Microbial community development in the rhizosphere of apple trees at a replant disease site

Apple replant disease (ARD) is a complex syndrome that affects young trees in replanted orchard sites causing necrotic lesions on feeder roots, stunted tree growth and reduced cumulative yields. Use of ARD-tolerant rootstocks is an emerging control strategy. We studied the bacterial, fungal, and oomycetes populations in the rhizosphere of five rootstock cultivars (M.7, M.26, G.16, G.30 and CG.6210) planted into the old tree row or grass lanes of a previous orchard in Ithaca, NY, to better understand the role of rhizosphere microbial communities in the prevalence and control of ARD. The possible involvement of antagonistic Pseudomonas species, Pythium spp., Phytophthora spp. and rhizosphere cyanide concentrations in ARD were also examined. The rootstocks M.7, M.26 and G.16 were susceptible to ARD, while G.30 and CG.6210 were more tolerant. Tree growth on the rootstocks M.7, M.26 and G.16 was reduced by 10% when planted in the old tree rows, but this did not significantly reduce yields in the first fruiting year. The susceptible rootstocks, M.7 and M.26, supported higher densities of culturable rhizosphere fungi and bacteria than G.16, G.30 and CG.6210. Over 2 years, microbial densities were highest in July, lower in May and lowest in September. The composition of bacterial and fungal communities in the rhizosphere was highly variable and changed over seasons and years, as assessed by terminal restriction fragment length polymorphism (T-RFLP) analyses. Initial differences in fungal rhizosphere communities between the two planting positions converged 2 years after the trees were replanted. In contrast, the bacterial rhizosphere community composition still differed significantly between the two planting positions 3 years after the orchard was replanted. The bacterial and fungal rhizosphere community compositions of susceptible rootstocks, M.7 and M.26, differed from those of the tolerant rootstocks, G.30 and CG.6210; G.16, differed from all the other rootstocks. The observed effects of rootstocks, planting positions and time on microbial community composition were small relative to the high variability observed overall. Pythium spp. and Phytophthora spp. infestations were high and similar for all rootstocks and planting positions. Neither potentially antagonistic Pseudomonas nor rhizosphere cyanide concentrations appeared to be involved in the ARD-complex at the studied site. Avoiding replanting into the old tree rows coupled with use of tolerant rootstocks appear to be the best strategies for reducing ARD in replanted orchards. Changes in rhizosphere microbial communities are among the many factors that contribute to improved tree growth when these management strategies are used.     

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.     

Shrub-interspace dynamics alter relationships between microbial community composition and belowground ecosystem characteristics

In desert ecosystems, belowground characteristics are influenced chiefly by the formation and persistence of “shrub-islands of fertility” in contrast to barren plant interspaces. If soil microbial communities are exclusively compared between these two biogeochemically distinct soil types, the impact of characteristics altered by shrub species, especially soil C and N, are likely to be overemphasized and overshadow the role of other characteristics in structuring microbial composition. To determine how belowground characteristics influence microbial community composition, and if the relative importance of these characteristics shifts across the landscape (i.e., between and within shrub and interspace soils), changes in microbial communities across a 3000-year cold desert chronosequence were related to 27 belowground characteristics in surface and subsurface soils. When shrub and interspace communities in surface and subsurface soils were combined across the entire chronosequence, communities differed and were primarily influenced by soil C, NO₃⁻ concentrations, bulk density, pH, and root presence. Within shrub soils, microbial communities were shrub species-specific, especially in surface soils, highlighting differences in soil characteristics created by specific shrub species and/or similarity in stresses structuring shrub species and microbial communities alike. Microbial communities in shrub soils were not influenced by soil C, but by NO₃⁻ and NH₄⁺ concentrations, pH, and silt in surface soils; and Cl, P, soil N, and NO₃⁻ concentrations in subsurface soils. Interspace soil communities were distinct across the chronosequence at both depths and were strongly influenced by dune development. Interspace communities were primarily associated with soil stresses (i.e., high B and Cl concentrations), which decreased with dune development. The distribution of Gram-positive bacteria, Actinobacteria, and fungi highlighted community differences between and within shrub and interspace soils, while Gram-negative bacteria were common in all soils across the chronosequence. Of the 27 belowground characteristics investigated, 13 separated shrub from interspace communities, and of those, only five emerged as factors influencing community composition within shrub and interspace soils. As dunes develop across this cold desert chronosequence, microbial community composition was not regulated primarily by soil C, but by N and P availability and soil stresses in shrub soils, and exclusively by soil stresses in interspace soils.     

Survival of Escherichia coli in soil with modified microbial community composition

Understanding the survival and persistence of Escherichia coli in soil with different microbial composition is essential for the accuracy of water quality assessment and microbial source tracking. This microcosm experiment was conducted to investigate the survival pattern of three E. coli strains (originated from soil, dog feces and human feces, separately) in soil with modified microbial community composition. Bile salt No. 3 (BS3) of progressively increased density (0.05%, 0.15%, 0.30% and 0.50%) was added into sandy loam soils and incubated for 90 days. Laboratory cultured E. coli were then inoculated into soil and incubated for another 150 days to monitor their survival pattern. Change of bacterial community diversity by BS3 was detected by both cultivation based and cultivation independent (PCR-Denaturing Gradient Gel Electrophoresis) methods. In general, progressively increased BS3 concentration resulted in decreased CFU counts both at 10 days and 90 days incubation. DGGE analysis indicated only a slight change in bacterial community composition at 10 days but a significant change at 90 days. Cluster analysis suggested that BS3 treatment grouped separately from controls. Survival of E. coli in soil was significantly influenced by the complexity of the microbial community, as die-off rate of E. coli progressively declined with the reduction of microbial community diversity. Differential survival of E. coli under different soil microbial stress highlights the importance of incorporating biotic factors in predictive models for water quality management and microbial source tracking study.     

Litter type, but not plant cover, regulates initial litter decomposition and fungal community structure in a recolonising cutover peatland

Cutover peatlands are often rapidly colonised by pioneer plant species, which have the potential to affect key ecosystem processes such as carbon (C) turnover. The aim of this study was to investigate how plant cover and litter type affect fungal community structure and litter decomposition in a cutover peatland. Intact cores containing Eriophorum vaginatum, Eriophorum angustifolium, Calluna vulgaris and bare soil were removed and a mesh bag with litter from only one of each of these species or fragments of the moss Sphagnum auriculatum was added to each core in a factorial design. The presence or absence of live plants, regardless of the species, had no effect on mass loss, C, nitrogen (N) or phosphorus (P) concentrations of the litter following 12 months of incubation. However, there was a very strong effect of litter type on mass loss and concentrations of C, N and P between most combinations of litter. Similarly, plant species did not affect fungal community structure but litter type had a strong effect, with significant differences between most pairs of litter types. The data suggest that labile C inputs via rhizodeposition from a range of plant functional types that have colonised cutover bogs for 10-15 years have little direct effect on nutrient turnover from plant litter and in shaping litter fungal community structure. In contrast, the chemistry of the litter they produce has much stronger and varied effects on decomposition and fungal community composition. Thus it appears that there is distinct niche differentiation between the fungal communities involved in turnover of litter versus rhizodeposits in the early phases of plant succession on regenerating cutover peatlands.     

Elevated CO2 differentially alters belowground plant and soil microbial community structure in reed canary grass-invaded experimental wetlands

Several recent studies have indicated that an enriched atmosphere of carbon dioxide (CO₂) could exacerbate the intensity of plant invasions within natural ecosystems, but little is known of how rising CO₂ impacts the belowground characteristics of these invaded systems. In this study, we examined the effects of elevated CO₂ and nitrogen (N) inputs on plant and soil microbial community characteristics of plant communities invaded by reed canary grass, Phalaris arundinacea L. We grew the invasive grass under two levels of invasion: the invader was either dominant (high invasion) at >90% plant cover or sub-dominant (low invasion) at <50% plant cover. Experimental wetland communities were grown for four months in greenhouses that received either 600 or 365 μl l⁻¹ (ambient) CO₂. Within each of three replicate rooms per CO₂ treatment, the plant communities were grown under high (30 mg l⁻¹) or low (5 mg l⁻¹) N. In contrast to what is often predicted under N limitation, we found that elevated CO₂ increased native graminoid biomass at low N, but not at high N. The aboveground biomass of reed canary grass did not respond to elevated CO₂, despite it being a fast-growing C3 species. Although elevated CO₂ had no impact on the plant biomass of heavily invaded communities, the relative abundance of several soil microbial indicators increased. In contrast, the moderately invaded plant communities displayed increased total root biomass under elevated CO_2, while little impact occurred on the relative abundance of soil microbial indicators. Principal components analysis indicated that overall soil microbial community structure was distinct by CO₂ level for the varying N and invasion treatments. This study demonstrates that even when elevated CO₂ does not have visible effects on aboveground plant biomass, it can have large impacts belowground.     

Methane oxidation activity and bacterial community composition in a simulated landfill cover soil is influenced by the growth of Chenopodium album L.

Oxygen availability in landfill cover soil is a major limitation to the growth and activity of methanotrophs as methane oxidation is an aerobic microbial process. Plants tolerant to high concentrations of landfill gas (LFG) may play an important role in improving methane oxidation within landfill cover soil and reducing emission of methane, a greenhouse gas, from it. In this study, the effect of an LFG tolerant plant Chenopodium album L. on methane oxidation activity (MOA) and bacterial community composition in landfill cover soil was investigated. Soil samples from four simulated lysimeters with and without LFG and plant vegetation were taken at 4 stages during the plant's development cycle. Results showed that the total number of culturable bacteria in soil could be significantly increased (P < 0.05) by the growth of C. album. The total number of methanotrophs and MOA in soils with LFG was significantly higher (P < 0.05) than in soils without LFG on sampling days 90, 150 and 210. The total number of methanotrophs and MOA in lysimeters with LFG added increased in the presence of C. album when the plant entered the seed setting stage. Polymerase chain reaction and denaturing gradient gel electrophoresis (PCR-DGGE) gel patterns of 16S rDNA gene fragment and band sequencing analyses showed apparent differences in soil bacterial communities in the presence of LFG and plant vegetation. Members of the genus Methylosarcina were found to be the active and dominant methanotrophs in rhizosphere soil of C. album with LFG, while Methylococcus, Methylocystis, and Methylosinus were the primary methanotroph genera in LFG soil without C. album. Thus, C. album appears to select for specific methanotrophic bacteria in the presence of LFG. Soil MOA and microbial diversity can also be significantly affected by the presence of this plant.     

Microbial community structure and function in a soil contaminated by heavy metals: effects of plant growth and different amendments

We studied the effects of in situ remediation of a heavy metal (HM) contaminated soil on some soil chemical properties, microbial function and microbial structural diversity after 18 months. The experiment was carried out at semifield scale in containers filled with HM contaminated soil from the Aznalcóllar mine accident (Southern Spain, 1998). The remediation measures consisted of the application of different amendments and/or establishment of a plant cover (Agrostis stolonifera L.). Seven treatments were established: four organic treatments (municipal waste compost (MWC), biosolid compost (BC), leonardite (LEO) and litter (LIT)), one inorganic treatment (sugar beet lime (SL)) and two controls (control with plant cover (CTRP) and control without plant cover (CTR)). Several soil chemical (pH, soluble HM, total organic C (TOC), water-soluble C (WSC) and available-P) and biochemical properties (microbial biomass C (MBC), MBC/TOC ratio and enzyme activities) were determined. Microbial community structure was studied by means of ARDRA (amplified ribosomal DNA restriction analysis). The SL, MWC and BC treatments were the most efficient to raise soil pH and decrease soluble HM concentrations. Total organic C was increased in the organic treatments by 2 to 4-fold, whereas water-soluble C was statistically similar in the CTRP, SL and the organic treatments, probably due to the presence of a root system in all these treatments. Available-P was also increased in the BC, SL and MWC treatments due to the higher P content of the amendments applied in these treatments. Soil microbial function was generally enhanced in the amended and CTRP treatments. The MWC, BC and SL treatments were particularly efficient to increase microbial biomass C, the MBC/TOC ratio and the dehydrogenase and aryl-sulphatase enzyme activities. These results could be attributed to the amelioration of some of the soil chemical properties: increase in soil pH and water-soluble C and decrease of HM soluble concentrations. ARDRA analyses showed changes in structural diversity in both the bacterial and fungal community under the different treatments. Fingerprinting patterns of the 16S rDNA obtained with Hinf-I and of the 18S rDNA with Hpa-II revealed higher similarity percentages among samples from the same treatment compared with samples from the other treatments. In addition, a higher similarity was found between samples from all treatments under the Agrostis influence. The use of certain amendments and/or a plant cover is important for in situ remediation of HM contaminated soils, since these practices can affect soil chemical properties, as well as the microbial community function and structure.