Detection and analysis of sulfur metabolism genes in Sphaerotilus natans subsp. sulfidivorans representatives

The lithotrophic capacity of the betaproteobacteria Sphaerotilus natans subsp. sulfidivorans was confirmed at the genetic level: functional genes of sulfur metabolism were detected (aprBA, soxB, and sqr, coding for adenylyl phosphosulfate reductase, thiosulfate-cleaving enzyme, and sulfide:quinone oxidoreductase, respectively), and the expression of aprA and soxB genes was demonstrated. An evolutionary scenario for soxB genes in Sphaerotilus representatives is suggested based on comparative analysis of codon occurrence frequency, DNA base composition (G+C content), and topology of phylogenetic trees. The ancestor bacterium of the Sphaerotilus-Leptothrix group was capable of lithotrophic growth in the presence of reduced sulfur compounds. However, in the course of further evolution, the sulfur metabolism genes, including the soxB gene, were lost by some Sphaerotilus strains. As a result, the lithotrophic Sphaerotilus-Leptothrix group split into two phylogenetic lineages: lithotrophic and organotrophic ones.     

Aluminum tolerance of wheat does not induce changes in dominant bacterial community composition or abundance in an acidic soil

Aims

Aluminum-tolerant wheat plants often produce more root exudates such as malate and phosphate than aluminum-sensitive ones under aluminum (Al) stress, which provides environmental differences for microorganism growth in their rhizosphere soils. This study investigated whether soil bacterial community composition and abundance can be affected by wheat plants with different Al tolerance.

Methods

Two wheat varieties, Atlas 66 (Al-tolerant) and Scout 66 (Al-sensitive), were grown for 60 days in acidic soils amended with or without CaCO₃. Plant growth, soil pH, exchangeable Al content, bacterial community composition and abundance were investigated.

Results

Atlas 66 showed better growth and lower rhizosphere soil pH than Scout 66 irrespective of CaCO₃ amendment or not, while there was no significant difference in the exchangeable Al content of rhizosphere soil between the two wheat lines. The dominant bacterial community composition and abundance in rhizosphere soils did not differ between Atlas 66 and Scout 66, although the bacterial abundance in rhizosphere soil of both wheat lines was significantly higher than that in bulk soil. Sphingobacteriales, Clostridiales, Burkholderiales and Acidobacteriales were the dominant bacteria phylotypes.

Conclusions

The difference in wheat Al tolerance does not induce the changes in the dominant bacterial community composition or abundance in the rhizosphere soils.     

Marichromatium litoris sp. nov. and Marichromatium chrysaorae sp. nov. isolated from beach sand and from a jelly fish (Chrysaora colorata)

Three strains (JA349^T, JA553^T, JA439) of phototrophic sulphur bacteria were isolated from marine habitats of India. 16S rRNA gene sequence of the three strains clustered phylogenetically with members of the genus Marichromatium of the family Chromatiaceae belonging to the class Gammaproteobacteria. All the strains shared highest sequence similarity with the type strains of Marichromatium spp. (96-99% sequence similarity) and the new strains were characterized based on polyphasic taxonomy. Strains JA349^T and JA553^T can be distinguished from closest relative species of the genus Marichromatium with respect to distinct differences in cellular polar lipids, fatty acids and carbon/nitrogen sources utilization. Both strains were distinctly related (<50% based on DNA-DNA hybridization) with the type strains of the genus Marichromatium. Multilocus Sequence Analysis (MLSA) of the concatenated five protein coding genes (fusA, pufM, dnaK, recA, soxB) along with internal transcribed spacer (ITS; 16S-23S rRNA) had sequence similarity of less than 92% with the type strains of Marichromatium spp. Distinct phenotypic, chemotaxonomic and molecular differences allow the separation of strains JA349^T and JA553^T into new species of the genus Marichromatium for which, we propose the names Marichromatium litoris sp. nov. and Marichromatium chrysaorae sp. nov., respectively.     

Response of archaeal communities in the rhizosphere of maize and soybean to elevated atmospheric CO2 concentrations

Background

Archaea are important to the carbon and nitrogen cycles, but it remains uncertain how rising atmospheric carbon dioxide concentrations ([CO₂]) will influence the structure and function of soil archaeal communities.

Methodology/Principal Findings

We measured abundances of archaeal and bacterial 16S rRNA and amoA genes, phylogenies of archaeal 16S rRNA and amoA genes, concentrations of KCl-extractable soil ammonium and nitrite, and potential ammonia oxidation rates in rhizosphere soil samples from maize and soybean exposed to ambient (∼385 ppm) and elevated (550 ppm) [CO₂] in a replicated and field-based study. There was no influence of elevated [CO₂] on copy numbers of archaeal or bacterial 16S rRNA or amoA genes, archaeal community composition, KCl-extractable soil ammonium or nitrite, or potential ammonia oxidation rates for samples from maize, a model C₄ plant. Phylogenetic evidence indicated decreased relative abundance of crenarchaeal sequences in the rhizosphere of soybean, a model leguminous-C₃ plant, at elevated [CO₂], whereas quantitative PCR data indicated no changes in the absolute abundance of archaea. There were no changes in potential ammonia oxidation rates at elevated [CO₂] for soybean. Ammonia oxidation rates were lower in the rhizosphere of maize than soybean, likely because of lower soil pH and/or abundance of archaea. KCl-extractable ammonium and nitrite concentrations were lower at elevated than ambient [CO₂] for soybean.

Conclusion

Plant-driven shifts in soil biogeochemical processes in response to elevated [CO₂] affected archaeal community composition, but not copy numbers of archaeal genes, in the rhizosphere of soybean. The lack of a treatment effect for maize is consistent with the fact that the photosynthesis and productivity of maize are not stimulated by elevated [CO₂] in the absence of drought.     

Ex-situ enzyme activity and bacterial community diversity through soil depth profiles in penguin and seal colonies on Vestfold Hills, East Antarctica

The soils impacted by sea animal excreta are important sources of nutrients in Antarctic terrestrial ecosystems, and soil microorganisms are the principal drivers of carbon and nitrogen cycling. However, microbial diversity and enzyme activities in these soils have still received little attention. In this paper, we investigated the distribution characteristics of bacterial community in four penguin and seal colony soil profiles collected in East Antarctica, using 16S rDNA-DGGE and real-time quantitative PCR. Soil microbial biomass carbon (C_mic), soil respiration (SR), and enzyme activities involved in carbon, nitrogen, and phosphorus metabolisms were also measured. Overall soil C_mic, SR, enzyme activities, and bacterial abundance decreased with depth. The bacterial abundance had a significant correlation with soil organic carbon and total nitrogen and highly corresponded to the relative content of penguin guano or seal excreta in these soil profiles. The 16S rDNA-DGGE revealed the complicated bacterial community structure in penguin and seal colony soils, and the band richness and dominant bands decreased with soil depth. Cluster analysis of DGGE profiles indicated that bacterial community in those soil profiles were divided into four main categories with the bacterial genetic similarity of 22 %, and the majority of the sequenced bands were Proteobacteria (α, β, γ), Actinobacteria, Bacteroidetes, Deinococcus-Thermus, Chloroflexi, and Firmicutes. Our results indicated that the deposition of penguin guano or seal excreta, which caused the variability in soil soil organic carbon, total nitrogen, pH, and soil moisture, might have an important effect on the vertical distribution pattern of bacterial abundance and diversity in Antarctic soil profiles.     

Rhizosphere soil bacterial communities and nitrogen cycling affected by deciduous and evergreen tree species

Deciduous and evergreen trees differ in their responses to drought and nitrogen (N) demand. Whether or not these functional types affect the role of the bacterial community in the N cycle during drought remains uncertain. Two deciduous tree species (Alnus cremastogyne, an N₂‐fixing species, and Liquidambar formosana) and two evergreen trees (Cunninghamia lanceolata and Pinus massoniana) were used to assess factors in controlling rhizosphere soil bacterial community and N cycling functions. Photosynthetic rates and biomass production of plants, 16S rRNA sequencing and N‐cycling‐related genes of rhizosphere soil were measured. The relative abundance of the phyla Actinobacteria and Firmicutes was higher, and that of Proteobacteria, Acidobacteria, and Gemmatimondaetes was lower in rhizosphere soil of deciduous trees than that of evergreen. Beta‐diversity of bacterial community also significantly differed between the two types of trees. Deciduous trees showed significantly higher net photosynthetic rates and biomass production than evergreen species both at well water condition and short‐term drought. Root biomass was the most important factor in driving soil bacterial community and N‐cycling functions than total biomass and aboveground biomass. Furthermore, 44 bacteria genera with a decreasing response and 46 taxa showed an increased response along the root biomass gradient. Regarding N‐cycle‐related functional genes, copy numbers of ammonia‐oxidizing bacteria (AOB) and autotrophic ammonia‐oxidizing archaea (AOA), N₂ fixation gene (nifH), and denitrification genes (nirK, nirS) were significantly higher in the soil of deciduous trees than in that of the evergreen. Structural equation models explained 50.2%, 47.6%, 48.6%, 49.4%, and 37.3% of the variability in copy numbers of nifH, AOB, AOA, nirK, and nirS, respectively, and revealed that root biomass had significant positive effects on copy numbers of all N‐cycle functional genes. In conclusion, root biomass played key roles in affecting bacterial community structure and soil N cycling. Our findings have important implications for our understanding of plants control over bacterial community and N‐cycling function in artificial forest ecosystems.     

Microbial population index and community structure in saline–alkaline soil using gene targeted metagenomics

Population indices of bacteria and archaea were investigated from saline–alkaline soil and a possible microbe–environment pattern was established using gene targeted metagenomics. Clone libraries were constructed using 16S rRNA and functional gene(s) involved in carbon fixation (cbbL), nitrogen fixation (nifH), ammonia oxidation (amoA) and sulfur metabolism (apsA). Molecular phylogeny revealed the dominance of Actinobacteria, Firmicutes and Proteobacteria along with archaeal members of Halobacteraceae. The library consisted of novel bacterial (20%) and archaeal (38%) genera showing ≤95% similarity to previously retrieved sequences. Phylogenetic analysis indicated ability of inhabitant to survive in stress condition. The 16S rRNA gene libraries contained novel gene sequences and were distantly homologous with cultured bacteria. Functional gene libraries were found unique and most of the clones were distantly related to Proteobacteria, while clones of nifH gene library also showed homology with Cyanobacteria and Firmicutes. Quantitative real-time PCR exhibited that bacterial abundance was two orders of magnitude higher than archaeal. The gene(s) quantification indicated the size of the functional guilds harboring relevant key genes. The study provides insights on microbial ecology and different metabolic interactions occurring in saline–alkaline soil, possessing phylogenetically diverse groups of bacteria and archaea, which may be explored further for gene cataloging and metabolic profiling.     

Salinity controls soil microbial community structure and function in coastal estuarine wetlands

Soil salinity acts as a critical environmental filter on microbial communities, but the consequences for microbial diversity and biogeochemical processes are poorly understood. Here, we characterized soil bacterial communities and microbial functional genes in a coastal estuarine wetland ecosystem across a gradient (~5 km) ranging from oligohaline to hypersaline habitats by applying the PCR-amplified 16S rRNA (rRNA) genes sequencing and microarray-based GeoChip 5.0 respectively. Results showed that saline soils in marine intertidal and supratidal zone exhibited higher bacterial richness and Faith's phylogenetic diversity than that in the freshwater-affected habitats. The relative abundance of taxa assigned to Gammaproteobacteria, Bacteroidetes and Firmicutes was higher with increasing salinity, while those affiliated with Acidobacteria, Chloroflexi and Cyanobacteria were more prevalent in wetland soils with low salinity. The phylogenetic inferences demonstrated the deterministic role of salinity filtering on the bacterial community assembly processes. The abundance of most functional genes involved in carbon degradation and nitrogen cycling correlated negatively with salinity, except for the hzo gene, suggesting a critical role of the anammox process in tidal affected zones. Overall, the salinity filtering effect shapes the soil bacterial community composition, and soil salinity act as a critical inhibitor in the soil biogeochemical processes in estuary ecosystems.     

Microbial Metabolic Potential for Carbon Degradation and Nutrient (Nitrogen and Phosphorus) Acquisition in an Ombrotrophic Peatland

This study integrated metagenomic and nuclear magnetic resonance (NMR) spectroscopic approaches to investigate microbial metabolic potential for organic matter decomposition and nitrogen (N) and phosphorus (P) acquisition in soils of an ombrotrophic peatland in the Marcell Experimental Forest (MEF), Minnesota, USA. This analysis revealed vertical stratification in key enzymatic pathways and taxa containing these pathways. Metagenomic analyses revealed that genes encoding laccases and dioxygenases, involved in aromatic compound degradation, declined in relative abundance with depth, while the relative abundance of genes encoding metabolism of amino sugars and all four saccharide groups increased with depth in parallel with a 50% reduction in carbohydrate content. Most Cu-oxidases were closely related to genes from Proteobacteria and Acidobacteria, and type 4 laccase-like Cu-oxidase genes were >8 times more abundant than type 3 genes, suggesting an important and overlooked role for type 4 Cu-oxidase in phenolic compound degradation. Genes associated with sulfate reduction and methanogenesis were the most abundant anaerobic respiration genes in these systems, with low levels of detection observed for genes of denitrification and Fe(III) reduction. Fermentation genes increased in relative abundance with depth and were largely affiliated with Syntrophobacter. Methylocystaceae-like small-subunit (SSU) rRNA genes, pmoA, and mmoX genes were more abundant among methanotrophs. Genes encoding N₂ fixation, P uptake, and P regulons were significantly enriched in the surface peat and in comparison to other ecosystems, indicating N and P limitation. Persistence of inorganic orthophosphate throughout the peat profile in this P-limiting environment indicates that P may be bound to recalcitrant organic compounds, thus limiting P bioavailability in the subsurface. Comparative metagenomic analysis revealed a high metabolic potential for P transport and starvation, N₂ fixation, and oligosaccharide degradation at MEF relative to other wetland and soil environments, consistent with the nutrient-poor and carbohydrate-rich conditions found in this Sphagnum-dominated boreal peatland.     

Functional Gene Analysis of Freshwater Iron-Rich Flocs at Circumneutral pH and Isolation of a Stalk-Forming Microaerophilic Iron-Oxidizing Bacterium

Iron-rich flocs often occur where anoxic water containing ferrous iron encounters oxygenated environments. Culture-independent molecular analyses have revealed the presence of 16S rRNA gene sequences related to diverse bacteria, including autotrophic iron oxidizers and methanotrophs in iron-rich flocs; however, the metabolic functions of the microbial communities remain poorly characterized, particularly regarding carbon cycling. In the present study, we cultivated iron-oxidizing bacteria (FeOB) and performed clone library analyses of functional genes related to carbon fixation and methane oxidization (cbbM and pmoA, respectively), in addition to bacterial and archaeal 16S rRNA genes, in freshwater iron-rich flocs at groundwater discharge points. The analyses of 16S rRNA, cbbM, and pmoA genes strongly suggested the coexistence of autotrophic iron oxidizers and methanotrophs in the flocs. Furthermore, a novel stalk-forming microaerophilic FeOB, strain OYT1, was isolated and characterized phylogenetically and physiologically. The 16S rRNA and cbbM gene sequences of OYT1 are related to those of other microaerophilic FeOB in the family Gallionellaceae, of the Betaproteobacteria, isolated from freshwater environments at circumneutral pH. The physiological characteristics of OYT1 will help elucidate the ecophysiology of microaerophilic FeOB. Overall, this study demonstrates functional roles of microorganisms in iron flocs, suggesting several possible linkages between Fe and C cycling.     

Effects of elevated CO2 and Pb on the microbial community in the rhizosphere of Pinus densiflora

Rising levels of atmospheric CO₂ may stimulate forest productivity in the future, resulting in increased carbon storage in terrestrial ecosystems. However, heavy metal contamination may interfere with this, though the response is not yet known. In this study, we investigated the effect of elevated CO₂ and Pb contamination on microorganisms and decomposition in pine tree forest soil. Three-year old pine trees (Pinus densiflora) were planted in Pb contaminated soils (500 mg/kg-soil) and uncontaminated soils and cultivated for three months in a growth chamber where the CO₂ concentration was controlled at 380 or 760 mg/kg. Structures of the microbial community were comparatively analyzed in bulk and in rhizosphere soil samples using community-level physiological profiling (CLPP) and 16S rRNA gene PCR-DGGE (denaturing gradient gel electrophoresis). Additionally, microbial activity in rhizospheric soil, growth and the C/N ratio of the pine trees were measured. Elevated CO₂ significantly increased microbial activities and diversity in Pb contaminated soils due to the increase in carbon sources, and this increase was more distinctive in rhizospheric soil than in bulk soils. In addition, increased plant growth and C/N ratios of pine needles at elevated CO₂ resulted in an increase in cation exchange capacity (CEC) and dissolved organic carbon (DOC) of the rhizosphere in Pb contaminated soil. Taken together, these findings indicate that elevated CO₂ levels and heavy metals can affect the soil carbon cycle by changing the microbial community and plant metabolism.     

Roots affect the response of heterotrophic soil respiration to temperature in tussock grass microcosms

Aims and Background

While the temperature response of soil respiration (R_S) has been well studied, the partitioning of heterotrophic respiration (R_H) by soil microbes from autotrophic respiration (R_A) by roots, known to have distinct temperature sensitivities, has been problematic. Further complexity stems from the presence of roots affecting R_H, the rhizosphere priming effect. In this study the short-term temperature responses of R_A and R_H in relation to rhizosphere priming are investigated.

Methods

Temperature responses of R_A, R_H and rhizosphere priming were assessed in microcosms of Poa cita using a natural abundance δ¹³C discrimination approach.

Results

The temperature response of R_S was found to be regulated primarily by R_A, which accounted for 70 % of total soil respiration. Heterotrophic respiration was less sensitive to temperature in the presence of plant roots, resulting in negative priming effects with increasing temperature.

Conclusions

The results emphasize the importance of roots in regulating the temperature response of R_S, and a framework is presented for further investigation into temperature effects on heterotrophic respiration and rhizosphere priming, which could be applied to other soil and vegetation types to improve models of soil carbon turnover.     

Process-based isotope partitioning of winter soil respiration in a subalpine ecosystem reveals importance of rhizospheric respiration

Deep snow in sub-alpine ecosystems may reduce or eliminate soil freezing, thus contributing to the potential for winter soil respiration to account for a significant fraction of annual CO₂ efflux to the atmosphere. Quantification of carbon loss from soils requires separation of respiration produced by roots and rhizosphere organisms from that produced by heterotrophic, decomposer organisms because the former does not result in a net loss of stored carbon. Our objective was to quantify winter soil respiration rates in a sub-alpine forest and meadow, and to partition that flux into its rhizosphere and heterotrophic components. We were particularly interested in comparing early winter soil respiration to late winter/early spring soil respiration of each component because previous work has shown a consistent increase in soil respiration of subalpine systems from early winter to late winter/spring. Field data on the total soil CO₂ flux and its carbon isotope composition were coupled with data from laboratory incubations using a novel process-based stable isotope mixing model implemented in a hierarchical Bayesian framework. We found that soil respiration generally increased from early to later winter and was greatest mid-summer. After correcting for the effect of wind on snowpack δ¹³C–CO₂, the δ¹³C of soil-respired CO₂ varied little over winter, and the contributions of rhizospheric (~35 %) and heterotrophic (~65 %) respiration were relatively constant. The significance of winter respiration from the rhizosphere and apparent coupling of increases in rhizospheric and heterotrophic respiration in late winter are likely to be important for predicting changes in soil carbon in sub-alpine ecosystems.     

Application of targeted metagenomics to explore abundance and diversity of CO2-fixing bacterial community using cbbL gene from the rhizosphere of Arachis hypogaea

Sequestration of CO₂ by autotrophic bacteria is a key process of biogeochemical carbon cycling in soil ecosystem. Rhizosphere is a rich niche of microbial activity and diversity, influenced by change in atmospheric CO₂. Structural changes in rhizosphere composition influence microbial communities and the nutrient cycling. In the present study, the bacterial diversity and population dynamics were established using cbbL and 16S rRNA gene targeted metagenomics approach from the rhizosphere of Arachis hypogaea. A total of 108 cbbL clones were obtained from the rhizospheric soil which revealed predominance of cbbL sequences affiliated to Rhizobium leguminosarum, Bradyrhizobium sp., Sinorhizobium meliloti, Ochrobactrum anthropi and a variety of uncultured cbbL harboring bacteria. The 16S rRNA gene clone library exhibited the dominance of Firmicutes (34.4%), Proteobacteria (18.3%), Actinobacteria (17.2%) and Bacteroidetes (16.1%). About 43% nucleotide sequences of 16S rRNA gene clone library were novel genera which showed < 95% homology with published sequences. Gene copy number of cbbL and 16S rRNA genes, determined by quantitative real‐time PCR (qRT PCR), was 9.38 ± 0.75 × 10⁷ and 5.43 ± 0.79 × 10⁸ (per g dry soil), respectively. The results exhibited bacterial community structure with high bacterial diversity and abundance of CO₂‐fixing bacteria, which can be explored further for their role in carbon cycling, sustainable agriculture and environment management.     

Bacterial diversity in the rhizosphere of maize and the surrounding carbonate‐rich bulk soil

Maize represents one of the main cultivar for food and energy and crop yields are influenced by soil physicochemical and climatic conditions. To study how maize plants influence soil microbes we have examined microbial communities that colonize maize plants grown in carbonate‐rich soil (pH 8.5) using culture‐independent, PCR‐based methods. We observed a low proportion of unclassified bacteria in this soil whether it was planted or unplanted. Our results indicate that a higher complexity of the bacterial community is present in bulk soil with microbes from nine phyla, while in the rhizosphere microbes from only six phyla were found. The predominant microbes in bulk soil were bacteria of the phyla Acidobacteria, Bacteroidetes and Proteobacteria, while Gammaproteobacteria of the genera Pseudomonas and Lysobacter were the predominant in the rhizosphere. As Gammaproteobacteria respond chemotactically to exudates and are efficient in the utilization of plants exudate products, microbial communities associated to the rhizosphere seem to be plant‐driven. It should be noted that Gammaproteobacteria made available inorganic nutrients to the plants favouring plant growth and then the benefit of the interaction is common.     

荒漠草原不同植物根际与非根际土壤养分及微生物量分布特征

通过对宁夏荒漠草原6种地带性优势物种长芒草、蒙古冰草、甘草、牛心朴子、黑沙蒿和苦豆子植物根际与非根际土壤养分和微生物量分布特征进行研究,探讨不同植物根际养分的富集的相关性和差异性。研究结果表明:6种植物根际土壤养分和微生物量均表现出明显的富集效应,根际富集率大小依次为菊科(黑沙蒿)豆科(苦豆子、甘草)禾本科(长芒草、蒙古冰草)萝藦科(牛心朴子);全磷(TP)在根际和非根际中无显著差异(P0.05),其它土壤养分及理化指标在根际中均表现出显著富集(P0.05),土壤养分中以有机碳(SOC)的富集作用最为明显;土壤有效态养分较全量养分对植物根际微小的变化响应更为灵敏;不同荒漠植物根际与非根际SOC与全氮(TN)呈极显著线性关系(P0.01),TN与碱解氮之间呈极显著线性关系(P0.01),TP与有效磷(AP)没有显著的相关性(P0.05)。荒漠植物土壤有效养分在根际存在一定的富集,灌木和豆科植物的根际效应的大于禾本科植物,它们通过降低根际pH值可以提高根际养分,有利于在脆弱环境下对土壤养分的有效利用。     

植物种类对野芷湖湖岸带土壤反硝化作用的影响

反硝化作用是湖岸带有效去除氮素的关键氮循环过程,受到植物、土壤理化性质及微生物等因素的影响。为探究湖岸带不同植物对土壤反硝化作用的影响,通过调查测定野芷湖湖岸带9种常见植物的根际与非根际土壤的反硝化功能基因丰度和反硝化潜势,分析其与土壤理化性质的关系,阐明植物种类影响反硝化作用的机制。研究结果表明:(1)种植植物会改变湖岸带土壤的理化指标,植物根际土的硝态氮(NN)、铵态氮(AN)、总碳(TC)、总氮(TN)和可溶性有机碳(DOC)含量等显著高于非根际土,其中柳树、黄素馨、喜旱莲子草根际土的TC、DOC、TN、NN显著高于其他物种,桂树的根际和非根际土的pH均显著低于其他物种;(2)植物根际土的微生物反硝化功能基因(narG、napA、nirS、nirK和nosZ)丰度显著高于非根际土,其中柳树、黄素馨、喜旱莲子草根际土的基因丰度显著高于其他物种, pH、TN、NN和DOC等对反硝化功能基因丰度影响较大;(3)根际土的反硝化潜势显著高于非根际土,其中柳树、桂树、黄素馨根际土的反硝化潜势显著高于其他物种,黄素馨非根际土的反硝化潜势也较高,pH、DOC、TN以及napA、nirS、norB基因丰度等对土壤的反硝化潜势具有显著影响。本研究说明植物根际微环境更有利于反硝化微生物的生长与繁殖,湖岸带种植植物时可考虑不同的常绿和落叶乔木、灌木、草本植物的合理配置来提高湖岸带的氮素截留能力,以减少由过量氮素进入水生生态系统导致的面源污染。     

Increased Abundance and Transferability of Resistance Genes after Field Application of Manure from Sulfadiazine-Treated Pigs

Spreading manure containing antibiotics in agriculture is assumed to stimulate the dissemination of antibiotic resistance in soil bacterial populations. Plant roots influencing the soil environment and its microflora by exudation of growth substrates might considerably increase this effect. In this study, the effects of manure from pigs treated with sulfadiazine (SDZ), here called SDZ manure, on the abundance and transferability of sulfonamide resistance genes sul1 and sul2 in the rhizosphere of maize and grass were compared to the effects in bulk soil in a field experiment. In plots that repeatedly received SDZ manure, a significantly higher abundance of both sul genes was detected compared to that in plots where manure from untreated pigs was applied. Significantly lower abundances of sul genes relative to bacterial ribosomal genes were encountered in the rhizosphere than in bulk soil. However, in contrast to results for bulk soil, the sul gene abundance in the SDZ manure-treated rhizosphere constantly deviated from control treatments over a period of 6 weeks after manuring, suggesting ongoing antibiotic selection over this period. Transferability of sulfonamide resistance was analyzed by capturing resistance plasmids from soil communities into Escherichia coli. Increased rates of plasmid capture were observed in samples from SDZ manure-treated bulk soil and the rhizosphere of maize and grass. More than 97% of the captured plasmids belonged to the LowGC type (having low G+C content), giving further evidence for their important contribution to the environmental spread of antibiotic resistance. In conclusion, differences between bulk soil and rhizosphere need to be considered when assessing the risks associated with the spreading of antibiotic resistance.     

保护性耕作对东北黑土微生物碳循环功能基因的影响

土壤微生物既参与土壤有机碳的分解也是土壤有机碳转化和固定的驱动者,是影响土壤碳循环和有机碳稳定性的关键因素。然而,保护性耕作（秸秆还田）如何通过调节土壤微生物碳循环功能基因组成来影响土壤CO₂释放的机理尚不明确。因此,依托中国科学院东北地理与农业生态研究所长春保护性耕作观测站,借助鸟枪法宏基因组测序技术,分析了玉米连作系统不同耕作方式（免耕（NT）、秋翻（MP）以及常规耕作（CT））对土壤CO₂释放速率、碳水化合物活性酶（CAZy）、碳循环功能基因（碳固定、甲烷代谢以及碳水化合物代谢）组成的影响。研究表明：基于生长季节土壤CO₂释放速率6年平均值分析发现,生长季前期免耕土壤的平均CO₂释放速率显著低于秋翻和常规耕作,分别比秋翻低28%（5月份）、11%（6月份）和23%（7月份）;比常规耕作低31%（5月份）、19%（6月份）和7%（7月份）。基于CAZy数据库注释结果,发现耕作处理显著影响一些糖苷水解酶（如GH102、GH5_38和GH13_17）、糖基转移酶（如GT39）和多糖裂解酶（如PL17和PL5_1）的基因丰度,与常规耕作相比,秸秆还田的免耕和秋翻处理的这些差异基因的相对丰度较高。基于京都基因与基因组百科全书（KEGG）数据库注释结果,发现耕作方式显著影响土壤碳循环功能基因组成（Adonis,多元方差分析,R²=0.45;P=0.006）,且免耕处理土壤的碳固定、甲烷代谢以及碳水化合物代谢功能基因组成不同于常规耕作和秋翻处理,单独聚为一类。免耕土壤上调的碳固定功能基因的相对丰度（所有上调功能基因相对丰度的平均值）分别比常规耕作和秋翻高17%和11%,而下调的2个功能基因（K01007和K00170）的丰度分别低19%（CT）、21%（MP）和14%（CT）、17%（MP）。免耕土壤上调的甲烷代谢基因相对丰度分别较常规耕作和秋翻高15%和10%;下调基因的丰度分别低13%（CT）和11%（MP）。免耕土壤上调的碳水化合物代谢功能基因丰度较常规耕作和秋翻高23%和14%;下调的基因丰度分别低25%（CT）和18%（MP）。冗余分析（db-RDA）表明土壤容重及土壤水溶性有机碳（DOC）是驱动土壤碳循环功能基因组成差异的主要因子（P<0.05）,且免耕土壤上调的碳固定功能基因（K00625、K01676、K09709、K00925和K14470等）、甲烷代谢基因（K03520、K00830、K10713、K15633和K00625等）和碳水化合物代谢功能基因（K00886、K00830、K01676、K00117和K00114等）与土壤DOC、容重或含水量呈显著正相关。此外,研究发现土壤CO₂释放速率与土壤碳循环功能基因组成显著相关（R²=0.80;P<0.01）,尤其是与一些碳水化合物代谢功能基因显著相关。这些结果说明免耕处理通过影响土壤理化性质改变土壤碳循环过程,且推断免耕秸秆还田和减少干扰的叠加效应通过调节碳循环功能基因组成来提高土壤固碳潜力。