Comparative physiological and transcriptomic analysis of two salt-tolerant soybean germplasms response to low phosphorus stress: role of phosphorus uptake and antioxidant capacity.

BACKGROUND: Phosphorus (P) and salt stress are common abiotic stressors that limit crop growth and development, but the response mechanism of soybean to low phosphorus (LP) and salt (S) combined stress remains unclear. RESULTS: In this study, two soybean germplasms with similar salt tolerance but contrasting P-efficiency, A74 (salt-tolerant and P-efficient) and A6 (salt-tolerant and P-inefficient), were selected as materials. By combining physiochemical and transcriptional analysis, we aimed to elucidate the mechanism by which soybean maintains high P-efficiency under salt stress. In total, 14,075 differentially expressed genes were identified through pairwise comparison. PageMan analysis subsequently revealed several significantly enriched categories in the LP vs. control (CK) or low phosphorus + salt (LPS) vs. S comparative combination when compared to A6, in the case of A74. These categories included genes involved in mitochondrial electron transport, secondary metabolism, stress, misc, transcription factors and transport. Additionally, weighted correlation network analysis identified two modules that were highly correlated with acid phosphatase and antioxidant enzyme activity. Citrate synthase gene (CS), acyl-coenzyme A oxidase4 gene (ACX), cytokinin dehydrogenase 7 gene (CKXs), and two-component response regulator ARR2 gene (ARR2) were identified as the most central hub genes in these two modules. CONCLUSION: In summary, we have pinpointed the gene categories responsible for the LP response variations between the two salt-tolerant germplasms, which are mainly related to antioxidant, and P uptake process. Further, the discovery of the hub genes layed the foundation for further exploration of the molecular mechanism of salt-tolerant and P-efficient in soybean.

A nuclease-toxin and immunity system for kin discrimination in Myxococcus xanthus.

The use of toxin to attack neighbours and immunity proteins to protect against toxin has been observed in bacterial conflicts, including kin discrimination. Here, we report a novel nuclease-toxin and its immunity protein function in the colony-merger incompatibility, a kind of bacterial kin discrimination, in Myxococcus xanthus DK1622. The MXAN_0049 gene was determined to be a genetic determinant for colony-merger incompatibility, and the incompatibility could be eliminated by deletion of the upstream co-transcribed MXAN_0050 gene. We demonstrated that the MXAN_0050 protein was a nuclease, and MXAN_0049 protein was able to bind to MXAN_0050 to block nuclease activity in vitro. Expression of MXAN_0050 in Escherichia coli inhibited cellular growth, and the inhibition effect could be recovered by co-expression of MXAN_0049. We found that deletion of the PAAR-encoding gene (MXAN_0044) or the type VI secretion system led to the colony-merger and co-existence with the ΔMXAN_0049 mutant, suggesting that they were associated with colony-merger incompatibility. Homologues of the nuclease-toxin and cognate immunity pair are widely distributed in bacteria. We propose a simplified model to explain the kin discrimination mechanism mediated by the nuclease-toxin and immunity protein.© 2018 Society for Applied Microbiology and John Wiley & Sons Ltd.

[Fabrication and Surface-Enhanced Raman Scattering Research on Polystyrene Nanospheres Arrays].

To research surface-enhanced Raman scattering effect of different diameters' polystyrene nanospheres (Ag-coated) substrates, we had prepared a new surface-enhanced Raman scattering substrate. It was prepared by using spin-coating and self-assembled to form monolayer and hexagonal close-packed (HCP) order colloidal-crystal films with PS nanospheres of different diameters. Then, a 30 nm Ag film was deposited on the PS arrays with magnetron sputtering. We detected Raman scattered spectrum by means of Roman spectroscopy instrument with Rhodamine R6G as a probe molecule, and the conclusion was drawn that all the substrates of Ag-coated PS nanospheres displayed stronger Raman enhancement. With the increase of the diameter of the microspheres, the degree of fluctuation of polystyrene nanospheres array was continuously strengthened (increasing roughness), and the SERS signal was enhanced gradually. The signal peak reached a maximum when the ball diameter was 600 nm, and then we aquired an optimal SERS substrate. Simultaneously, we found that the SERS spectra of R6G of a high signal to noise ratio was obtained on the substrate, and associated with the benzene ring within a range of CC double bond stretching vibration characteristic spectrum and the inner surface and the outer surface deformation vibration characteristic spectra were significantly enhanced. The single and large-scale Raman scattering substrate presents especial nanostructure of high and low distribution. The depth and the gap between different PS microspheres show the obvious differences, which can significantly improve the surface morphology structure of Ag film and Raman scattering effect of PS nanospheres substrate. The substrate presenting special arrays' structure has significantly great potential in exploring molecule of chemical and biological filed.

[Surface-enhanced raman spectra of thymine calculated by DFT and ab inition HF].

In the present paper, B3LYP (Becke's three-parameter hybrid method with the Lee, Yang, and Parr gradient corrected correlation functional) and HF (Hartree-Fock) methods at 6-31+G* * (C, H, N, O)/LANL2DZ(Ag) level were used to optimize molecular configurations of thymine. Base on the optimized structure, the normal Raman spectrum (NRS) of thymine and the surface-enhanced Raman spectrum (SERS) of thymine adsorbed on Ag and Ag2 were calculated, which were compared with the experimental values. The calculation results indicated that the result of the DFT for NRS was more approximate to the ever reported experimental date than those of HF results. A better conformity of SERS was found between the HF computed and the experimental results. At the end, detailed analysis of the Raman spectrum and more comprehensive assignments of the vibration mode for thymine were studied by the software of GaussView.

Competitive Interactions Between Incompatible Mutants of the Social Bacterium Myxococcus xanthus DK1622.

Due to the high similarity in their requirements for space and food, close bacterial relatives may be each other's strongest competitors. Close bacterial relatives often form visible boundaries to separate their swarming colonies, a phenomenon termed colony-merger incompatibility. While bacterial species are known to have many incompatible strains, it is largely unclear which traits lead to multiple incompatibilities and the interactions between multiple incompatible siblings. To investigate the competitive interactions of closely related incompatible strains, we mutated Myxococcus xanthus DK1622, a predatory bacterium with complex social behavior. From 3392 random transposon mutations, we obtained 11 self-identification (SI) deficient mutants that formed unmerged colony boundaries with the ancestral strain. The mutations were at nine loci with unknown functions and formed nine independent SI mutants. Compared with their ancestral strain, most of the SI mutants showed reduced growth, swarming and development abilities, but some remained unchanged from their monocultures. When pairwise mixed with their ancestral strain for co-cultivation, these mutants exhibited improved, reduced or unchanged competitive abilities compared with the ancestral strain. The sporulation efficiencies were affected by the DK1622 partner, ranging from almost complete inhibition to 360% stimulation. The differences in competitive growth between the SI mutants and DK1622 were highly correlated with the differences in their sporulation efficiencies. However, the competitive efficiencies of the mutants in mixture were inconsistent with their growth or sporulation abilities in monocultures. We propose that the colony-merger incompatibility in M. xanthus is associated with multiple independent genetic loci, and the incompatible strains hold competitive interaction abilities, which probably determine the complex relationships between multiple incompatible M. xanthus strains and their co-existence strategies.

Myxobacterial community is a predominant and highly diverse bacterial group in soil niches.

Although many molecular ecological surveys have been conducted, there is little concerning the details of specific bacterial groups, resulting in an incomplete understanding of the microorganismal composition and community structures in the environment. Myxobacteria are micropredators that are metabolically active in the soil microbial food web and have typically been considered minority components of soil bacterial communities. In this study, we surveyed the percentage of myxobacteria in a single soil sample via pyrosequencing on combined universal libraries of the V3-V4 and V6-V8 hypervariable regions of the 16S rRNA gene. Surprisingly, myxobacteria accounted for 4.10% of the bacterial community and 7.5% of the total operational taxonomic units at the 3% similarity level in the soil, containing almost all of the cultivated myxobacterial families or genera. To testify the appearance of myxobacteria in soil niches, we retrieved myxobacteria-related 16S rRNA gene sequences of 103 high-throughput sequencing data sets obtained from public databases. The results indicated that myxobacteria-related sequences were among the predominant groups in these data sets accounting for 0.4-4.5% of bacterial communities. The abundance of myxobacterial communities were correlated with site temperature, carbon-to-nitrogen ratio and pH values. Based on these results, we discussed the survival strategies of myxobacterial community in soil.

The existence and diversity of myxobacteria in lake mud - a previously unexplored myxobacteria habitat.

Myxobacteria are widely distributed in soil and oceanic sediment with a phylogeographic separation at high levels of classification. However, it is unclear whether freshwater environments, from which there has been no isolation report of myxobacteria since 1981, are habitats for myxobacteria. In this study, we investigated the presence of myxobacteria in lake mud using a two-step strategy. First, we constructed two universal bacterial libraries from the V3-V4 (V34) and V6-V8 (V678) hypervariable regions of 16S rRNA gene sequences. High-throughput 454 pyrosequencing revealed that myxobacteria were one of the major bacterial groups in the lake mud. They accounted for 5.77% of the total sequences and 7.52% of the total operational taxonomic units (OTUs) at a phylogenetic distance of 0.03. The community composition and taxonomic structure of the mud myxobacterial community were further analysed using myxobacteria-enriched libraries targeting the V34 and V678 regions, which were amplified with Cystobacterineae- and Sorangineae-specific primer pairs respectively. Phylogenetic analysis showed that the limnetic myxobacteria exhibited closer relationships to their soil than their marine relatives, but there were also exclusive taxa of limnetic myxobacteria detected. These results, together with a survey on available GenBank data, indicate that lake mud is a primary habitat for myxobacteria.

Phylogeographic separation of marine and soil myxobacteria at high levels of classification.

Microorganisms are globally dispersed and are able to proliferate in any habitat that supports their lifestyles, which, however, has not yet been explored in any specific microbial taxon. The social myxobacteria are considered typical soil bacteria because they have been identified in various terrestrial samples, a few in coastal areas, but none in other oceanic environments. To explore the prevalence of marine myxobacteria and to investigate their phylogenetic relationships with their terrestrial counterparts, we established myxobacteria-enriched libraries of 16S rRNA gene sequences from four deep-sea sediments collected at depths from 853 to 4675 m and a hydrothermal vent at a depth of 204 m. In all, 68 different myxobacteria-related sequences were identified from randomly sequenced clones of the libraries of different samples. These myxobacterial sequences were diverse but phylogenetically similar at different locations and depths. However, they were separated from terrestrial myxobacteria at high levels of classification. This discovery indicates that the marine myxobacteria are phylogeographically separated from their terrestrial relatives, likely because of geographic separation and environment selection.