its relatives on the basis of several phenotypic characteristics. The major fatty acids were feature 3 (including C(16:1omega)7c and/or C(15:0) iso 2-OH), C16:0, C(18:1omega)7c, C(14:0) and C(12:0). The DNA G+C content was 45.4 mol%. Based on genotypic, phenotypic, chemotaxonomic and DNA-DNA analyses, the name Vibrio mangrovi sp. nov. (type strain MSSRF38(T)=LMG 24290(T)=DSM 19641(T)) is proposed for this novel taxon.