[Effects of Foliar Application of Silicon Fertilizers on Phyllosphere Bacterial Community and Functional Genes of Paddy Irrigated with Reclaimed Water].

Agricultural utilization of reclaimed water is considered to be an effective way to solve water shortage and reduce water environmental pollution. Silicon fertilizer can improve crop yield and quality and enhance crop resistance. The effect of foliar spray with silicon fertilizer on phyllosphere microbial communities remains lacking. In this study, a pot experiment was conducted to explore the effects of different types of silicon fertilizer on the composition and diversity of a phyllosphere bacterial community and the abundances of related functional genes in rice irrigated with reclaimed water. The results showed that Firmicutes, Proteobacteria, Actinobacteriota, Bacteroidota, and Verrucomicrobiota dominated the phyllosphere bacteria of rice. The relative abundance of Bacillus was higher than that of other treatments in RIS3. Reclaimed water irrigation significantly increased the relative abundances of the potential pathogens Pantoea and Enterobacter. The unclassified bacteria were also an important part of the bacterial community in the rice phyllosphere. Bacillus, Exiguobacterium, Aeromonas, and Citrobacter were significantly enriched by silicon fertilizer treatments. Functional prediction analysis showed that indicator species were mainly involved in metabolism and degradation functions, and the predicted functional groups of phyllosphere bacteria were attributed to chemoheterotrophy, aerobic chemoheterotrophy, nitrate reduction, and fermentation. Quantitative PCR results showed that AOA, AOB, and nifH genes were at low abundance levels in all treatments, and nirK genes was not significantly different among treatments. These results contribute to the in-depth understanding of the effects of foliar spray silicon fertilizer on the bacterial community structure and diversity of rice phyllosphere and provide a theoretical basis for the application of silicon fertilizer in reclaimed water irrigation agriculture.

[Effects of Soil Amendments on the Bacterial Diversity and Abundances of Pathogens and Antibiotic Resistance Genes in Rhizosphere Soil Under Drip Irrigation with Reclaimed Water].

Due to reclaimed water, irrigation can cause human health and environmental risks. Soil amendments are applied to reveal the abundance of pathogens and antibiotic resistance genes in rhizosphere soil irrigated by reclaimed water and to better understand the effects of environmental factors on the rhizosphere soil bacterial composition, which has guiding significance for the reasonable use of soil amendments. In this study, the effects of biochar, bioorganic fertilizer, humic acid, loosening soil essence, and corn vinasse on bacterial community diversity and certain gene abundances in rhizosphere soil under drip irrigation with reclaimed water were studied using high-throughput assays and quantitative PCR. The results showed that biochar significantly increased pH, organic matter, and total nitrogen contents in the rhizosphere soil. The corn vinasse significantly decreased soil pH and increased the contents of total nitrogen and total phosphorus but significantly increased the soil EC value (P<0.05). The effects of the five soil amendments on the α-diversity of rhizosphere bacteria were not significantly different. The bacterial community structure and diversity of rhizosphere bacteria were similar at different taxonomic levels, but their relative abundance was different. α-Proteobacteria, γ-Proteobacteria, Bacteroidia, Actinobacteria, Acidimicrobiia, and Anaerolineae were the dominant bacteria in all treatments. The dominant genera consisted of Pseudomonas, Sphingobium, Sphingomonas, Cellvibrio, Allorhizobium-Neorhizobium-Pararhizobium-Rhizobium, Flavobacterium, and Algoriphagus (relative abundance>1%). Correlation analysis of environmental factors showed that the composition of the rhizosphere bacterial community was strongly correlated with pH, EC, total nitrogen, and total phosphorus content. The abundances of pathogenic bacteria and antibiotic resistance genes were 103-107 copies·g-1 and 104-108 copies·g-1, respectively. There were significant differences in the detection levels of pathogens and antibiotic resistance genes. Bioorganic fertilizer, loosening soil essence, and corn vinasse significantly increased the abundances of some antibiotic resistance genes, whereas humic acid and corn vinasse significantly decreased the abundances of Pseudomonas syringae, Ralstonia solanacearum, and total coliforms (P<0.05). A significant correlation was found between pathogens (Arcobacter, Bacillus cereus, Pantoea agglomerans, and Fecal bacteroidetes) and antibiotic resistance genes (tetA, tetB, tetO, tetQ, sul1, ermB, and ermC). In conclusion, while monitoring pathogens and antibiotic resistance genes in the agricultural environment under reclaimed water irrigation, attention should be paid to the rational application of soil amendments to avoid exacerbating the spread of biological contamination.

[Effects of Selected Biochars Application on the Microbial Community Structures and Diversities in the Rhizosphere of Water Spinach (Ipomoea aquatica Forssk.) Irrigated with Reclaimed Water].

The utilization of reclaimed water is one of the most important ways of alleviating the shortage of water resources for agricultural irrigation. As an effective disposal method for biomass waste, biochar has been widely used in the improvement and remediation of agricultural environments. However, few studies have been performed on the effects of biochar application on microbial community structures and pathogen abundances in rhizosphere soils irrigated with reclaimed water. Based on a pot experiment, high throughput sequencing technology and quantitative polymerase chain reaction (PCR) methods were used to investigate the effects of different biochars on the microbial community structure and diversity and pathogen abundance of rhizosphere soils irrigated with reclaimed water. The results showed that four different types of biochars had different effects on the soil nutrient status. Rice hull-derived biochar and rice straw-derived biochar resulted in significantly increased soil pH with reclaimed water irrigation, while peanut shell-derived biochar, rice straw-derived biochar, and wheat straw-derived biochar significantly increased EC values (P<0.05). The Sobs index, Shannon index, and Chao1 index of bacterial community in the rhizosphere soil significantly increased with rice straw-derived biochar treatment, while the Simpson index significantly decreased by adding peanut shell-derived biochar, rice hull-derived biochar, and wheat straw-derived biochar (P<0.05). There were differences in the relative abundances of bacterial communities in rhizosphere soils under different treatments. The dominant taxonomic groups at the phylum level were Proteobacteria, Actinobacteria, Chloroflexi, Bacteroidetes, and Acidobacteria. The dominant genera included Pseudomonas, Rheinheimera, Arthrobacter, Sphingomonas, and Aeromonas (relative abundance>5%). Redundancy (RDA) and heatmap analyses showed that the diversities and compositions of bacterial communities in rhizosphere soils in different treatments were closely related to soil EC values, organic matter, total nitrogen, and cadmium contents. Biochar application had no significant effect on the abundances of Aeromonas hydrophila and Bacillus cereus. Rice straw-derived biochar and peanut shell-derived biochar could significantly reduce the γ-Proteobacteria, while rice hull-derived biochar and wheat straw-derived biochar could significantly reduce the relative abundance of AOA (P<0.05). In conclusion, there were no obvious negative effects observed from reclaimed water irrigation on soil quality. Biochar application not only significantly improved the physicochemical properties of the soil, but also impacted the bacterial community structure and the abundance of the functional bacteria in the rhizosphere soil, which was closely related to the soil properties.

[Effect of Different Reclaimed Water Irrigation Methods on Bacterial Community Diversity and Pathogen Abundance in the Soil-Pepper Ecosystem].

Reclaimed water is considered to be a reasonable and sustainable alternative water resource to improve water resource layout and mitigate the shortage of traditional water resources. Its use in irrigation will cause changes in the microbial community structure and opportunistic pathogen abundance in soils and crops, but few studies have been conducted on this subject. Peppers were used as the research subjects, and the treatments were direct irrigation of reclaimed water, mixed irrigation with freshwater and reclaimed water, rotated irrigation with freshwater and reclaimed water, with potable water irrigation as the control. The effects of different irrigation methods of reclaimed water on the soil physicochemical properties were analyzed through a pot experiment. Furthermore, changes in bacterial community and opportunistic pathogen abundance in pepper fruit and the rhizosphere under reclaimed water irrigation conditions were investigated based on high-throughput sequencing technology and quantitative PCR methods. The results showed that direct irrigation with reclaimed water increased soil EC and decreased soil pH. 16S rDNA high-throughput sequencing showed that Proteobacteria, Bacteroides, Actinobacteria, and Firmicutes were present in both pepper fruit and the rhizosphere at phylum level, and the most dominant genera (Pantoea, Pseudomonas, Sphingomonas, Sphingopyxis, Luteimonas, and Mariniflexile) were greatly affected by reclaimed water irrigation methods. Quantitative PCR results indicated that the influence of reclaimed water irrigation on the distribution and abundance of pathogenic bacteria in the soil-pepper system was different, and the abundance of Legionella spp. in pepper fruit and Pseudomonas syringae in the rhizosphere increased with reclaimed water irrigation. Our results indicated that the reclaimed water was suitable for agricultural irrigation, but different reclaimed water irrigation methods may introduce different degrees of microbial contamination. In addition, attention must be given to some opportunistic pathogens and phytopathogens.

[Effects of Different Levels of Irrigation with Reclaimed Water on Soil Enzyme Activity and Distribution of Thermotolerant Coliforms].

This study aimed to discern the effect of different levels of irrigation with reclaimed water on soil enzyme activities and on the distribution of thermotolerant coliforms. The effects of two irrigation water qualities (reclaimed water and tap water) and two irrigation levels (full irrigation and insufficient irrigation) on soil enzyme activity and distribution of thermotolerant coliforms were studied through indoor soil column irrigation. Results indicated that:① Reclaimed water irrigation increased soil urease and invertase activity compared with tap water under the same irrigation conditions. Compared with insufficient irrigation with reclaimed water, full irrigation with reclaimed water significantly increased soil invertase activity and deep soil catalase activity. ② At the same irrigation level, the number of thermotolerant coliforms in soil significantly increased with reclaimed water irrigation. The number of thermotolerant coliforms in soil under full irrigation with reclaimed water was significantly higher than with insufficient irrigation using reclaimed water. The number of thermotolerant coliforms in surface soil of the four irrigated treatments was significantly higher than in the deep soil layer, and with progression of irrigation, the number of thermotolerant coliforms in the 0-60 cm soil layer decreased. ③ There was a significant positive correlation between the quantity of thermotolerant coliforms and the activity of soil urease and invertase, and between the quantity of thermotolerant coliforms and the activity of soil catalase. Reclaimed water therefore promoted enzyme activity that can promote transformation of C and N in soil. Reasonable irrigation with reclaimed water can therefore effectively control the number of thermotolerant coliforms in soil.

[Impact on the Microbial Biomass and Metabolic Function of Carbon Source by Black Soil During Rice Cultivation].

The effects of rice cultivation to the black soil microbial communities, which the experimentation area of Shuangyang District Agricultural Technology Extension Station in Changchun city, Jilin Province of northeastern China, were studied by using the method of phospholipid fatty acids and Biolog ECO-microplate culture. Results showed that the content of organic matter in space was the highest, fewer in the field, and the minimum in the rhizosphere, that change trend of total nitrogen and organic matter was similar in soil. The quantity of organic matter in summer sample was the highest. The microbial fun6tional diversity was significantly higher in summer than that in spring and autumn and showed no significant difference between spring and autumn. For summer and the lowest in winter, Shannon-Wiener index and Pielou index of the space were higher than the field and the rhizosphere. The time of microbial growth into the stable period and peak value of the average well color development were different in all samples, that the time was 216 h, 192 h, 216 h, 120 h, which varied from 0.52-0.84, 0.82-1.28, 0.40-0.84, 0.05-0.48, respectively. The result showed that the time of microbial growth into the stable period was similar in spring and autumn, the highest was in summer and the lowest was in winter. Above all, these results would provide more important characteristics of microbial features in the degradation and restoration process of the quality of the black soil habitat scientifically.

[Running Condition and Bacterial Community Associated with the Partial Nitritation System].

In partial nitritation process, the sludge concentration was higher than 4000 mg x L(-1), DO was less than 0.2 mg x L(-1), the temperature of 15-29 degrees C, the reactor HRT of 4.6 h, and the circulation ratio was 75%, not only the microbial biomass reached the highest value but also the ratio of nitrite to ammonium of the effluent could be controlled at approximate 1.0 by adjusting the dissolved oxygen concentration. A quantitative PCR and the 16S rRNA genes clone library results demonstrated that ammonia-oxidizing bacteria were the dominant among nitrite-oxidizing bacteria at low dissolve oxygen concentration, that the bacteria promoted the running of the partial nitritation. Compared with conventional nitrification systems, in partial nitritation process, Nitrobacter and Nitrospira could not be detected among the system, and abundance of Planctomycetes was higher in partial nitritation process than that of conventional nitrification systems. Phylogenetic analysis of bacterial amoA gene sequences suggested that ammonia-oxidizing bacteria of the partial nitritation system was dominated by Nitrosomonas sequences, so partial nitritation is to be feasible technically and economically.

[Impact on the microbial community of municipal sewage in the anammox system during the cooling process].

Anaerobic ammonium oxidation is an important part of the biological nitrogen removal process, and the performance of the process is determined by the microbial community structure. Low-temperature anaerobic ammonium oxidation technology has good prospects for saving a lot of energy, and anaerobic ammonium oxidation bacteria play a vital role in the removal of total nitrogen from waste water. To explore the microbial community structure changes of anammox reactor in sewage treatment during the cooling process (from 30 degrees C to 20 degrees), the total amount of the microbial, the quantity of anaerobic ammonium oxidation and the change of functional microbial community were investigated in a sewage treatment process using the phospholipid fatty acid method (PLFA), quantitative PCR and the clone library of bacterial 16S rRNA. The PLFAs results showed that the total amount of microbial was first decreased and then gradually increased with the running time, when the temperature dropped from 30 degrees C to 20 degrees C, and the NH4+ -N content in the effluent of the system was decreased. The quantitative PCR results showed that 16S rRNA gene copies of anammox bacteria increased from 1.19 x 10(8) copies x mL(-1) to 1.86 x 10(8) copies x mL(-1) in the wastewater. The PCR-DGGE results showed that when the temperature decreased, the anammox bacteria were further enriched. A shift of anammox bacteria community from single Candidatus Kuenenia sp. to a combination of Candidatus Brocadia sp. and Candidatus Kuenenia sp. was observed.

[Analysis of Microbial Community in the Membrane Bio-Reactor (MBR) Rural Sewage Treatment System].

Uncontrolled release and arbitrary irrigation reuse of rural wastewater may lead to water pollution, and the microbial pathogens could threaten the safety of freshwater resources and public health. To understand the microbial community structure of rural wastewater and provide the theory for microbial risk assessment of wastewater irrigation, microbial community diversities in the Membrane Bio-Reactor (MBR) process for rural wastewater treatment was studied by terminal restriction fragment length polymorphism (T-RFLP) and 16S rDNA gene clone library. Meanwhile, changes of Arcobacter spp. and total bacteria before and after treatment were detected through real-time quantitative PCR. The clone library results showed that there were 73 positive clones included Proteobacteria (91. 80%), Firmicutes (2. 70%), Bacteroidetes (1. 40%), and uncultured bacteria (4. 10%) in the untreated wastewater. The typical pathogenic genus Arcobacter belonging to e-Proteobacteria was the dominant component of the library, accounting for 68. 5% of all clones. The main groups and their abundance in different treatments were significantly distinct. The highest values of species abundance (S), Shannon-Wiener (H) and Evenness (E) were observed in the adjusting tank, which were 43. 0, 3. 56 and 0. 95, respectively. The real-time quantitative PCR results showed that the copy number of Arcobacter spp. was (1. 09 ± 0. 064 0) x 10(11) copies.L-1 in the untreated sewage, which was consistent with the result of 16S rDNA gene clone library. Compared to untreated wastewater, bacterial copy number in the treated effluent decreased 100 to 1 000 times, respectively, suggesting that MBR treatment system could remove the microbial quantity in such scale. In the recycled water, the physicochemical parameters and indicator bacteria met the water quality standard of farmland irrigation. However, further research is needed to estimate the potential health risks caused by residual pathogenic microorganisms in future.

[Influence of Mirabilis jalapa Linn. Growth on the Microbial Community and Petroleum Hydrocarbon Degradation in Petroleum Contaminated Saline-alkali Soil].

In order to explore the effect of Mirabilis jalapa Linn. growth on the structure characteristics of the microbial community and the degradation of petroleum hydrocarbon (TPH) in the petroleum-contaminated saline-alkali soil, Microbial biomass and species in the rhizosphere soils of Mirabilis jalapa Linn. in the contaminated saline soil were studied with the technology of phospholipid fatty acids (PLFAs) analysis. The results showed that comparing to CK soils without Mirabilis jalapa Linn., the ratio of PLFAs species varied were 71. 4%, 69. 2% and 33. 3% in the spring, summer and autumn season, respectively. In addition, there was distinct difference of the biomasses of the microbial community between the CK and rhizosphere soils and among the difference seasons of growth of Mirabilis jalapa Linn.. Compare to CK soil, the degradation rates of total petroleum hydrocarbon (TPH) was increased by 47. 6%, 28. 3%, and 18. 9% in spring, summer, and autumn rhizosphere soils, respectively. Correlation analysis was used to determine the correlation between TPH degradation and the soil microbial community. 77. 8% of the total soil microbial PLFAs species showed positive correlation to the TPH degradation (the correlation coefficient r > 0), among which, 55. 6% of PLFAs species showed high positive correlation(the correlation coefficient was r≥0. 8). In addition, the relative content of SAT and MONO had high correlation with TPH degradation in the CK sample soils, the corelation coefficient were 0. 92 and 0. 60 respectively; However, the percent of positive correlation was 42. 1% in the rhizosphere soils with 21. 1% of them had high positive correlation. The relative content of TBSAT, MONO and CYCLO had moderate or low correlation in rhizosphere soils, and the correlation coefficient were 0. 56, 0. 50, and 0. 07 respectively. Our study showed that the growth of mirabilis Mirabilis jalapa Linn. had a higher influence on the species and biomass of microbial community in the rhizosphere soils, and the results will provide a basis theory for the research of phytoremediation petroleum contaminated saline soil.