Sphingomonas sp. Hbc-6 alters Arabidopsis metabolites to improve plant growth and drought resistance by manipulating the microbiome.

Drought significantly affects crop productivity and poses a considerable threat to agricultural ecosystems. Plant growth-promoting bacteria (PGPB) and plant microbiome play important roles in improving drought resistance and plant performance. However, the response of the rhizosphere microbiota to PGPB during the development of plants and the interaction between inoculum, microbiota, and plants under drought stress remain to be explored. In the present study, we used culturomic, microbiomic, and metabonomic analyses to uncover the mechanisms by which Sphingomonas sp. Hbc-6, a PGPB, promotes Arabidopsis growth and enhances drought resistance. We found that the rhizosphere microbiome assembly was interactively influenced by developmental stage, Hbc-6, and drought; the bacterial composition exhibited three patterns of shifts with developmental stage: resilience, increase, and decrease. Drought diminished microbial diversity and richness, whereas Hbc-6 increased microbial diversity and helped plants recruit specific beneficial bacterial taxa at each developmental stage, particularly during the bolting stage. Some microorganisms enriched by Hbc-6 had the potential to promote carbon and nitrogen cycling processes, and 86.79 % of the isolated strains exhibited PGP characteristics (for example Pseudomonas sp. TA9). They jointly regulated plant physiological metabolism (i.e., upregulated drought resistant-facilitating substances and reduced harmful substances), thereby stimulating the growth of Arabidopsis and increasing plant biomass under drought stress conditions. Collectively, these results indicate that Hbc-6 mediates plant growth and drought resistance by affecting the microbiome. The study thus provides novel insights and strain resources for drought-resistant, high-yielding crop cultivation and breeding.

Production of Algal Biomass and High-Value Compounds Mediated by Interaction of Microalgal Oocystis sp. KNUA044 and Bacterium Sphingomonas KNU100.

There is growing interest in the production of microalgae-based, high-value by-products as an emerging green biotechnology. However, a cultivation platform for Oocystis sp. has yet to be established. We therefore examined the effects of bacterial culture additions on the growth and production of valuable compounds of the microalgal strain Oocystis sp. KNUA044, isolated from a locally adapted region in Korea. The strain grew only in the presence of a clear supernatant of Sphingomonas sp. KNU100 culture solution and generated 28.57 mg/l/d of biomass productivity. Protein content (43.9 wt%) was approximately two-fold higher than carbohydrate content (29.4 wt%) and lipid content (13.9 wt%). Oocystis sp. KNUA044 produced the monosaccharide fucose (33 µg/mg and 0.94 mg/l/d), reported here for the first time. Fatty acid profiling showed high accumulation (over 60%) of polyunsaturated fatty acids (PUFAs) compared to saturated (29.4%) and monounsaturated fatty acids (9.9%) under the same culture conditions. Of these PUFAs, the algal strain produced the highest concentration of linolenic acid (C18:3 ω3; 40.2%) in the omega-3 family and generated eicosapentaenoic acid (C20:5 ω3; 6.0%), also known as EPA. Based on these results, we suggest that the application of Sphingomonas sp. KNU100 for strain-dependent cultivation of Oocystis sp. KNUA044 holds future promise as a bioprocess capable of increasing algal biomass and high-value bioactive by-products, including fucose and PUFAs such as linolenic acid and EPA.

Purification and characterization of a novel medium-chain ribitol dehydrogenase from a lichen-associated bacterium Sphingomonas sp.

Sugar alcohols (polyols) are abundant carbohydrates in lichen-forming algae and transported to other lichen symbionts, fungi, and bacteria. Particularly, ribitol is an abundant polyol in the lichen Cetraria sp. Polyols have important physiological roles in lichen symbiosis, but polyol utilization in lichen-associated bacteria has been largely unreported. Herein, we purified and characterized a novel ribitol dehydrogenase (RDH) from a Cetraria sp.-associated bacterium Sphingomonas sp. PAMC 26621 grown on a minimal medium containing D-ribitol (the RDH hereafter referred to as SpRDH). SpRDH is present as a trimer in its native form, and the molecular weight of SpRDH was estimated to be 39 kDa by SDS-PAGE and 117 kDa by gel filtration chromatography. SpRDH converted D-ribitol to D-ribulose using NAD+ as a cofactor. As far as we know, SpRDH is the first RDH belonging to the medium-chain dehydrogenase/reductase family. Multiple sequence alignments indicated that the catalytic amino acid residues of SpRDH consist of Cys37, His65, Glu66, and Glu157, whereas those of short-chain RDHs consist of Ser, Tyr, and Lys. Furthermore, unlike other short-chain RDHs, SpRDH did not require divalent metal ions for its catalytic activity. Despite SpRDH originating from a psychrophilic Arctic bacterium, Sphingomonas sp., it had maximum activity at 60°C and exhibited high thermal stability within the 4-50°C range. Further studies on the structure/function relationship and catalytic mechanism of SpRDH will expand our understanding of its role in lichen symbiosis.

Effects of Intranasal Pseudorabies Virus AH02LA Infection on Microbial Community and Immune Status in the Ileum and Colon of Piglets.

Pseudorabies virus (PRV) variants broke out in china since 2011, causing high fever, respiratory distress, systemic neurological symptoms, and diarrhea in piglets. This study investigated the effect of intranasal PRV variant (AH02LA) infection on ileal and colonic bacterial communities and immune status in piglets. Ten piglets (free of PRV) were assigned to PRV variant and control groups (uninfected). At day 5 after inoculation, all piglets were euthanized. No PRV was detected in the ileal and colonic mucosa. In the PRV group, we observed up-regulation of specific cytokines gene expression, down-regulation of intestinal barrier-related gene expression, and reduction of secretory immunoglobulin A (sIgA) concentration in the ileum and colon. PRV infection increased the diversity of ileal bacterial community composition. PRV infection reduced the abundance of some beneficial bacteria (Lactobacillus species in the ileum and colon; butyrate-producing bacteria species in the colon) and increased the abundance of potentially pathogenic Fusobacterium nucleatum in the ileum and Sphingomonas paucimobilis in the colon. Moreover, PRV infection decreased concentrations of the beneficial lactate in the ileum and butyrate in the colon. However, this study does not allow to evaluate whether the observed changes are directly due to the PRV infection or rather to indirect effects (fever, clinical signs and changes in diet), and will be our next research content. In summary, our findings provide evidence that intranasal PRV infection directly or indirectly brings gut health risks and implications, although no PRV was detected in the ileum and colon.

Growth, Viability, and Death of Planktonic and Biofilm Sphingomonas desiccabilis in Simulated Martian Brines.

Aqueous solutions on Mars are theorized to contain very different ion compositions than those on Earth. To determine the effect of such solutions on typical environmental micro-organisms, which could be released from robotic spacecraft or human exploration activity, we investigated the resistance of Sphingomonas desiccabilis to brines that simulate the composition of martian aqueous environments. S. desiccabilis is a desiccation-resistant, biofilm-forming microbe found in desert crusts. The viability of cells in both planktonic and biofilm forms was measured after exposure to simulated martian brines. Planktonic cells showed a loss of viability over the course of several hours in almost all of the seven brines tested. Biofilms conferred greater resistance to all the brines, including those with low water activity and pH, but even cells in biofilms showed a complete loss of viability in <6 h in the harsher brines and in <2 days in the less harsh brines. One brine, however, allowed the microbes to maintain viability over several days, despite having a water activity and pH lower and ionic strength higher than brines that reduced viability over the same timescales, suggesting important ion-specific effects. These data show that biofilm-forming cells have a greater capacity to resist martian aqueous extremes, but that evaporative or deliquescent brines are likely to be destructive to many organisms over relatively short timescales, with implications for the habitability of Mars and for micro-organisms dispersed by robotic or human explorers.

Rizobacterias promotoras de crecimiento en Guarianthe skinneri (Orchidaceae) / Growth-promoting rhizobacteria from Guarianthe skinneri (Orchidaceae)

Resumen La orquídea Guarianthe skinneri está incluida en la norma NOM-059-ECOL-2010 de México como una especie amenazada. Con el fin de estudiar las BPCV (bacterias promotoras del crecimiento vegetal) en esta orquídea, se recolectaron 10 raíces de diferentes plantas para aislar bacterias asociadas a las raíces, que se analizaron mediante pruebas in vitro como: producción de AIA, fijación de nitrógeno, interacción con el hongo micorrízico Thanatephorus sp. cepa RG26 y solubilización de fosfato. De los 71 aislados bacterianos se caracterizaron 10 cepas mediante secuenciación con el marcador 16s rADN y se identificaron seis cepas: Sphingomonas sp., Sinorhizobium sp., Bacillus sp., Nocardia cerradoensis, Bacillus megaterium y Burkholderia phytofirmans. Se observó que la bacteria Sinorhizobium sp. produjo mayor cantidad de AIA (69.189 µg/ml) y Bacillus sp. presentó mayor reducción de acetileno (10.251 nmol cultivo/96 h). En las interacciones de las bacterias y el hongo RG26 se presentaron cuatro categorías (sumamente positivo, positivo, antagonismo 50-50 e inhibición). En relación a la solubilización de fosfato, la bacteria Burkholderia phytofirmans presentó mayor IS a las 48 y 96 hr con IS de 3.11 y 3.48, respectivamente. Los resultados indican que Bacillus sp. pudiera tener las mejores características para promover el desarrollo de la orquídea G. skinneri mediante la inoculación de semillas y plántulas.

Abstract The Guarianthe skinneri orchid is included in NOM-059-ECOL-2010, Mexico standard as an endangered species. In order to study PGPR (promoting growth plant rhizobacteria) from this orchid, 10 roots were collected from different plants to isolate bacteria associated with the roots, which were analyzed by in vitro tests such as: production of AIA, nitrogen fixation, interaction with the mycorrhizal fungus Thanatephorus sp. strain RG26 and phosphate solubilization. We obtain 71 bacterial isolates, 10 strains of them were characterized by sequencing with the 16d rDNA marker identifying six bacteria: Sphingomonas sp. Sinorhizobium sp. Bacillus sp. Nocardia cerradoensis, Bacillus megaterium and Burkholderia phytofirmans. We observed that the bacterium Sinorhizobium sp. produced a greater amount of AIA (69.189 μg/ml) and Bacillus sp. performed greater acetylene reduction (10.251 nmol cultivo/96h). In the interactions of the bacteria and the fungus RG26, four categories were presented (extremely positive, positive, antagonism 50-50 and inhibition). In relation to the solubilization of phosphate, Burkholderia phytofirmans presented higher IS after 48 and 96 hr with an IS of 3.11 and 3.48, respectively. The results indicate that Bacillus sp. it could have the best characteristics to promote the development of the G. skinneri orchid by inoculating seeds and seedlings. Rev. Biol. Trop. 66(3): 953-968. Epub 2018 September 01.

Stereoselective accumulations of hexachlorocyclohexanes (HCHs) are correlated with Sphingomonas spp. in agricultural soils across China.

The wide usage of hexachlorocyclohexanes (HCHs) as pesticides has caused soil pollution and adverse health effects through direct contact or bioaccumulation in the food chain. This study quantified major HCH isomers in farmland topsoils across China, and evaluated their correlations with microbial community structure, function, and abiotic variables (e.g., moisture, pH, and temperature). Recalcitrant ß-HCH was more abundant than α-, γ-, and δ-HCHs, and α-HCH enantiomeric fractions (EF) were larger than 0.5, indicating preferential degradation of (-)-α-HCH. Sphingomonas was not only a predominant population (especially in samples collected in the south), but also a promising biomarker indicating total- and ß-HCH residuals, and EF values of α-HCH. Soil moisture and temperature were among the most influential factors that structured the diversity and function of soil microbial communities. The results suggested that increasing soil moisture (in the range of 5-45%) would benefit the growth of HCH-degrading populations and the enrichment of HCH-degradation related pathways. Revealing the site-specific relationships between topsoil physical, chemical, and microbial properties will benefit the in situ bioremediation of farmlands with relatively low HCH residuals across the world.

Differential regulation of phenanthrene biodegradation process by kaolinite and quartz and the underlying mechanism.

Natural and cost-effective materials such as minerals can serve as supportive matrices to enhance biodegradation of polycyclic aromatic hydrocarbons (PAHs). In this study we evaluated and compared the regulatory role of two common soil minerals, i.e. kaolinite and quartz in phenanthrene (a model PAH) degradation by a PAH degrader Sphingomonas sp. GY2B and investigated the underlying mechanism. Overall kaolinite was more effective than quartz in promoting phenanthrene degradation and bacterial growth. And it was revealed that a more intimate association was established between GY2B and kaolinite. Si and O atoms on mineral surface were demonstrated to be involved in GY2B-mineral interaction. There was an higher polysaccharide/lipid content in the EPS (extracellular polymeric substances) secreted by GY2B on kaolinite than on quartz. Altogether, these results showed that differential bacterial growth, enzymatic activity, EPS composition as well as the interface interaction may explain the effects minerals have on PAH biodegradation. It was implicated that different interface interaction between different minerals and bacteria can affect microbial behavior, which ultimately results in different biodegradation efficiency.

Inoculation of soil with an Isoproturon degrading microbial community reduced the pool of "real non-extractable" Isoproturon residues.

During pesticides degradation, biogenic non-extractable residues ("apparent NER") may not share the same environmental fate and risks with the "real NER" that are bound to soil matrix. It is not clear how microbial community (MC) inoculation for pesticides degradation would influence the NER composition. To investigate degradation efficiency of pesticides Isoproturon (IPU) and NER composition following MC inoculation, clay particles harboring MC that contains the IPU degrading strain, Sphingomonas sp., were inoculated into soil receiving 14C-labeled IPU addition. Mineralization of IPU was greatly enhanced with MC inoculation that averagely 55.9% of the applied 14C-IPU was consumed up into 14CO2 during 46 days soil incubation. Isoproturon degradation was more thorough with MC than that in the control: much less amount of metabolic products (4.6% of applied IPU) and NER (35.4%) formed in MC treatment, while the percentages were respectively 30.3% for metabolites and 49.8% for NER in the control. Composition of NER shifted with MC inoculation, that relatively larger amount of IPU was incorporated into the biogenic "apparent NER" in comparison with "real NER". Besides its well-recognized role on enhancing mineralization, MC inoculation with clay particles benefits soil pesticides remediation in term of reducing "real NER" formation, which has been previously underestimated.

Insights into the genome and proteome of Sphingomonas paucimobilis strain 20006FA involved in the regulation of polycyclic aromatic hydrocarbon degradation.

In order to study the mechanisms regulating the phenanthrene degradation pathway and the intermediate-metabolite accumulation in strain S. paucimobilis 20006FA, we sequenced the genome and compared the genome-based predictions to experimental proteomic analyses. Physiological studies indicated that the degradation involved the salicylate and protocatechuate pathways, reaching 56.3% after 15 days. Furthermore, the strain degraded other polycyclic aromatic hydrocarbons (PAH) such as anthracene (13.1%), dibenzothiophene (76.3%), and fluoranthene. The intermediate metabolite 1-hydroxy-2-naphthoic acid (HNA) accumulated during phenanthrene catabolism and inhibited both bacterial growth and phenanthrene degradation, but exogenous-HNA addition did not affect further degradation. Genomic analysis predicted 126 putative genes encoding enzymes for all the steps of phenanthrene degradation, which loci could also participate in the metabolism of other PAH. Proteomic analysis identified enzymes involved in 19 of the 23 steps needed for the transformation of phenanthrene to trichloroacetic-acid intermediates that were upregulated in phenanthrene cultures relative to the levels in glucose cultures. Moreover, the protein-induction pattern was temporal, varying between 24 and 96 h during phenanthrene degradation, with most catabolic proteins being overexpressed at 96 h-e. g., the biphenyl dioxygenase and a multispecies (2Fe-2S)-binding protein. These results provided the first clues about regulation of expression of phenanthrene degradative enzymes in strain 20006FA and enabled an elucidation of the metabolic pathway utilized by the bacterium. To our knowledge the present work represents the first investigation of genomic, proteomic, and physiological studies of a PAH-degrading Sphingomonas strain.