Up-regulation of growth-related gene expression in tobacco by volatile compounds released by Bacillus velezensis WSW007.

In order to investigate the mechanism of plant growth promoting (PGP) effects of strain Bacillus velezensis WSW007, its PGP traits and production of volatile organic compounds (VOCs) were tested. The effects of VOCs produced by strain WSW007 on plant growth were observed by co-culturing this strain with tobacco seedlings in I-plates. Meanwhile, the effects of VOCs on tobacco gene expression were analysed by a transcriptome analysis and VOCs were identified by solid phase micro extraction coupled with gas chromatography-mass spectrometry (SPME-GC-MS) analysis. As results, strains WSW007 produced acetic acid and siderophore, and could solubilize phosphate; while it also significantly increased the fresh weight of tobacco seedlings via production of VOCs. In transcriptome analysis, plants co-cultured with strain WSW007 presented the highest up-regulated expression for the genes involved in plant growth and development processes, implying that the bacterial VOCs played a role as regulator of plant gene expression. Conclusively, the up-regulation in expression of growth- and development-related genes via VOCs production is an important PGP mechanism in strain B. velezensis WSW007.

Soluble humic acid suppresses plant immunity and ethylene to promote soybean nodulation.

Symbiotic nitrogen fixation (SNF) is a crucial process for nitrogen geochemical cycling and plant-microbe interactions. Water-soluble humic acid (WSHM), an active component of soil humus, has been shown to promote SNF in the legume-rhizobial symbiosis, but its molecular mechanism remains largely unknown. To reveal the SNF-promoting mechanism, we conducted transcriptomic analysis on soybean treated with WSHM. Our findings revealed that up- and downregulated differentially expressed genes (DEGs) were mainly involved in plant cell-wall/membrane formation and plant defence/immunity in the early stage, while the late stage was marked by the flavonoid synthesis and ethylene biosynthetic process. Further study on representative DEGs showed that WSHM could inhibit GmBAK1d-mediated immunity and BR signalling, thereby promoting rhizobial colonisation, infection, and nodulation, while not favoring pathogenic bacteria colonisation on the host plant. Additionally, we also found that the ethylene pathway is necessary for promoting the soybean nodulation by WSHM. This study not only provides a significant advance in our understanding of the molecular mechanism of WSHM in promoting SNF, but also provides evidence of the beneficial interactions among the biostimulator, host plant, and soil microbes, which have not been previously reported.

Chelativorans petroleitrophicus sp. nov., a paraffin oil-degrading bacterium isolated from a mixture of oil-based drill cuttings and paddy soil.

We isolated a paraffin oil-degrading bacterial strain from a mixture of oil-based drill cutting and paddy soil, and characterized the strain using a polyphasic approach. The Gram-positive, aerobic, rod-shaped and non-spore-forming strain (SCAU 2101T) grew optimally at 50 °C, pH 7.0 and 0.5 % (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequence indicated that the strain represented a distinct clade in the genus Chelativorans, neighbouring Chelativorans intermedius LMG 28482T (97.1 %). The genome size and DNA G+C content of the strain were 3 969 430 bp and 63.1 mol%, respectively. Whole genome based phylogenomic analyses showed that the average nucleotide identity and digital DNA-DNA hybridization values between strain SCAU 2101T and C. intermedius LMG 28482T were 77.5 and 21.2 %, respectively. The major respiratory quinone was Q-10. The dominant fatty acids were C19 : 0 cyclo ω8c (50.6 %), summed feature 8 (C18 : 1 ω7c and/or C18 : 1 ω6c; 22.5 %) and C18 : 0 (13.8 %). The polar lipids of the strain included phosphatidylethanolamine, phosphatidylmonomethylethanolamine, phosphatidylglycerol, phosphatidylcholine and diphosphatidylglycerol. Based on the results, strain SCAU 2101T was considered to represent a novel species in the genus Chelativorans, for which the name Chelativorans petroleitrophicus sp. nov. is proposed. The type strain is SCAU 2101T (= CCTCC AB 2021125T=KCTC 92067T).

Volatile Organic Compounds of Streptomyces sp. TOR3209 Stimulated Tobacco Growth by Up-Regulating the Expression of Genes Related to Plant Growth and Development.

To investigate the mechanism underlying the plant growth-promoting (PGP) effects of strain Streptomyces sp. TOR3209, PGP traits responsible for indoleacetic acid production, siderophore production, and phosphate solubilization were tested by culturing the strain TOR3209 in the corresponding media. The effects of volatile organic compounds (VOCs) produced by the strain TOR3209 on plant growth were observed by co-culturing this strain with tobacco seedlings in I-plates. Meanwhile, the effects of VOCs on tobacco gene expression were estimated by performing a transcriptome analysis, and VOCs were identified by the solid-phase micro-extraction (SPME) method. The results showed positive reactions for the three tested PGP traits in the culture of strain TOR3209, while the tobacco seedlings co-cultured with strain TOR3209 revealed an increase in the fresh weight by up to 100% when compared to that of the control plants, demonstrating that the production VOCs was also a PGP trait. In transcriptome analysis, plants co-cultured with strain TOR3209 presented the highest up-regulated expression of the genes involved in plant growth and development processes, implying that the bacterial VOCs played a role as a regulator of plant gene expression. Among the VOCs produced by the strain TOR3209, two antifungal molecules, 2,4-bis(1,1-dimethylethyl)-phenol and hexanedioic acid dibutyl ester, were found as the main compounds. Conclusively, up-regulation in the expression of growth- and development-related genes via VOCs production is an important PGP mechanism in strain TOR3209. Further efforts to explore the effective VOCs and investigate the effects of the two main VOCs in the future are recommended.

Rhizobium chutanense sp. nov., isolated from root nodules of Phaseolus vulgaris in China.

Two Gram-stain-negative, rod-shaped bacterial strains (C5T and C16), isolated from root nodules of Phaseolus vulgaris L. in Jiangxi Province, PR China, were characterized by using a polyphasic taxonomical approach. The phylogenetic analysis of the 16S rRNA gene and three concatenated housekeeping genes (recA-glnII-atpD) revealed that C5T and C16 were members of the genus Rhizobium, yet were distinct from known species. The case for strain C5T representing a novel species was supported by genomic results. Pairwise digital DNA-DNA hybridization and average nucleotide identity values were much lower than the proposed and generally accepted species boundaries. The genome-based phylogenetic tree reconstructed by using the up-to-date bacterial core gene set consisting of 92 genes showed that the strains formed a monophyletic branch, further supporting this result. The symbiotic genes of nodC and nifH were identified in both strains; each could nodulate Phaseolus vulgaris and Glycine max but not Leucaena leucocephala, Pisum sativum or Medicago sativa plants. Major cellular fatty acids of C5T were summed feature 8 (C18 : 1 ω7c/C18 : 1 ω6c; 58.8 %), C18 : 1 ω7c 11-methyl (14.2 %) and C18 : 0 (8.1 %). The DNA G+C content of C5T was 61.4 mol%. Based on these genomic, chemotaxonomic and phenotypic characteristics, we propose a novel species: Rhizobium chutanense sp. nov. The type strain is C5T (=CCTCC AB 2018143T=LMG 30777T).

Rhizobium changzhiense sp. nov., isolated from effective nodules of Vicia sativa L. in North China.

Three fast-growing rhizobial strains isolated from effective nodules of common vetch (Vicia sativa L.) were characterized using a polyphasic approach. All three strains were assigned to the genus Rhizobium on the basis of the results of 16S rRNA gene sequence analysis. Phylogenetic analysis based on concatenated atpD-recA genes separated the strains into a distinct lineage represented by WYCCWR 11279T, which showed average nucleotide identity values of 95.40 and 93.61 % with the most similar phylogenetic type strains of Rhizobium sophorae CCBAU 03386T and Rhizobium laguerreae FB TT, respectively. The digital DNA-DNA hybridization relatedness values between WYCCWR 11279T and the closest related type strains were less than 70 %. Therefore, a novel rhizobial species is proposed, Rhizobium changzhiense sp. nov., and strain WYCCWR 11279T (=HAMBI 3709T=LMG 31534T) is designated as the type strain for the novel species.

Novel Butane-Oxidizing Bacteria and Diversity of bmoX Genes in Puguang Gas Field.

To investigate the diversity of butane-oxidizing bacteria in soils contaminated by long-term light hydrocarbon microseepage and the influence of butane on the soil microbial community, a quantitative study and identification of butane-oxidizing bacteria (BOB) in soils at the Puguang gas field were performed by DNA-based stable isotope probing (DNA-SIP). For the first time, two phylotypes corresponding to the genera Giesbergeria and Ramlibacter were identified as being directly involved in butane oxidation, in addition to the well-known light hydrocarbon degrader Pseudomonas. Furthermore, bmoX genes were strongly labeled by 13C-butane, and their abundances in gas field soils increased by 43.14-, 17.39-, 21.74-, and 30.14-fold when incubated with butane for 6, 9, 12, and 14 days, respectively, indicating that these bmoX-harboring bacteria could use butane as the sole carbon and energy source and they play an important role in butane degradation. We also found that the addition of butane rapidly shaped the bacterial community and reduced the diversity of bmoX genes in the gas field soils. These findings improve our understanding of BOB in the gas field environment and reveal the potential for their applications in petroleum exploration and bioremediation.

Syntrophic Interactions Within a Butane-Oxidizing Bacterial Consortium Isolated from Puguang Gas Field in China.

Butane oxidation by the hydrocarbon degradation bacteria has long been described, but little is known about the microbial interaction in this process. To investigate this interaction, the efficiency of butane oxidation was estimated in monocultures and co-cultures of six strains of butane-oxidizing bacteria (BOB) and a butanol-oxidizing strain. Results showed that the butane degradation velocity was at least 26 times higher in the co-culture of the seven strains (228.50 nmol h(-1)) than in the six individual monocultures (8.71 nmol h(-1)). Gas chromatographic analysis of metabolites in the cultures revealed the accumulation of butanol in the monocultures of BOB strains but not in the co-culture with the butanol-oxidizing strain. These results evidenced a novel syntrophic association between BOB and butanol-oxidizing bacteria in the butane oxidation. The BOB strains oxidized butane into butanol, but this activity was inhibited by the accumulated butanol in monocultures, whereas the removal of butanol by the butanol-oxidizing strain in co-culture could eliminate the suppression and improve the butane degradation efficiency. In the co-culture, both BOB and butanol-oxidizing bacteria could grow and the time needed for butane complete removal was shortened from more than 192 h to less than 4 h. The unsuppressed effect of the co-culture was also consistent with the results of reverse transcription quantitative real-time PCR (RT-qPCR) of bmoX gene because increased expression of this gene was detected during the syntrophic growth compared with that in monoculture, pointing to the upregulation of bmoX in the syntrophic interaction.

Novosphingobium tardum sp. nov., isolated from sediment of a freshwater lake.

A Gram-negative, non-motive, aerobic and non-spore-forming strain 16-28-2(T) isolated from freshwater sediment of Taihu Lake was characterized by using a polyphasic approach. The optimum growth conditions were found to be as follows: 28 °C, pH 6.5 and 0-0.5 % NaCl in YG liquid medium. The major fatty acids were identified to be summed feature 3 (consisting of C16:1 ω7c and/or C16:1 ω6c), summed feature 8 (consisting of C18:1 ω7c and/or C18:1 ω6c), C14:0 2-OH, C17:1 ω6c, C16:0 and C18:1 ω7c 11-methyl (>5 %). Strain 16-28-2(T) was found to contain diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine and sphingoglycolipid as the major polar lipids; and ubiquinone 10 (Q-10) as the major respiratory quinone. DNA G+C content of strain 16-28-2(T) was 63.5 mol % (Tm). A phylogenetic study of 16S rRNA gene indicated that strain 16-28-2(T) is a member of the genus Novosphingobium, with the highest 16S rRNA gene sequence similarity of 96.3 % with Novosphingobium lentum MT1(T) and below 96 % with the other Novosphingobium species. On the basis of the phylogenetic, phenotypic analyses and biochemical characterization, we suggest that strain 16-28-2(T) is a novel species in the genus Novosphingobium, for which the name Novosphingobium tardum sp. nov. is proposed. The type strain of N. tardum is 16-28-2(T) (=CGMCC 1.12989(T) =NBRC 110956(T)).

Cadmium(II) removal by a hyperaccumulator fungus Phoma sp. F2 isolated from blende soil.

A cadmium(II)-resistant fungus, strain F2, isolated from blende soil was identified as Phoma sp. by morphological study and internal transcribed spacer sequencing. This strain could accumulate 280 mg of Cd(II)/g dry weight mycelium. In liquid medium containing 163.8 mg Cd(II)/L, 96% of Cd(II) was removed by the actively growing mycelium. In addition, both oven-dried and lyophilized mycelium could effectively adsorb Cd(II). There were removed 91% and 46.2% of Cd(II) from 51.6 mg Cd(II)/L solution by lyophilized biomass and oven-dried biomass respectively. Transmission electron microscopy and energy-dispersive X-ray analysis showed the accumulation of Cd(II) in the mycelium cell walls. Our results demonstrated that Phoma sp. F2 was a hyperaccumulator for the removal of Cd(II) from contaminated soil and water.

The novel alkali tolerance function of tfxG in Sinorhizobium meliloti.

TfxG, one of the tfxABCDEFG cluster genes that code for trifolitoxin (TFX) production, was initially described in Rhizobium leguminosarum bv. trifolii T24. Although several genes in the tfx family have functions related to TFX production or resistance to TFX, the function of tfxG is largely unknown. Using cDNA-amplified fragment length polymorphism (cDNA-AFLP) analysis, we found that expression of the tfxG gene dramatically increased under alkaline culture conditions in Sinorhizobium meliloti CCBAU 81024. This result was confirmed by northern blot analysis. Mutagenesis of tfxG significantly decreased the viability of Sinorhizobium meliloti CCBAU 81024 under alkali stress. Complementation of the tfxG mutant strain using the functional tfxG gene recovered its alkali tolerance to a wild-type level. Genomic analysis of the tfxG gene suggests that choline and homoserine kinase domains may contribute to its alkali tolerance function. This is the first clear evidence that tfxG plays a crucial role in the alkali tolerance of S. meliloti CCBAU 81024, and the finding provides its biological function.