Effects of inoculation with ectomycorrhizal fungi on microbial biomass and bacterial functional diversity in the rhizosphere of Pinus tabulaeformis seedlings

This study investigated the effects of inoculation with three individual ectomycorrhizal (ECM) fungal species on soil microbial biomass carbon and indigenous bacterial community functional diversity in the rhizosphere of Chinese pine (Pinus tabulaeformis Carr.) seedlings under field experimental conditions. The results showed that ECM fungal inoculation significantly increased the ectomycorrhizal colonization compared with non-inoculated seedlings. ECM fungal inoculations have higher soil microbial biomass carbon than that of control, ranging from 49.6 μg C g^?1 dry soil in control to 134.02 μg C g^?1 dry soil in treatment inoculated with Boletus luridus Schaeff ex Fr. Multivariate analyses (PCA) of BIOLOG data revealed that the application of ECM fungi significantly influenced bacterial functional diversity in the rhizosphere of P. tabulaeformis seedlings. The highest average well-color development (AWCD) and functional diversity indices were also observed in treatment inoculated with B. luridus. A wider range of sole carbon sources were utilized by the bacterial community in the rhizosphere of inoculated seedlings. The data gathered from this study provides important information for utilization of ECM fungi in forest restoration project in the Northwestern China. The present study will also significantly broaden our understanding of practical importance in the application of ECM fungal inoculum to promote soil microbial community diversity of soil.     

Rhizosphere bacterial community composition in natural stands of Carex arenaria (sand sedge) is determined by bulk soil community composition

The relative importance of specific plant properties versus soil characteristics in shaping the bacterial community structure of the rhizosphere is a topic of considerable debate. Here, we report the results of a study on the bacterial composition of the rhizosphere of the wild plant Carex arenaria (sand sedge) growing at 10 natural sites in The Netherlands. The soil properties of the sandy soils at these sites were highly disparate, most notably in pH, chloride and organic matter content. Rhizosphere and bulk soil bacterial communities were examined by culture-independent means, namely, 16S rDNA-directed PCR-DGGE profiling. Large differences were observed between the bacterial communities of the different sites for both bulk and rhizosphere soil. Cluster analysis of bacterial profiles revealed that the rhizosphere community of each site was generally more closely related to the bulk soil community of that site rather than to rhizosphere communities of other sites. Hence, bacterial community structure within the rhizosphere of C. arenaria appeared to be determined to a large extent by the bulk soil community composition. This conclusion was supported by a reciprocal planting experiment, where C. arenaria shoots of different sites yielded highly similar rhizosphere communities when planted in the same soil.     

Effects of silver nanoparticles on soil microorganisms and maize biomass are linked in the rhizosphere

Silver nanoparticles hold great promise as effective anti-microbial compounds in a myriad of applications but may also pose a threat to non-target bacteria and fungi in the environment. Because microorganisms are involved in extensive interactions with many other organisms, these partner species are also prone to indirect negative effects from silver nanoparticles.Here, we focus on the effects of nanosilver exposure in the rhizosphere. Specifically, we evaluate the effect of 100 mg kg⁻¹ silver nanoparticles on maize plants, as well as on the bacteria and fungi in the plant's rhizosphere and the surrounding bulk soil. Maize biomass measurements, microbial community fingerprints, an indicator of microbial enzymatic activity, and carbon use diversity profiles are used. Hereby, it is shown that 100 mg kg⁻¹ silver nanoparticles in soil increases maize biomass, and that this effect coincides with significant alterations of the bacterial communities in the rhizosphere. The bacterial community in nanosilver exposed rhizosphere shows less enzymatic activity and significantly altered carbon use and community composition profiles. Fungal communities are less affected by silver nanoparticles, as their composition is only slightly modified by nanosilver exposure. In addition, the microbial changes noted in the rhizosphere were significantly different from those noted in the bulk soil, indicated by different nanosilver-induced alterations of carbon use and community composition profiles in bulk and rhizosphere soil.Overall, microorganisms in the rhizosphere seem to play an important role when evaluating the fate and effects of silver nanoparticle exposure in soil, and not only is the nanosilver response different for bacteria and fungi, but also for bulk and rhizosphere soil. Consequently, assessment of microbial populations should be considered an essential parameter when investigating the impacts of nanoparticle exposure.     

Genetic profiling of bacterial communities from the rhizospheres of ozone damaged Malva sylvestris (Malvaceae)

The phytotoxicity of tropospheric ozone at elevated concentrations can cause severe effects on plant vegetation. This study analyzed whether ozone-damaged plants transmit signals into the soil and thereby affect the structural diversity of bacteria inhabiting the rhizosphere. To generate highly stressful conditions, Malva sylvestris was raised from seeds under ozone exposure. Additionally, the plant age at the onset of ozone-stress was varied. Rhizosphere bacterial communities were analyzed by single-strand conformation polymorphism (SSCP) of PCR amplified partial 16S ribosomal RNA genes. None of these vigorous stress conditions caused a change in the genetic profiles targeting Bacteria with primers hybridizing to highly conserved regions in the 16S rRNA genes, despite visibly ozone-triggered injuries of the leaves and a reduced root biomass. Differences in the SSCP profiles however were observed between plants of different ages. In a more realistic scenario, plants raised in the greenhouse under normal atmospheric conditions were exposed to ozone episodes as they can occur during summer in industrialized countries of the northern hemisphere. To increase the sensitivity of detection, genetic profiles representing Alphaproteobacteria, Actinobacteria and Pseudomonas were analyzed separately in addition to those of Bacteria. None of the profiles showed any ozone-related alterations. In summary, the results of this study indicated that, even when the plants were visibly injured by ozone, the stress was not transmitted to the soil in a way that affected the structural diversity of the dominant rhizosphere bacterial community.     

Release of the recombinant Cry3Bb1 protein of Bt maize MON88017 into field soil and detection of effects on the diversity of rhizosphere bacteria

A three-year experimental field study with a genetically engineered Bt maize (event MON88017) and three conventionally bred cultivars was conducted to quantify the recombinant Cry3Bb1 protein released into soil and detect effects on the diversity of soil bacteria. Protein extraction and an enzyme-linked immunosorbent assay (ELISA) allowed a threshold detection of 0.01 ng Cry3Bb1 g^?1 soil. The maximum amount found in field plots with Bt maize was 1.0 ng Cry3Bb1 g^?1 rhizosphere soil. Average concentrations during the growing seasons varied between years from 0.07 to 0.29 ng g^?1. No accumulation of Cry3Bb1 in soil occurred over the three growing seasons. Four weeks after harvest, the major Cry3Bb1 reservoirs on the field were the remaining root stubbles, but their Cry3Bb1 concentration declined by 98.30–99.99% in the following seven months. During the three consecutive years of study there were never significant differences between the rhizosphere bacterial community structure of the Bt maize and the other cultivars, as detected by cultivation independent profiling of PCR-amplified 16S rRNA genes. The low concentrations of soil extractable Cry3Bb1, its degradation in decaying roots, and the lack of effects on rhizosphere bacteria give no indications of adverse effects of MON88017 cultivation on soil ecology.     

Leaf litter is the main driver for changes in bacterial community structures in the rhizosphere of ash and beech

The rhizosphere and the surrounding soil harbor an enormous microbial diversity and a specific community structure, generated by the interaction between plant roots and soil bacteria. The aim of this study was to address the influences of tree species, tree species diversity and leaf litter on soil bacterial diversity and community composition. Therefore, mesocosm experiments using beech, ash, lime, maple and hornbeam were established in 2006, and sampled in October 2008 and June 2009. Mesocosms were planted with one, three or five different tree species and treated with or without litter overlay.Cluster analysis of DGGE-derived patterns revealed a clustering of 2008 sampled litter treatments in two separated clusters. The corresponding treatments sampled in 2009 showed separation in one cluster. PCA analysis based on the relative abundance of active proteobacterial classes and other phyla in beech and ash single-tree species mesocosm indicated an effect of sampling time and leaf litter on active bacterial community composition. The abundance of next-generation sequencing-derived sequences assigned to the Betaproteobacteria was higher in the litter treatments, indicating a higher activity, under these conditions. The Deltaproteobacteria, Nitrospira and Gemmatimonadetes showed an opposite trend and were more active in the mesocosms without litter. The abundance of alphaproteobacterial sequences was higher in mesocosms sampled in 2009 (P = 0.014), whereas the Acidobacteria were more active in 2008 (P = 0.014). At the family level, we found significant differences of the litter vs. non-litter treated group. Additionally, an impact of beech and ash as tree species on soil bacterial diversity was confirmed by the Shannon and Simpson indices. Our results suggest that leaf litter decomposition in pH-stable soils affect the soil bacterial composition, while tree species influence the soil bacterial diversity.     

Structural differences in the rhizosphere communities of legumes are not equally reflected in community-level physiological profiles

The relationship of structural diversity and differences in the functional potentials of rhizosphere communities of alfalfa, common bean and clover was investigated in microcosms. PCR-SSCP (single strand conformation polymorphism) analysis of 16S rRNA genes revealed significant differences in the composition of the leguminous rhizosphere communities at the shoot stage of plants grown in the same soil. Sequencing of dominant SSCP-bands indicated the presence of plant specific organisms. The partial rRNA gene sequences were related to members of the α- and γ-Proteobacteria, Bacteroidetes and Actinobacteria. Besides the plant species, the soil also affected the structural diversity in rhizospheres. The dominant bacterial populations of alfalfa grown in soils with different agricultural histories were assigned to different taxonomic groups. Addressing the functional potentials, community-level physiological profiles (CLPP) were generated using BIOLOG GN^®. The three leguminous rhizosphere communities could be differentiated by principle component analysis, though the overall analysis indicated that the metabolic potential of all rhizosphere samples was similar. The functional variation examined in rhizospheres of alfalfa was minor in response to the soil origin and was found not to be significant different at different growth stages. The results indicate that similar functional potentials may be provided by structurally different bacterial communities.     

Potato cultivar type affects the structure of ammonia oxidizer communities in field soil under potato beyond the rhizosphere

The effects of plants on the microbiota involved in the oxidation of ammonia in soils have been controversial. Here, we investigated the dynamics in the abundances and community structures of the bacterial and archaeal ammonia oxidizers (AOB and AOA, respectively) in two fields that were cropped with potato. Six different potato cultivars were used, including a genetically-modified one, in a fourfold replicated experimental set-up. On the basis of bulk and rhizosphere soil extracted microbial community DNA, AOB and AOA quantitative PCR as well as PCR-DGGE were performed. In addition, samples were used for the production and analysis of amoA gene fragment based clone libraries. Regardless of sample type (bulk versus rhizosphere soil) and across soils, the population sizes of AOA (of the order 10⁴–10⁸ amoA gene copies g⁻¹ dry soil), were generally higher than those of AOB in the same samples (about 10⁴–10⁵ g⁻¹ dry soil), resulting in ratio's of log-transformed values > 1.0. Whereas the AOB numbers were generally raised in the rhizosphere versus bulk soils in both soils, the opposite was true for the AOA numbers. Moreover, significant effects of cultivar type on both the AOB and AOA community structures were found in both soils, and these extended to beyond the rhizospheres. The effects were found across the whole growth season. Soil type did not significantly affect the community structures of AOA, but had a small effect on the community structure of AOB. Analysis of the structures of the AOB communities revealed a prevalence of AOB subgroups 2, 3a, 3b and 4 in one field soil and of 2 and 4 in the other one. With respect to the AOA, soil/sediment clusters (SS) I, II, III and IV were found to prevail.     

Liming induces growth of a diverse flora of ammonia-oxidising bacteria in acid spruce forest soil as determined by SSCP and DGGE

The autotrophic ammonia-oxidising bacterial (AOB) community composition was studied in acid coniferous forest soil profiles at a site in southwestern Sweden 6 years after liming. Liming caused a significant increase in pH in the organic horizons, while the mineral soil was unaffected. The AOB communities were studied by single-strand conformation polymorphism (SSCP) in parallel with denaturing gradient gel electrophoresis (DGGE) analysis of partial 16S rRNA genes amplified by PCR using primers reported to be specific for β-Proteobacteria AOB, followed by nucleotide sequencing. High genetic diversity of Nitrosospira-like sequences was found in the limed soil profiles, whereas no AOB-like sequences were detected in the control soil at any depth, according to both the SSCP and DGGE analyses. This clearly showed that liming induced growth of a diverse flora of AOB at this forest site. Both Nitrosospira cluster 2 and cluster 4 sequences were present in the limed soil profiles, regardless of soil pH, but we found a higher number of sequences affiliated with cluster 4. The high lime dose seemed to affect the AOB community more than the low dose, and its effects reached deeper into the soil profile. Seven different Nitrosospira-like sequences were found 10 cm under the litter layer in the soil limed with the high dose, but only two in the soil amended with the low lime dose.     

Soil microbial community response to controlled-release urea fertilizer under zero tillage and conventional tillage

Soil microorganisms mediate many important biological processes for sustainable agriculture. The effect of a polymer-coated controlled-release urea (CRU, ESN^®) on soil microbial communities was studied at six sites across western Canada from 2004 to 2006. Fertilizer treatments were CRU, urea and an unfertilized control. Timing of fertilizer application (fall vs. spring) was studied in 9 of the 18 site-years (combinations of sites and years). Wheat (Triticum aestivum L.), canola (Brassica napus L.) and barley (Hordeum vulgare L.) were grown in rotation at five sites, and silage corn (Zea mays L.) was grown in all 3 years at one site, under conventional tillage (CT) or zero tillage (ZT). The fertilizers were side-banded at 50–60 kg N ha⁻¹ for wheat, barley and canola, and broadcast at 150 kg N ha⁻¹ for corn. Microbial biomass C (MBC) and bacterial functional diversity and community-level physiological profiles (CLPPs) were determined at about the flowering stage of each crop. In situ CO₂ evolution (soil respiration) was measured, and microbial metabolic quotient (qCO₂) determined, at one site in 2 years. In the rhizosphere, fertilizer effects on MBC and functional diversity were observed in 1 and 5 of 18 site-years, respectively; and in bulk soil in 4 site-years each. These effects were usually positive relative to the control. CRU increased MBC or functional diversity more than urea in 3 site-years, but the opposite was observed in 1 site-year. Time of fertilizer application affected MBC in 1, and functional diversity in 2, of 9 site-years in the rhizosphere, and no effects were observed in bulk soil. Fall-applied fertilizer increased MBC more than spring-applied fertilizer, but the opposite was observed for functional diversity. Tillage affected MBC and functional diversity in 4 and 5 of 18 site-years, respectively, in the rhizosphere, and in 3 and 4 site-years in bulk soil. Tillage effects were usually in favour of ZT. There were no treatment effects on CO₂ evolution, but an interactive effect of fertilizer and tillage on qCO₂ was observed in 1 year when qCO₂ in the control treatment was greater than that in either fertilizer treatment under CT, but urea increased qCO₂ relative to the control under ZT. Shifts in CLPPs were sometimes observed where the treatment effects described above were not significant. Notwithstanding the limitations of culture-dependent CLPPs, most fertilizer effects on soil microbiological properties were not statistically significant. Therefore, these fertilizers probably did not adversely affect most soil biological processes.     

Effects of tree identity dominate over tree diversity on the soil microbial community structure

This study investigated the possible effects of tree species diversity and identity on the soil microbial community in a species-rich temperate broad-leaved forest. For the first time, we separated the effects of tree identity and tree species diversity on the link between above and belowground communities in a near-natural forest. We established 100 tree clusters consisting of each three tree individuals represented by beech (Fagus sylvatica L.), ash (Fraxinus excelsior L.), hornbeam (Carpinus betulus L.), maple (Acer pseudoplatanus L.), or lime (Tilia spec.) at two different sites in the Hainich National Park (Thuringia, Germany). The tree clusters included one, two or three species forming a diversity gradient. We investigated the microbial community structure, using phospholipid fatty acid (PLFA) profiles, in mineral soil samples (0–10 cm) collected in the centre of each cluster.The lowest total PLFA amounts were found in the pure beech clusters (79.0 ± 23.5 nmol g⁻¹ soil dw), the highest PLFA amounts existed in the pure ash clusters (287.3 ± 211.3 nmol g⁻¹ soil dw). Using principle components analyses (PCA) and redundancy analyses (RDA), we found only for the variables ‘relative proportion of beech trees’ and ‘living lime fine root tips associated with ectomycorrhiza’ a significant effect on the PLFA composition. The microbial community structure was mainly determined by abiotic environmental parameters such as soil pH or clay content. The different species richness levels in the clusters did not significantly differ in their total PLFA amounts and their PLFA composition. We observed a tendency that the PLFA profiles of the microbial communities in more tree species-rich clusters were less influenced by individual PLFAs (more homogenous) than those from species-poor clusters.We concluded that tree species identity and site conditions were more important factors determining the soil microbial community structure than tree species diversity per se.     

Functional and molecular responses of soil microbial communities under differing soil management practices

The effects of soil management on some microbiological properties and soil bacterial community structure were evaluated. Two field sites with the same soil type, located on the same geographic area adjacent to one other, have received different soil management practices and cultivation. One site has been subjected for 20 years to intensive horticulture under conventional tillage and irrigation with low quality salt-rich water; the second field site has been uncultivated for a long period and was turned to organic farming practices over the last 5 years and is currently cultivated with fruit orchard. Total bacterial counts, microbial ATP, microbial community metabolic (BIOLOG^®) profiles, and DNA fingerprinting by PCR-DGGE were determined. Two-way ANOVA revealed that total bacterial counts were not significantly (P>0.3) affected by the two different management practices; ATP content was consistently and significantly (P<0.001) lower in salt-water irrigated soil than in organic soil at the three sampling times. The cluster analysis of community level physiological profiles indicated that microbial communities were much more uniform in organic soil than in irrigated one, suggesting that salt-water irrigation could have affected the size of the microbial population, its metabolic activities, as well as its composition. Molecular patterns fitted the BIOLOG^® profile diversity. In particular, at any sampling time, PCR-DGGE patterns of bacterial DNA, extracted by an indirect method, significantly discriminated irrigated from organic soil samples. The PCR-DGGE patterns of total soil DNA, extracted by a direct method, showed a moderate to significant variation among irrigated and organic soil samples. Biochemical, microbiological and molecular data contributed to evidence a significantly different response of indigenous microflora to soil management by using saline water or organic farming.     

Interactions between plant species and mycorrhizal colonization on the bacterial community composition in the rhizosphere

This study assessed the effect of mycorrhizal colonization by Glomus intraradices (Gi) and G. versiforme (Gv) on the bacterial community composition in the rhizosphere of canola, clover and two tomato genotypes (wild type (76R) and its mutant with reduced mycorrhizal colonization (rmc)). Additionally, the effect of light intensity on the rhizosphere bacterial community composition of the tomato genotypes was studied. The bacterial community composition was assessed by denaturing gradient gel electrophoresis (DGGE). In canola, which is considered to be a non-mycorrhizal species, inoculation with Gi increased the shoot dw compared to Gv and the non-mycorrhizal control plants and also induced changes in the bacterial community composition in the rhizosphere. These fungal effects were observed although less than 8% of the root length of canola was colonized. On the other hand, about 50% of the root length of clover was colonized and inoculation with Gv resulted in a higher shoot dw compared to Gi or the control plants but the rhizosphere bacterial community composition was not affected by inoculation. Plant growth, mycorrhizal colonization and bacterial community composition of the two tomato genotypes were affected by a complex interaction between tomato genotype, AM fungal species and light intensity. Low light intensity (photosynthetic photon flux 200–250 μmol m⁻² s⁻¹) increased the shoot–root ratio in both genotypes and reduced colonization in the wild type. The differences in bacterial community composition between the two genotypes were more pronounced at low than at high light intensity (550–650 μmol m⁻² s⁻¹).     

Structural and functional diversity of bacterial community in soil treated with the herbicide napropamide estimated by the DGGE,CLPP and r/K-strategy approaches

Napropamide is one of the most commonly used herbicide in agricultural practice and can exhibit toxic effect to soil microorganisms. Therefore, the main objective of this study was to examine the genetic and functional diversity of microbial communities in soil treated with napropamide at field rate (FR, 2.25 mg kg⁻¹ of soil) and 10 times the FR (10 × FR, 22.5 mg kg⁻¹ of soil) by the denaturing gradient gel electrophoresis (DGGE) and the community level physiological profile (CLPP) methods. In addition, the r/K-strategy approach was used to evaluate the effect of this herbicide on the community structure of the culturable soil bacteria. DGGE patterns revealed that napropamide affected the structure of microbial community; however, the richness (S) and genetic diversity (H) values indicated that the FR dosage of napropamide experienced non-significant changes. In turn, the 10 × FR dosage of herbicide caused significant changes in the S and H values of dominant soil bacteria. DGGE profiles suggest an evolution of bacteria capable of degrading napropamide among indigenous microflora. Analysis of the CLPPs indicated that the catabolic activity of microbial community expressed as AWCD (average well-color development) was temporary positively affected after napropamide application and resulted in an increase of the substrate richness (S_R) as well as functional biodiversity (H) values. Analysis of the bacterial growth strategy revealed that napropamide affected the r- or K-type bacterial classes (ecotypes). In treated-soil samples K-strategists dominated the population, as indicated by the decreased ecophysiological (EP) index. Napropamide significantly affected the physiological state of culturable bacteria and caused a reduction in the rate of colony formation as well as a prolonged time of growth rate. Obtained results indicate that application of napropamide may poses a potential risk for soil functioning.     

Soil microbial community response to land-management and depth,related to the degradation of organic matter in English wetlands: Implications for the in situ preservation of archaeological remains

Wetlands are important habitats not only for their unique ecological value but also because they contain organic material that is fundamental to our understanding of precedent landscape and human past. This study compares the effects of two different land-management regimes on metabolic diversity and bacterial community structure with depth in order to relate them to the process of organic matter degradation and the potential for preservation in situ of organic archaeological artefacts in wetland soils. Soil cores were collected at five depths down to 100 cm from two wetlands sites in England. Environmental variables were monitored and the metabolic capabilities of the microbial community were studied using Biolog Ecoplates^®. DNA was extracted from soil, and the bacterial community structure was examined by polymerase chain reaction followed by denaturing gradient gel electrophoresis (PCR-DGGE). To determine compositional changes in the bacterial community with depth, information about specific groups of bacteria at the site with higher water table (Hatfield Moor) was obtained by cloning and sequencing of 16S rRNA genes. Biolog and DGGE analyses showed depth variation and between-site variation. Carbon substrate utilization and bacterial diversity decreased with increasing depth. The wetland soil under an arable regime in which the water levels were kept elevated, showed higher metabolic capability and bacterial richness when compared with the soil under pasture and subjected to long-standing drainage. Cloning and sequencing showed that Proteobacteria and Acidobacteria were the predominant taxa within the soil profile, but there was a clear shift in bacterial community composition with increasing depth as several taxonomic groups (δ-Proteobacteria and Spirochaetes) were only detectable at 50 cm depth. Because the site with a high and stable water table presented higher metabolic activity and bacterial diversity, it may be that saturated conditions and a high water table are not sufficient to guarantee the preservation in situ of organic material such as archaeological artefacts.     

Pyrosequencing and mid-infrared spectroscopy reveal distinct aggregate stratification of soil bacterial communities and organic matter composition

This study integrated physical, chemical, and molecular techniques to assess relationships between soil bacterial community structures and the quantity and quality of soil organic carbon (SOC) at the soil microenvironment scale (e.g., within different aggregate size-fractions). To accomplish this goal, soil samples (0–5 cm) were collected from the Texas High Plains region under a variety of dryland and irrigated cropping systems. The soil was separated into macroaggregates, microaggregates, and silt + clay fractions that were analyzed for (1) bacterial diversity via pyrosequencing of the 16s rRNA gene and (2) SOC quantity and quality using a combustion method and mid-infrared diffuse reflectance spectroscopy (mid-IR), respectively. Results from pyrosequencing showed that each soil microenvironment supported a distinct bacterial community. Similarly, mid-IR data revealed distinct spectral features indicating that these fractions were also distinguished by organic and mineral composition. Macroaggregates showed relatively high abundance of Actinobacteria (excluding order Rubrobacteriales) and α-Proteobacteria and contained the most SOC. Microaggregates showed high relative abundance of Rubrobacteriales and the least amount of SOC. Predominance within the soil microenvironment and correlations along the mid-IR spectra were different between members of the order Rubrobacteriales compared with all other members of the Actinobacteria phyla, suggesting they have different ecological niches. Mid-IR results revealed microaggregates had greater absorbance in the 1370–1450 cm^?1 region for phenolic and alkyl groups (possibly recalcitrant C). Silt + clay fractions were distinguished by Gemmatimonadetes and OP10 phyla, which positively correlated with spectral absorption in the1250–1150 cm^?1 range (indicating both degradable and recalcitrant C forms). In contrast to general diversity index measurements, distributions of the more rare bacterial phyla (phyla representing <6% of the identified population) were more important for differentiating between communities in soil microenvironments. To our knowledge, this is the first study to investigate soil bacterial communities among soil aggregates using pyrosequenging and to associate these communities to specific soil C chemistries as indicated by mid-IR absorbance.     

Comparison of root system architecture and rhizosphere microbial communities of Balsas teosinte and domesticated corn cultivars

The progenitor of maize is Balsas teosinte (Zea mays subsp. parviglumis) which grows as a wild plant in the valley of the Balsas river in Mexico. Domestication, primarily targeting above-ground traits, has led to substantial changes in the plant's morphology and modern maize cultivars poorly resemble their wild ancestor. We examined the hypotheses that Balsas teosinte (accession PI 384071) has a) a different root system architecture and b) a structurally and functionally different rhizosphere microbial community than domesticated cultivars sweet corn (Zea mays subsp. mays accession PI 494083) and popping corn (Zea mays subsp. mays accession PI 542713). In a greenhouse experiment, five plants from each corn variety were grown in individual pots containing a Maury silt loam – perlite (2:1) mixture and grown to the V8 growth stage at which rhizosphere bacterial and fungal community structure was assessed using terminal restriction fragment length polymorphism and fatty acid methyl ester analysis. Functional characteristics of the rhizosphere were assayed by examining the potential activity of seven extracellular enzymes involved in carbon, nitrogen and phosphorus cycling. Root system architecture was characterized by root scans of sand grown plants at the V5 growth stage. Compared to the control the sweet corn rhizosphere had different bacterial and fungal community structure, decreased fungal diversity and increased bacterial abundance. Teosinte caused a significant change in the rhizosphere bacterial and fungal community structure and increased bacterial abundance, but no significant decrease in bacterial or fungal diversity where the former was found to be significantly greater than in the sweet corn rhizosphere. Popping corn did not trigger significant changes in the bacterial or fungal diversity and bacterial abundance in the soil. The individual popping corn plants changed the bacterial and fungal communities in different directions and the overall effect on community structure was significant, but small. Of the enzymes analyzed, potential N-acetylglucosaminidase (NAG) activity was found to contributed most to the differentiation of teosinte rhizosphere samples from the other corn varieties. The teosinte root system had proportionally more very fine (diameter < 0.03 mm) roots than popping corn and sweet corn and it developed the highest root to shoot dry weight ratio, followed by popping corn. Sweet corn had significantly lower average root diameter than popping corn and teosinte and grew proportionally the least below-ground dry mass. The results allude to functional and structural differences in the rhizosphere microbial communities of the corn varieties that, with additional research, could lead to useful discoveries on how corn domestication has altered rhizosphere processes and how plant genotype influences nutrient cycling.     

Changes of arbuscular mycorrhizal traits and community structure with respect to soil salinity in a coastal reclamation land

A comprehensive knowledge on the relationship between soil salinity and arbuscular mycorrhizal fungi (AMF) is vital for a deeper understanding of ecosystem functioning under salt stress conditions. The objective of this study was to determine the effects of soil salinity on AMF root colonization, spore count, glomalin related soil protein (GRSP) and community structure in Saemangeum reclaimed land, South Korea. Soil samples were collected and grouped into five distinct salt classes based on the electrical conductivity of soil saturation extracts (EC_se). Mycorrhizal root colonization, spore count and GRSP were measured under different salinity levels. AMF community structure was studied through three complementary methods; spore morphology, terminal restriction fragment length polymorphism (T-RFLP) and denaturing gradient gel electrophoresis (DGGE). Results revealed that root colonization (P < 0.01), spore count (P < 0.01) and GRSP (P < 0.01) were affected negatively by soil salinity. Spore morphology and T-RFLP data showed predominance of AMF genus Glomus in Saemangeum reclaimed land. T-RFLP and DGGE analysis revealed significant changes in diversity indices between non (EC_se < 2 dS/m) and extremely (EC_se > 16 dS/m) saline soil and confirmed dominance of Glomus caledonium only in soils with EC_se < 8 dS/m. However, ribotypes of Glomus mosseae and Glomus proliferum were ubiquitous in all salt classes. Combining spore morphology, T-RFLP and DGGE analysis, we could show a pronounced effect in AMF community across salt classes. The result of this study improve our understanding on AMF activity and dominant species present in different salt classes and will substantially expand our knowledge on AMF diversity in reclaimed lands.     

Bacillus amyloliquefaciens BNM122, a potential microbial biocontrol agent applied on soybean seeds,causes a minor impact on rhizosphere and soil microbial communities

The increase in soybean productivity has contributed to a greater use of agrochemicals, which cause major problems, such as soil and water pollution and reduction of biodiversity, and have a negative impact on non-target species. The development of microbial biocontrol agents for soybean diseases can help to reduce pesticide abuse. Bacillus amyloliquefaciens BNM122 is a potential microbial biocontrol agent able to control the damping-off caused by Rhizoctonia solani when inoculated in soybean seeds, both in a plant growth chamber and in a greenhouse. In this study, we report the effect of soybean seed treatments with strain BNM122 or with two fungicides (thiram and carbendazim) on the structure and function of the bacterial community that colonizes the soybean rhizosphere. Also, soybean root nodulation by Bradyrhizobium japonicum, mycorrhization by arbuscular mycorrhizal fungi and plant growth were evaluated. We used the r- and K-strategist concept to evaluate the ecophysiological structure of the culturable bacterial community, community-level physiological profiles (CLPP) in Biolog? EcoPlates to study bacterial functionality, and the patterns of 16S RNA genes amplified by PCR and separated by denaturing gradient gel electrophoresis (PCR–DGGE) to assess the genetic structure of the bacterial community. Neither the ecophysiological structure nor the physiological profiles of the soybean rhizosphere bacterial community showed important changes after seed inoculation with strain BNM122. On the contrary, seed treatment with fungicides increased the proportions of r-strategists and altered the metabolic profiles of the rhizosphere culturable bacterial community. The genetic structure of the rhizosphere bacterial community did not show perceptible changes between treated and non-treated seeds. Regarding the bacterial and fungal symbioses, seed treatments did not affect soybean nodulation, whereas soybean mycorrhization significantly decreased (P < 0.05) in plants obtained from seeds treated with strain BNM122 or with the fungicides. However, a higher negative effect was observed in plants which seeds were treated with the fungicides. Plant growth was not affected by seed treatments.It can be concluded that soybean seed treatment with B. amyloliquefaciens BNM122 had a lesser effect on soil microbial community than that with the fungicides, and that these differences may be attributed to the less environmental persistence and toxic effects of the strain, which deserve further studies in order to develop commercial formulations.     

Characterization of copper-resistant bacterial community in rhizosphere of highly copper-contaminated soil

The characteristics of copper (Cu)-resistant bacterial communities in a rhizosphere and a non-rhizosphere of the ditch reed, Phragmites, in a highly Cu-contaminated area near a copper mine were investigated. The total Cu concentration was 720 μg·g^–1 in the rhizosphere soil and 5 680 μg·g^–1 in the non-rhizosphere soil. In the rhizosphere, the multiplication of bacteria, particularly non-resistant bacteria, was promoted as compared with the non-rhizosphere. The properties of the bacterial community in the rhizosphere were quite different from those in the non-rhizosphere in terms of Cu sorption ability, growth rate and exopolymer production. Both the Cu-resistant bacteria and non-resistant bacteria in the rhizosphere grew more rapidly in media than those in the non-rhizosphere. For almost all the isolated Cu-resistant bacteria, exopolymer production was prompted by Cu stimuli especially for the isolates from the rhizosphere. The adverse effect of Cu on the growth rate was found to be small for the Cu-resistant bacteria producing exopolymers in a large quantity, suggesting the involvement of exopolymers in the detoxification of Cu. The role of Cu resistance in the bacterial chemotactic migration in the Cu-contaminated soils was not evident.