Youzhi Feng

Autotrophic growth of nitrifying community in an agricultural soil

The two-step nitrification process is an integral part of the global nitrogen cycle, and it is accomplished by distinctly different nitrifiers. By combining DNA-based stable isotope probing (SIP) and high-throughput pyrosequencing, we present the molecular evidence for autotrophic growth of ammonia-oxidizing bacteria (AOB), ammonia-oxidizing archaea (AOA) and nitrite-oxidizing bacteria (NOB) in agricultural soil upon ammonium fertilization. Time-course incubation of SIP microcosms indicated that the amoA genes of AOB was increasingly labeled by 13CO2 after incubation for 3, 7 and 28 days during active nitrification, whereas labeling of the AOA amoA gene was detected to a much lesser extent only after a 28-day incubation. Phylogenetic analysis of the 13C-labeled amoA and 16S rRNA genes revealed that the Nitrosospira cluster 3-like sequences dominate the active AOB community and that active AOA is affiliated with the moderately thermophilic Nitrososphaera gargensis from a hot spring. The higher relative frequency of Nitrospira-like NOB in the 13C-labeled DNA suggests that it may be more actively involved in nitrite oxidation than Nitrobacter-like NOB. Furthermore, the acetylene inhibition technique showed that 13CO2 assimilation by AOB, AOA and NOB occurs only when ammonia oxidation is not blocked, which provides strong hints for the chemolithoautotrophy of nitrifying community in complex soil environments. These results show that the microbial community of AOB and NOB dominates the nitrification process in the agricultural soil tested.

Long-Term Balanced Fertilization Decreases Arbuscular Mycorrhizal Fungal Diversity in an Arable Soil in North China Revealed by 454 Pyrosequencing

A balanced fertilization can increase crop yields partly due to stimulated microbial activities and growths. In this study, we investigated arbuscular mycorrhizal fungi (AMF) in arable soils to determine the optimal practices for an effective fertilization. We used pyrosequencing-based approach to study AMF diversity, as well as their responses to different long-term (>20 years) fertilizations, including OM (organic manure) and mix chemical fertilizers of NP (nitrogen-phosphorus), NK (nitrogen-potassium), and NPK (nitrogen-phosphorus-potassium). Results revealed that 124,998 of 18S rRNA gene fragments were dominated by Glomeromycota with 59,611 sequences, generating 70 operational taxonomic units (OTUs), of which the three largest families were Glomeraceae, Gigasporaceae and Acaulosporaceae. In Control and NK plots, AMF diversity and richness significantly decreased under long-term P fertilizations, such as NP, NPK, and OM. Concomitantly, the AMF community structure shifted. Supported by canonical correspondence analysis, we hereby propose that long-term balanced fertilization, especially P fertilizer with additional N fertilizer, helps the build-up of soil nutrients. Consequently, some AMF community constituents are sacrificed, propelled by the self-regulation of plant-AMF-microbes system, resulting in an agro-ecosystem with a better sustainability. This knowledge would be valuable toward better understandings of AMF community in agro-ecosystem, and long-term ecosystem benefits of the balanced fertilization.

The Role of Metal Nanoparticles in Influencing Arbuscular Mycorrhizal Fungi Effects on Plant Growth

A knowledge gap still remains concerning the in situ influences of nanoparticles on plant systems, partly due to the absence of soil microorganisms. Arbuscular mycorrhizal fungi (AMF) can form a mutualistic symbiosis with the roots of over 90% of land plants. This investigation sought to reveal the responses of mycorrhizal clover (Trifolium repens) to silver nanoparticles (AgNPs) and iron oxide nanoparticles (FeONPs) along a concentration gradient of each. FeONPs at 3.2 mg/kg significantly reduced mycorrhizal clover biomass by 34% by significantly reducing the glomalin content and root nutrient acquisition of AMF. In contrast, no negative effects of AgNPs at concentrations over 0.1 mg/kg were observed; however, AgNPs at 0.01 mg/kg inhibited mycorrhizal clover growth. In response to the elevated AgNPs content, the ability of AMF to alleviate AgNPs stress (via increased growth and ecological behaviors) was enhanced, which decreased Ag content and the activities of antioxidant enzymes in plants. These results were further supported by X-ray microcomputed tomography. Our findings suggest that in soil ecosystem, the influence of nanometals on plant systems would be more complicated than expected, and more attention should be focused on plant responses in combination with those of soil microorganisms.

Biosorption and Bioreduction of Trivalent Aurum by Photosynthetic Bacteria Rhodobacter capsulatus

Biosorption has been shown to be an eco-friendly approach to remove heavy metal ions. In this study, the photosynthetic bacteria Rhodobacter capsulatus was screened and found to have strong ability to adsorb Au(III). The maximum specific uptake of living cells was over 92.43 mg HAuCl4/g dry weight of cell in the logarithmic phase. Biosorpion ability would be enhanced by an acidic environment. As the main cations, during biosorption the quantity of Mg2+ exchanged was more than Na+. Biosorbed Au(III) could be reduced by carotenoid and enzymes embedded and/or excreted by R. capsulatus, which might be the mechanism of photosynthtic bacteria metal tolerance.

Different responses of soil microbial metabolic activity to silver and iron oxide nanoparticles

The knowledge regarding the effects of metal or metal oxide nanoparticles on soil microbial metabolic activity and key ecological functions is limited, relative to the information about their species diversity. For this reason, the responses of soil microbial metabolic activity to silver (AgNPs) and iron oxide (FeONPs) nanoparticles, along concentration gradients of each, were evaluated by microcalorimetry and soil nitrification potential. The changes in abundances of bacteria, eukaryotes and ammonia-oxidizing bacteria were measured by real time quantitative PCR. It was found that AgNP (at 0.1, 1 and 10 mg kg(-1) soil) amendments decreased soil microbial metabolic activity, nitrification potential and the abundances of bacteria and ammonia-oxidizing bacteria; on the contrary, FeONPs had the positive effects on soil microbial metabolic activity (at 1 and 10 mg kg(-1) soil) and soil nitrification potential (at 0.1 and 1 mg kg(-1) soil). Specific microbial metabolic activity and specific nitrification potential further revealed that metal or metal oxide nanoparticles could change the C and N cycles of the agricultural soil through influencing soil microbial metabolism. These findings could deepen the understanding of the influence of NPs on soil microorganisms and their driven soil ecology process.

The contrasting responses of soil microorganisms in two rice cultivars to elevated ground-level ozone

Although elevated ground-level O₃ has a species-specific impact on plant growth, the differences in soil biota responses to O₃ pollution among rice cultivars are rarely reported. Using O₃ Free-Air Concentration Enrichment, the responses of the rhizospheric bacterial communities in the O₃-tolerant (YD6) and the O₃-sensitive (IIY084) rice cultivars to O₃ pollution and their differences were assessed by pyrosequencing at rice tillering and anthesis stages. Elevated ground-level O₃ negatively influenced the bacterial community in cultivar YD6 at both rice growth stages by decreasing the bacterial phylogenetic diversities and response ratios. In contrast, in cultivar IIY084, the bacterial community responded positively at the rice tillering stage under O₃ pollution. However, several keystone bacterial guilds were consistently negatively affected by O₃ pollution in two rice cultivars. These findings indicate that continuously O₃ pollution would negatively influence rice agroecosystem and the crop cultivar is important in determining the soil biota responses to elevated O₃.

Elevated ground-level O3 negatively influences paddy methanogenic archaeal community

The current knowledge regarding the effect of global climate change on rice-paddy methane (CH4) emissions is incomplete, partly because information is limited concerning the mechanism of the microbial response to elevated ground-level ozone (O3). A field experiment was conducted in the China Ozone Free-Air Concentration Enrichment facility in a rice–wheat rotation system to investigate the responses of methanogenic archaeal communities to elevated ground-level O3 by culture-independent and -reliant approaches. We found that elevated ground-level O3 inhibited methanogenic activity and influenced the composition of paddy methanogenic communities, reducing the abundance and diversity of paddy methanogens by adversely affecting dominant groups, such as aceticlastic Methanosaeta, especially at the rice tillering stage. Our results indicated that continuously elevated ground-level O3 would negatively influence paddy methanogenic archaeal communities and its critical ecological function. These findings will contribute to a comprehensive understanding of the responses and feedbacks of paddy ecosystems to global climate change.

The effect of γ-Fe2O3 nanoparticles on Escherichia coli genome

Extensive production and application of γ-Fe(2)O(3) magnetic nanoparticles (MNPs) has increased their potential risk on environment and human health. This report illustrates a genetic impact of γ-Fe(2)O(3) magnetic nanoparticles (MNPs) on Escherichia coli (E. coli). After 3000-generation incubation with MNPs addition, obvious genomic variations were revealed by using repetitive extragenic palindromic PCR (rep-PCR) DNA fingerprint technique. The physicochemical interactions between MNPs and bacteria could be responsible for such genomic responses. It was revealed that Fe(3+) concentration increased in the medium. Transmission electronic microscopy (TEM) and flow cytometry (FCM) analysis consistently demonstrated the occurrences of adsorption and membranes-internalization of MNPs outside and inside cells. Both increased Fe(3+) ion and the uptake of MNPs facilitated Fe binding with proteins and DNA strands, resulting in enhancing the mutation frequency of E. coli. Our results would be of great help to assessing the potential impact of MNPs on human and environment.

Prevalence of antibiotic resistance genes in soils after continually applied with different manure for 30 years

A 30year field experiment with a wheat-soybean cropping system were performed to compare the long-term anthropogenic influence on soil ARGs. Compared with chemical fertilization, the occurrence of 38 ARGs and the abundance of seven ARGs (tetL, tetB(P), tetO, tetW, sul1, ermB, and ermF) were significantly increased by long term exposure of pig manure. However, application of wheat straw and cow manure not substantially affected the abundance of ARGs except cow manure increased the abundance of tetM and tetW to a detectable level. Relative abundance of Firmicutes, Gammaproteobacteria and Bacteroidetes and ARGs observed in the soil were significantly correlated. Integrase gene I1 (intI1) is commonly linked to genes conferring resistance to antibiotics, it was significantly increased in pig manure treated soils and showed a high positive correlation with the abundance of ARGs which were significantly affected by pig manure. Concentrations of Cu, Zn and Pb were also increased in manure treated soil and positive correlated with the relative abundance of intI1 and most of the ARGs. These results indicated that long-term animal manure application to soils has polluted the soil, especially for pig manure, and it should be scrutinized as part of future stewardship programs.

Elevated Ground-Level O 3 Changes the Diversity of Anoxygenic Purple Phototrophic Bacteria in Paddy Field

The knowledge of the impact of elevated ground-level O3 below ground the agro-ecosystem is limited. A field experiment in China Ozone Free-Air Concentration Enrichment (FACE-O3) facility on a rice–wheat rotation system was carried out to investigate responses of anoxygenic phototrophic purple bacteria (AnPPB) to elevated ground-level O3. AnPPB community structures and sizes in paddy soil were monitored by molecular approaches including PCR–DGGE and real-time quantitative PCR based upon the pufM gene on three typical rice growth stages. Repetitive sequence-based PCR (rep-PCR) in combination with culture-reliant method was conducted to reveal changes in genotypic diversity. Elevated ground-level O3 statistically reduce AnPPB abundance and percentage in total bacterial community in flooded rice soil via decreasing their genotypic diversity and metabolic versatility. Concomitantly, their community composition changed after rice anthesis stage under elevated ground-level O3. Our results from AnPPB potential responses imply that continuously elevated ground-level O3 in the future would eventually harm the health of paddy ecosystem through negative effect on soil microorganisms.