Performance evaluation of the effect of humic acid on Anammox granular sludge: Apparent morphology, nitrogen removal and microbial community.

Humic acid (HA) is a typical refractory organic matter, so it is of great significance to investigate its effect on the performance of Anammox granular sludge. When the dosage of HA ≤ 50 mg/L, HA promotes the total nitrogen removal rate (NRR) to 1.45 kg/(m3·day). When HA was between 50 and 100 mg/L, the NRR of Anammox was stable. At this time, the adsorption of HA causes the sludge to gradually turn from red to brown, but the activities of heme and enzymes showed that its capacity was not affected. When HA levels reached 250 mg/L, the NRR dropped to 0.11 kg/(m3·day). Moderate HA levels promoted the release of extracellular polymeric substance (EPS), but excessive HA levels lead to a decrease in EPS concentrations. HA inhibited Anammox activity, which indirectly hindered the transmission of substrate and accumulated substrate toxicity. Although HA promoted the increase of heterotrophic microbial abundance in Anammox system, the microbial diversity decreased gradually. With the increase of HA concentration, the abundance of Candidatus_Brocadia, the main functional microorganism of Anammox system, decreased gradually, while the abundance of Candidatus_Kuenenia increased gradually.

Plant Disease Resistance-Related Pathways Recruit Beneficial Bacteria by Remodeling Root Exudates upon Bacillus cereus AR156 Treatment.

The environmentally friendly biological control strategy that relies on beneficial bacterial inoculants to improve plant disease resistance is a promising strategy. Previously, it has been demonstrated that biocontrol bacteria treatments can change the plant rhizosphere microbiota but whether plant signaling pathways, especially those related to disease resistance, mediate the changes in rhizosphere microbiota has not been explored. Here, we investigated the complex interplay among biocontrol strains, plant disease resistance-related pathways, root exudates, rhizosphere microorganisms, and pathogens to further clarify the biocontrol mechanism of biocontrol bacteria by using plant signaling pathway mutants. Bacillus cereus AR156, which was previously isolated from forest soil by our laboratory, can significantly control tomato bacterial wilt disease in greenhouse and field experiments. Moreover, compared with the control treatment, the B. cereus AR156 treatment had a significant effect on the soil microbiome and recruited 35 genera of bacteria to enrich the rhizosphere of tomato. Among them, the relative rhizosphere abundance of nine genera, including Ammoniphilus, Bacillus, Bosea, Candidimonas, Flexivirga, Brevundimonas, Bordetella, Dyella, and Candidatus_Berkiella, was regulated by plant disease resistance-related signaling pathways and B. cereus AR156. Linear correlation analysis showed that the relative abundances of six genera in the rhizosphere were significantly negatively correlated with pathogen colonization in roots. These rhizosphere bacteria were affected by plant root exudates that are regulated by signaling pathways. IMPORTANCE Our data suggest that B. cereus AR156 can promote the enrichment of beneficial microorganisms in the plant rhizosphere by regulating salicylic acid (SA) and jasmonic acid (JA)/ethylene (ET) signaling pathways in plants, thereby playing a role in controlling bacterial wilt disease. Meanwhile, Spearman correlation analysis showed that the relative abundances of these beneficial bacteria were correlated with the secretion of root exudates. Our study reveals a new mechanism for SA and JA/ET signals to participate in the adjustment of plant resistance whereby the signaling pathways adjust the rhizosphere microecology by changing the root exudates and thus change plant resistance. On the other hand, biocontrol strains can utilize this mechanism to recruit beneficial bacteria by activating disease resistance-related signaling pathways to confine the infection and spread of pathogens. Finally, our data also provide a new idea for the in-depth study of biocontrol mechanisms.

Effects of salinity on nitrite and elemental sulfur accumulation in a double short-cut sulfur autotrophic denitrification process.

Double short-cut sulfur autotrophic denitrification (DSSADN) coupled with Anammox is of great significance in the low-carbon treatment of nitrogen-containing wastewater. In order to achieve high salinity autotrophic nitrogen removal, the effects of different salinities on the accumulation characteristics of NO2--N and S0 and microorganisms in DSSADN process were studied. The results showed that the effect of salinity on the DSSADN process could be categorized into the stimulation, stable, and inhibition. When the salinity gradually increased to 2.5 %, the highest NO2--N production rate (NiPR) and S0 production rate (S0PR) of DSSADN were 0.54 kg/(m3·d) and 1.1 kg/(m3·d) respectively. NiPR and S0PR gradually decreased as the salinity increased to more than 3 %. However, salinity had a relatively low impact on nitrite accumulation efficiency and S0 accumulation efficiency, which were 80 % and 81.5 %, respectively, when the salinity reached 5 %. Salinity has a great influence on the structure and abundance of microbial communities in the system.

Mitogen-Activated Protein Kinases Associated Sites of Tobacco Repression of Shoot Growth Regulates Its Localization in Plant Cells.

Plant defense and growth rely on multiple transcriptional factors (TFs). Repression of shoot growth (RSG) is a TF belonging to a bZIP family in tobacco, known to be involved in plant gibberellin feedback regulation by inducing the expression of key genes. The tobacco calcium-dependent protein kinase CDPK1 was reported to interact with RSG and manipulate its intracellular localization by phosphorylating Ser-114 of RSG previously. Here, we identified tobacco mitogen-activated protein kinase 3 (NtMPK3) as an RSG-interacting protein kinase. Moreover, the mutation of the predicted MAPK-associated phosphorylation site of RSG (Thr-30, Ser-74, and Thr-135) significantly altered the intracellular localization of the NtMPK3-RSG interaction complex. Nuclear transport of RSG and its amino acid mutants (T30A and S74A) were observed after being treated with plant defense elicitor peptide flg22 within 5 min, and the two mutated RSG swiftly re-localized in tobacco cytoplasm within 30 min. In addition, triple-point mutation of RSG (T30A/S74A/T135A) mimics constant unphosphorylated status, and is predominantly localized in tobacco cytoplasm. RSG (T30A/S74A/T135A) showed no re-localization effect under the treatments of flg22, B. cereus AR156, or GA3, and over-expression of this mutant in tobacco resulted in lower expression levels of downstream gene GA20ox1. Our results suggest that MAPK-associated phosphorylation sites of RSG regulate its localization in tobacco, and that constant unphosphorylation of RSG in Thr-30, Ser-74, and Thr-135 keeps RSG predominantly localized in cytoplasm.

[Establishment of a mouse model of cyclophosphamide-induced thrombocytopenia and determination of platelet function in this model].

OBJECTIVE: To establish a mouse model of acute hematopoietic failure and explore the pathological basis of platelet changes in bone marrow suppression. METHODS: An initial large dose of cyclophosphamide (200 mg/kg) was injected through the tail veins of the mice, follow by daily intraperitoneal injection starting on the next day for 7 d. The number and morphology of the cells in the peripheral blood and bone marrow were observed by means of cell counting and smear, respectively, with the platelet aggregation determined using ADP. The coagulation time was measure by turbidimetry. RESULTS: The amount of platelets, erythrocytes, leukocytes and the nucleated cells in mice bone marrow was significantly lowered in the mouse models in comparison with the control mice, and the normal hematopoietic tissues were depressed. The blood platelets in the models were lowered by 49%, an amplitude of declination significantly greater than that occurring in the erythrocytes and leukocytes (by 28% and 25% respectively). Though the mean platelet volume and percent platelet aggregation underwent no obvious changes, the coagulation time was significantly shortened. CONCLUSION: The methods we described for establishing mouse models of hematopoietic failure induced by cyclophosphamide is rapid and highly efficient, and may facilitate ready preparation of mouse models of thrombocytopenia which respond strongly to cyclophosphamide without blood platelets function impairment.