•Straw input and rice-growing stimulate soil nitrite oxidizing potential (NO).•Soil NO is related to a shift in Nitrospira-like NOB community structure.•Nitrospira community shift was significantly affected by pH, NH4+ and moisture.•Nitrobacter-like NOB was just not significantly affected in this system.•Nitrospira are more sensitive to straw input and rice-growing than Nitrobacter.Nitrite oxidation is recognized as an essential process of biogeochemical nitrogen cycling in agricultural ecosystems. How nitrite-oxidizing bacteria (NOB) respond to land managements (the effect from the long-term straw incorporation and environmental variability caused by the shift from the upland stage to the paddy stage) in a rapeseed-rice rotation field remains unclear. We found the nitrite oxidation (NO) in soils increased from the upland stage to the paddy stage. An inhibitory effect of the long-term straw incorporation on NO was detectable in the upland stage. The abundance of Nitrospira was always greater than Nitrobacter, and it was affected by the rice-growing and straw incorporation while Nitrobacter was not. NO correlated positively with the abundance of Nitrospira and with soluble sulfate (SO42 −), soil moisture, pH and NH4+. The high-throughput sequencing analysis of the nitrite oxidoreductase nxrA and nxrB genes for Nitrobacter- and Nitrospira-like NOB was performed respectively. The dominating (relative abundance > 1%) operational taxonomic units (OTUs) from Nitrobacter were closely related to Nitrobacter hamburgensis, whereas those from Nitrospira were affiliated with or related to lineage II, lineage V and several unknown groups. Heatmap analysis showed that a few dominant Nitrobacter OTUs were affected by the straw treatment or the rice-growing, and half of the dominant Nitrospira ones were explained by at least one of the variables. Multi-response permutation procedure (MRPP) and redundancy analyses showed that the Nitrospira-like NOB community changes were significantly shaped by the land managements and the soil chemical properties, including pH, moisture and NH4+, whereas that of the Nitrobacter-like NOB community was not. These results suggested that Nitrospira are more sensitive than Nitrobacter to land management in acid and fertilized soils of a rapeseed-rice rotation field trial.Download high-res image (268KB)Download full-size image

The global regulatory molecule (p)ppGpp is synthesized under limited nutrition conditions and involves in many cellular processes in bacteria. (p)ppGpp has been reported to affect biofilm formation in several bacterial species. Here, we found that deletion of (p)ppGpp synthase genes of Pseudomonas putida KT2440 led to enhanced biofilm formation in polystyrene microtitre plates. Besides, the pellicle of this mutant formed at the air-liquid interface lost the robust structure and became frail. The biofilm formation and its structure are mainly determined by exopolysaccharides (EPSs) and adhesins. Transcriptional analysis of four EPS operons designated as pea, peb, alg and bcs and two adhesin genes nominated as lapA and lapF showed that the deletion of (p)ppGpp synthase genes increased the expression of peb, bcs and lapA but repressed the expression of pea and lapF. Furthermore, expression of the regulation factor FleQ was significantly augmented in (p)ppGpp-synthase mutants while the expression of sigma factor RpoS was reduced. Since FleQ and RpoS play important roles in regulating expression of EPS and adhesin genes, (p)ppGpp may mediate the synthesis of biofilm matrix via influencing these regulators to control the biofilm formation and pellicle structure.

Adsorption and desorption of salmon sperm DNA on bacteria (Bacillus thuringiensis, Pseudomonas putida), two different colloidal fractions (organic and inorganic clay) from an Alfisol, minerals (montmorillonite, kaolinite and goethite) and colloid–bacteria composites were studied. Similar adsorption capacity and affinity of DNA were observed on two bacterial cells. However, the two bacterial strains played different roles in affecting the adsorption of DNA on the composites of soil colloidal particles with bacteria. The introduction of B. thuringiensis in soil colloids and minerals systems dramatically promoted DNA adsorption on colloidal particles especially organic clay, while P. putida decreased the adsorption of DNA on kaolinite and goethite. Electrostatic force and ligand exchange are regarded to be the major driving forces involved in the adsorption of DNA on bacterial cells, montmorillonite, soil colloids and goethite. Presence of bacteria enhanced the proportion of DNA adsorption on soil colloidal particles by electrostatic force and depressed that by ligand exchange process. Information obtained in this study is of fundamental significance for the understanding of the ultimate fate of extracellular DNA in soil systems.

In this study, Fourier transform infrared spectroscopy, potentiometric titration, along with sorption experiments using chemically modified bacteria were conducted to compare the behavior of Gram-positive Bacillus thuringiensis and Gram-negative Escherichia coli as sorbents for Cu2+ and Cd2+ ions. The IR spectra showed that there were obvious changes connected with the–COOH groups for Cu(II)- and Cd(II)-loaded B. thuringiensis. A three site non-electrostatic model provides an excellent fit to the titration curves of both E. coli and B. thuringiensis with the first corresponding pKa values of 4.16 ± 0.18 and 3.30 ± 0.24, respectively, implying that B. thuringiensis contains more carboxyl groups than E. coli. Chemical modification and metal sorption experiments further confirmed that carboxyl groups may play a more important role in the binding Cd(II) and Cu(II) for B. thuringiensis than for E. coli, which could be attributed to the higher concentration of carboxyl sites of B. thuringiensis than E. coli. These results show that the main chemical functional group responsible for the binding of metal ions on B. thuringiensis and E. coli is obviously different because of the different composition and structure of bacterial cell walls. The concentrations of the chemical functional groups on bacterial cell walls are regarded to govern their role in the binding of metal ions and affect the affinity between the bacterial cells and metal ions.

The filamentous cyanobacterium Anabaena sp. PCC 7120 forms nitrogen-fixing heterocysts after deprivation of combined nitrogen. Under such conditions, vegetative cells provide heterocysts with photosynthate and receive fixed nitrogen from the latter. Heterocyst envelope contains a glycolipid layer and a polysaccharide layer to restrict the diffusion of oxygen into heterocysts. Low-Molecular-Weight protein tyrosine phosphatases (LMW-PTPs) are involved in the biosynthesis of exopolysaccharides in bacteria. Alr5068, a protein from Anabaena sp. PCC 7120, shows significant sequence similarity with LMW-PTPs. In this study we characterized the enzymatic properties of Alr5068 and showed that it can dephosphorylate several autophosphorylated tyrosine kinases (Alr2856, Alr3059 and All4432) of Anabaena sp. PCC 7120 in vitro. Several conserved residues among LMW-PTPs are shown to be essential for the phosphatase activity of Alr5068. Overexpression of alr5068 results in a strain unable to survive under diazotrophic conditions, with the formation of morphologically mature heterocysts detached from the filaments. Overexpression of an alr5068 allele that lost phosphatase activity led to the formation of heterocyst with an impaired polysaccharide layer. The alr5068 gene was upregulated after nitrogen step-down and its mutation affected the expression of hepA and hepC, two genes necessary for the formation of the heterocyst envelope polysaccharide (HEP) layer. Our results suggest that Alr5068 is associated with the production of HEP in Anabaena sp. PCC 7120.

S-(3, 4-dichlorobenzyl) isothiourea (A22) has been reported to specifically inhibit the function of MreB, an actin-like protein in rod-shaped bacteria. This study investigated the role of A22 in cyanobacterium Anabaena sp. strain PCC 7120, which can form nitrogen-fixing heterocysts under combined-nitrogen deprivation. Results indicated that A22 could inhibit cell growth, cause abnormal cellular morphology and bring about asymmetric cell division and irregular DNA distribution. However, A22 has little effect on heterocyst formation. An A22-resistant mutant named C23 was isolated by growing cells on A22-containing plates. It had normal appearance of cell shape, division and DNA content when treated by A22. However, this mutant retained a wild-type allele of mreB.

•High efficiency of growing fungus strain XLA in reducing Cr6+.•Biotransformation, biosorption and bioaccumulation are involved in reducing Cr6+.•Extra/intracellular defense systems of XLA are contributed to Cr6+ detoxification.•Responses of Cr defense systems to various Cr6+ levels are diverse.In the study, the capability of Paecilomyces lilacinus XLA (CCTCC: M2012135) to reduce Cr6+ and its main antagonistic mechanisms to Cr6+ were experimentally evaluated. Activated growing fungus XLA efficiently reduced over 90% Cr6+ in the media with Cr6+ concentration below 100 mg L−1 at pH 6 after 14 days. After 1-day exposure to 100 mg L−1 Cr6+, nearly 50% of Cr6+ was reduced. Moreover, SO42− stimulated Cr6+ reduction, whereas other interferential ions inhibited Cr6+ reduction. The interaction mechanisms between XLA and Cr6+ mainly involve biotransformation, biosorption, and bioaccumulation, as detected by electron microscopy and chemical methods. The lower concentrations of Cr6+ (5 and 50 mg L−1) stimulated the activities of superoxide dismutase (SOD), catalase (CAT) and glutathione (GSH) level in XLA, respectively, but the higher concentration of Cr6+ (150 mg L−1) decreased the enzymatic activities and GSH concentration. The results implied that SOD, CAT and GSH were defensive guards to the oxidant stress produced by Cr6+. All these extracellular/intracellular defense systems endowed XLA with the ability to resist and detoxify Cr6+ by transforming its valent species. The fungus XLA could efficiently reduce Cr6+ under different environmental conditions (pH, interferential ions, and concentration). Moreover, XLA could endure the high concentration of Cr6+ probably due to its high biotransformation capability of Cr6+ and intracellular antioxidant systems for the detoxification of ROS generated by external Cr6+. All these results suggested that the fungus XLA can be applied to remediation of Cr6+-contaminated environments.Download high-res image (182KB)Download full-size image

The cyanobacterium Anabaena (Nostoc) sp. PCC 7120 is a model for cyanobacterial cell differentiation studies. pDU1, an endogenous plasmid in Nostoc sp. PCC 7524, is used as the only cyanobacterial replicon for Anabaena (Nostoc) studies. However, the relative copy numbers of pDU1-based plasmids in Anabaena (Nostoc) sp. PCC 7120 are not well studied. We found that the relative plasmid copy number of one such vector, pRL25T, varied widely, especially when the vector carried a recombinant insert, under different conditions, ranging from 0.53 to 1812 per chromosome in different recombinant strains tested, either in independent clones of the same strain or in the same clone under different growth conditions. The phenotypes caused by pRL25T-driven expression of green fluorescent protein or the GAF domain of Pkn41 or Pkn42 varied depending on the independent clones analyzed. This phenotypic variation correlated with the relative plasmid copy number present in cells.