Fine-Scale Horizontal and Vertical Micro-distribution Patterns of Testate Amoebae Along a Narrow Fen/Bog Gradient

The ecology of peatland testate amoebae is well studied along broad gradient from very wet (pool) to dry (hummock) micro-sites where testate amoebae are often found to respond primarily to the depth to water table (DWT). Much less is known on their responses to finer-scale gradients, and nothing is known of their possible response to phenolic compounds, which play a key role in carbon storage in peatlands. We studied the vertical (0–3, 3–6, and 6–9 cm sampling depths) micro-distribution patterns of testate amoebae in the same microhabitat (Sphagnum fallax lawn) along a narrow ecological gradient between a poor fen with an almost flat and homogeneous Sphagnum carpet (fen) and a “young bog” (bog) with more marked micro-topography and mosaic of poor fen and bog vegetation. We analyzed the relationships between the testate amoeba data and three sets of variables (1) “chemical” (pH, Eh potential, and conductivity), (2) “physical” (water temperature, altitude, i.e., Sphagnum mat micro-topography, and DWT), and (3) phenolic compounds in/from Sphagnum (water-soluble and primarily bound phenolics) as well as the habitat (fen/bog) and the sampling depth. Testate amoeba Shannon H′ diversity, equitability J of communities, and total density peaked in lower parts of Sphagnum, but the patterns differed between the fen and bog micro-sites. Redundancy analyses revealed that testate amoeba communities differed significantly in relation to Eh, conductivity, water temperature, altitude, water-soluble phenolics, habitat, and sampling depth, but not to DWT, pH, or primarily bound phenolics. The sensitivity of testate amoebae to weak environmental gradients makes them particularly good integrators of micro-environmental variations and has implications for their use in paleoecology and environmental monitoring. The correlation between testate amoeba communities and the concentration of water-soluble phenolic suggests direct (e.g., physiological) and/or indirect (e.g., through impact on prey organisms) effects on testate amoebae, which requires further research.     

Sphagnum physiological responses to elevated temperature,nitrogen, CO2 and low moisture in laboratory and in situ microhabitats: a review

Aquatic Ecology - Sphagnum mosses are considered peatland engineers because of their ability to create conditions inducing carbon accumulation. Here, we report on a review of the effects of four...     

Above‐ and belowground linkages in Sphagnum peatland: climate warming affects plant‐microbial interactions

Peatlands contain approximately one third of all soil organic carbon (SOC). Warming can alter above‐ and belowground linkages that regulate soil organic carbon dynamics and C‐balance in peatlands. Here we examine the multiyear impact of in situ experimental warming on the microbial food web, vegetation, and their feedbacks with soil chemistry. We provide evidence of both positive and negative impacts of warming on specific microbial functional groups, leading to destabilization of the microbial food web. We observed a strong reduction (70%) in the biomass of top‐predators (testate amoebae) in warmed plots. Such a loss caused a shortening of microbial food chains, which in turn stimulated microbial activity, leading to slight increases in levels of nutrients and labile C in water. We further show that warming altered the regulatory role of Sphagnum‐polyphenols on microbial community structure with a potential inhibition of top predators. In addition, warming caused a decrease in Sphagnum cover and an increase in vascular plant cover. Using structural equation modelling, we show that changes in the microbial food web affected the relationships between plants, soil water chemistry, and microbial communities. These results suggest that warming will destabilize C and nutrient recycling of peatlands via changes in above‐ and belowground linkages, and therefore, the microbial food web associated with mosses will feedback positively to global warming by destabilizing the carbon cycle. This study confirms that microbial food webs thus constitute a key element in the functioning of peatland ecosystems. Their study can help understand how mosses, as ecosystem engineers, tightly regulate biogeochemical cycling and climate feedback in peatlands     

Effect of a temperature gradient on Sphagnum fallax and its associated living microbial communities: a study under controlled conditions

Microbial communities living in Sphagnum are known to constitute early indicators of ecosystem disturbances, but little is known about their response (including their trophic relationships) to climate change. A microcosm experiment was designed to test the effects of a temperature gradient (15, 20, and 25°C) on microbial communities including different trophic groups (primary producers, decomposers, and unicellular predators) in Sphagnum segments (0-3 cm and 3-6 cm of the capitulum). Relationships between microbial communities and abiotic factors (pH, conductivity, temperature, and polyphenols) were also studied. The density and the biomass of testate amoebae in Sphagnum upper segments increased and their community structure changed in heated treatments. The biomass of testate amoebae was linked to the biomass of bacteria and to the total biomass of other groups added and, thus, suggests that indirect effects on the food web structure occurred. Redundancy analysis revealed that microbial assemblages differed strongly in Sphagnum upper segments along a temperature gradient in relation to abiotic factors. The sensitivity of these assemblages made them interesting indicators of climate change. Phenolic compounds represented an important explicative factor in microbial assemblages and outlined the potential direct and (or) indirect effects of phenolics on microbial communities.     

Uptake and translocation of phytochemical 2-benzoxazolinone (BOA) in radish seeds and seedlings

The molecular aspects of phytochemical interactions between plants, especially the process of phytochemical translocation by the target plant, remain challenging for those studying allelopathy. 2-Benzoxazolinone (BOA) is a natural chemical produced by rye (Secale cereale) and is known to have phytotoxic effects on weed seeds and seedlings. The translocation of BOA into target plants has been poorly investigated. Therefore, the total absorption of [ring U 14C] BOA was estimated by oxidizing whole seedlings of Raphanus sativus cv. for 8 days and quantifying the radioactivity. Non-radiolabelled BOA in seedlings was also estimated by HPLC. BOA applied at 10(-3) M was readily taken up by germinated radish at a rate of 1556 nmol g(-1) FW. At these same concentrations, BOA reduced radish germination by 50% and caused a delay in radicle elongation. Exogenous BOA was responsible for the observed germination inhibition. At a concentration of 10(-5) M, BOA was taken up by germinated seeds (31 nmol g(-1) FW), but this quantity did not affect radish germination. Labelled BOA was not mineralized in the culture medium during seedling growth as no 14CO2 was recovered. Both 10(-3) and 10(-5) M BOA were translocated into radish organs, mainly into roots and cotyledons. These organs were then identified as potential physiological target sites. Cotyledons remained the target sink (44% of the total radioactivity). The kinetics of BOA uptake at 10(-3) and 10(-5) M in radish seedlings was identical: BOA accumulation was proportional to its initial concentration. A comparison between radioactivity and HPLC quantification for 10(-3) M BOA indicated that BOA (along with some metabolites) could effectively be recovered in radish organs using chromatography.     

Benzoxazolin-2(3H)-one (BOA) induced changes in leaf water relations,photosynthesis and carbon isotope discrimination in Lactuca sativa

The effects are reported here of Benzoxazolin-2(3H)-one (BOA), an allelopathic compound, on plant water relations, growth, components of chlorophyll fluorescence, and carbon isotope discrimination in lettuce (Lactuca sativa L.). Lettuce seedlings were grown in 1:1 Hoagland solution in perlite culture medium in environmentally controlled glasshouse. After 30 days, BOA was applied at concentration of 0.1, 0.5, 1.0 and 1.5 mM and distilled water (control). BOA, in the range (0.1–1.5 mM), decreased the shoot length, root length, leaf and root fresh weight. Within this concentration range, BOA significantly reduced relative water content while leaf osmotic potential remained unaltered. Stress response of lettuce was evaluated on the basis of six days of treatment with 1.5 mM BOA by analyzing several chlorophyll fluorescence parameters determined under dark-adapted and steady state conditions. There was no change in initial fluorescence (F_o) in response to BOA treatment while maximum chlorophyll fluorescence (F_m) was significantly reduced. BOA treatment significantly reduced variable fluorescence (F_v) on first, second, third, fourth, fifth and sixth day. Quantum efficiency of open PSII reaction centers (F_v/F_m) in the dark-adapted state was significantly reduced in response to BOA treatment. Quantum yield of photosystem II (ΦPSII) electron transport was significantly reduced because of decrease in the efficiency of excitation energy trapping of PSII reaction centers. Maximum fluorescence in light-adapted leaves (F′_m) was significantly decreased but there was no change in initial fluorescence in light-adapted state (F′_o) in response to 1.5 mM BOA treatment. BOA application significantly reduced photochemical fluorescence quenching (qP) indicating that the balance between excitation rate and electron transfer rate has changed leading to a more reduced state of PSII reaction centers. Non photochemical quenching (NPQ) was also significantly reduced by BOA treatment on third, fourth and fifth day. BOA had dominant effect on C isotope ratios (δ¹³C) that was significantly less negative (?26.93) at 1.0 mM concentration as compared to control (?27.61). Carbon isotope discrimination (Δ¹³C) values were significantly less (19.45) as compared to control (20.17) at 1.0 mM. BOA also affect ratio of intercellular to air CO₂ concentration (ci/ca) that was significantly less (0.66) as compared to control (0.69) when treated with 1.0 mM BOA. Protein content of lettuce leaf tissue decreased under BOA treatment at 1.5 mM concentration as compared to control.     

Two biochemical forms of phenanthrene recovered in grassland plants (Lolium perenne L. and Trifolium pratense L.) grown in 3 spiked soils

Plant storage is a key component in the global cycling of persistent polycyclic aromatic hydrocarbons (PAHs) and constitutes a crucial step to understand environmental fate of such pollutants. This work aimed at quantifying biochemical forms of phenanthrene (PHE) stored in plants and determining the main parameter between the plant species and the soil characteristics involved in the PHE bioaccumulation. An experiment was conducted in a growth chamber to study the storage of PHE in ryegrass and clover in three distinct artificially contaminated soils (1,000 ??g g^?1 DW). A preliminary experiment was designed to develop an extraction method which distinguished easily extractable PHE (free-PHE) from less extractable PHE (bound-PHE). PHE was found to be mainly recovered in plant roots (around 90% of the total PHE recovered in plants) in its free form. PHE was transported upward from the roots into shoots. PHE recovered in shoots (around 1 ??g g^?1) was divided into its free (60%) and bound (40%) forms except for one treatment. Observed differences of root PHE storage were clearly related with the PHE dissipation in soil and then with the plant species. This work outlined the importance of quantifying the bound and free PAHs in plants in further researches.     

Atmospheric phenanthrene transfer and effects on two grassland species and their root symbionts: A microcosm study

The objective of this work was to determine the transfer of phenanthrene (PHE) from air to grassland plants and soil compartments and its effects on the plant growth and symbiotic root microorganisms (arbuscular mycorrhizal fungi and Rhizobium nodules). The experimental procedure exposed Trifolium pratense L. or Lolium perenne L. to atmospheric PHE pollution (150 μg m⁻³) over the course of one month. PHE was transferred from the air to the leaves and to the soil surfaces. In leaves, PHE was mostly absorbed in the inner leaf tissues, representing 92% and 73% of the total PHE amount quantified in leaves, respectively for clover and ryegrass. In soils, most of PHE contamination was recovered in the top layer (0-1 cm) and did not readily diffuse into the deep layer (1-10 cm). The highest PHE concentration recovered in deep roots (1.8 and 4.5 μg g⁻¹ dry weight (DW), respectively for clover and ryegrass) related to the lowest PHE concentration recovered in its associated soil suggested a PHE translocation from shoots to roots within the two plant species. The large PHE amount quantified in clover shoots (124 μg g⁻¹ DW) induced a significant diminution by 30% of the shoot biomass whereas root biomass remained stable. Efficient mycorrhizal symbiosis was maintained during exposure whereas the Rhizobium nodule symbiosis was altered in the surface of soil. By contrast, neither biomass accumulation nor symbiotic association was affected in ryegrass, probably due to a lower sensitivity of this species to PHE exposure. Perspectives of carbon allocation and nitrogen nutrition perturbations are suggested in clovers.     

Predator–prey mass ratio drives microbial activity under dry conditions in Sphagnum peatlands

Mid‐ to high‐latitude peatlands are a major terrestrial carbon stock but become carbon sources during droughts, which are increasingly frequent as a result of climate warming. A critical question within this context is the sensitivity to drought of peatland microbial food webs. Microbiota drive key ecological and biogeochemical processes, but their response to drought is likely to impact these processes. Peatland food webs have, however, been little studied, especially the response of microbial predators. We studied the response of microbial predators (testate amoebae, ciliates, rotifers, and nematodes) living in Sphagnum moss carpet to droughts, and their influence on lower trophic levels and on related microbial enzyme activity. We assessed the impact of reduced water availability on microbial predators in two peatlands using experimental (Linje mire, Poland) and natural (Forbonnet mire, France) water level gradients, reflecting a sudden change in moisture regime (Linje), and a typically drier environment (Forbonnet). The sensitivity of different microbial groups to drought was size dependent; large sized microbiota such as testate amoebae declined most under dry conditions (?41% in Forbonnet and ?80% in Linje). These shifts caused a decrease in the predator–prey mass ratio (PPMR). We related microbial enzymatic activity to PPMR; we found that a decrease in PPMR can have divergent effects on microbial enzymatic activity. In a community adapted to drier conditions, decreasing PPMR stimulated microbial enzyme activity, while in extreme drought experiment, it reduced microbial activity. These results suggest that microbial enzymatic activity resulting from food web structure is optimal only within a certain range of PPMR, and that different trophic mechanisms are involved in the response of peatlands to droughts. Our findings confirm the importance of large microbial consumers living at the surface of peatlands on the functioning of peatlands, and illustrate their value as early warning indicators of change.     

Phenanthrene toxicity and dissipation in rhizosphere of grassland plants (Lolium perenne L. and Trifolium pratense L.) in three spiked soils

Rhizodegradation is a technique involving plants that offers interesting potential to enhance biodegradation of persistent organic pollutants such as polycyclic aromatic hydrocarbons (PAHs). Nevertheless, the behaviour of PAHs in plant rhizosphere, including micro-organisms and the physico-chemical soil properties, still needs to be clarified. The present work proposes to study the toxicity and the dissipation of phenanthrene in three artificially contaminated soils (1 g kg^-1 DW). Experiments were carried out after 2 months of soil aging. They consisted in using different systems with two plant species (Ryegrass—Lolium perenne L. var. Prana and red clover—Trifolium pratense L. var. fourragère Caillard), three kinds of soils (a silty-clay-loam soil “La Bouzule”, a coarse sandy-loam soil “Chenevières” and a fine sandy-loam soil “Maconcourt”). Phenanthrene was quantified by HPLC in the beginning (T ₀) and the end of the experiments (30 days). Plant biomass, microbial communities including mycorrhizal fungi, Rhizobium and PAH degraders were also recorded. Generally phenanthrene contamination did not affect plant biomass. Only the red clover biomass was enhanced in Chenevières and La Bouzule polluted soils. A stimulation of Rhizobium red clover colonisation was quantified in spiked soils whereas a drastic negative phenanthrene effect on the mycorrhization of ryegrass and red clover was recorded. The number of PAH degraders was stimulated by the presence of phenanthrene in all tested soils. Both in ryegrass and red clover planted soils, the highest phenanthrene dissipation due to the rhizosphere was measured in La Bouzule soils. On the contrary, in non-planted soils, La Bouzule soils had also the lowest pollutant dissipation. Thus, in rhizospheric and non-rhizospheric soils the phenanthrene dissipation was found to depend on soil clay content.