Interactions between the salt marsh grass Spartina patens, arbuscular mycorrhizal fungi and sediment bacteria during the growing season

The interaction between the salt marsh plant Spartina patens, arbuscular mycorrhizal fungi (AMF) and bacteria in salt marsh sediment was examined in a long-term arbuscular mycorrhizas (AM) suppression study by applying the systemic fungicide benomyl to field-collected sediment cores with and without S. patens plants. Microbial populations were sampled four times corresponding to major plant phenological stages (dormancy, vegetative growth, reproduction, and senescence) previously linked to changes in microbial populations under field conditions. Benomyl-treatment of soil cores significantly suppressed AM colonization on S. patens, keeping values relatively consistent throughout the growing season (11.5%) whereas plants in non-treated cores experienced seasonal increases and declines in AM colonization (26.6% during vegetative growth to 11.5% during dormancy). Soil physicochemical parameters were not affected by benomyl application. In unvegetated cores, no benomyl- or seasonal effects were displayed by cell numbers and specific biomass of DAPI-stained organisms, members of the domain bacteria and here especially members of the α-, β-, γ- and δ-subdivisions of proteobacteria that were the most abundant bacterial groups. In vegetated cores, the microbial community as well as specific bacterial populations were at least twice as large in terms of number and biomass than in samples from unvegetated cores with significant seasonal changes for DAPI-stained cells, for members of the domain bacteria and for members of the α- and γ-subdivisions of proteobacteria. In benomyl-treated cores, the population of γ-subdivision of proteobacteria was significantly smaller than in non-treated cores, and a positive association was found between this bacterial group and root length colonized by AM suggesting that AM-suppression can affect populations of specific soil bacterial populations in salt marsh sediment. Benomyl-treatment had no effect on the diversity of N-fixing bacteria as evidenced by PCR-RFLP analysis, but seasonal changes were noted in vegetated cores with populations during active plant growth substantially different from populations during dormancy and senescence.     

Spatio-temporal variability of microbial abundance and community structure in the puddled layer of a paddy soil cultivated with wetland rice (Oryza sativa L.)

The puddled layer of paddy soils represents a highly dynamic environment regarding the spatio-temporal variability of biogeochemical conditions. To study these effects on the abundance and community structure of microbial populations, a rhizotron experiment was conducted throughout an entire growing season of wetland rice. Soil samples were taken from selected areas of the puddled layer (bulk soil, oxidized layer, rhizosphere) at main plant developmental stages such as (i) the initial stage, (ii) tillering, (iii) panicle initiation, (iv) flowering, and (v) maturity. Cell numbers of archaea, bacteria, and selected phyla were assessed by catalyzed reporter deposition-fluorescence in situ hybridization (CARD-FISH). The structure and diversity of microbial communities was analyzed by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) along with sequencing of selected bands. Following submergence of the paddy soil, shifts of bacterial community structure were observed in the oxidized layer and the rhizosphere. Members of the β-Proteobacteria became predominant in the rhizosphere at tillering stage and were affiliated with aerobic, iron-oxidizing bacteria of the genus Sideroxydans. Seasonal effects were mainly visible in the rhizosphere, as several phylogenetic subgroups including methanotrophic bacteria showed increased cell numbers at flowering stage. Cell numbers of methanogenic archaea were also highest at flowering stage (bulk soil, rhizosphere) and members of the Methanocellales were identified as predominant archaeal populations in areas of oxic and anoxic conditions. In contrast to bacteria, the communities of archaea in the puddled layer of the studied paddy soil were less influenced by spatio-temporal variations of biogeochemical conditions.     

Degradation of an Atrazine and Metolachlor Herbicide Mixture in Pesticide-Contaminated Soils from Two Agrochemical Dealerships in Iowa

The fate of atrazine and metolachlor,applied as a mixture, in soil taken from twopesticide-contaminated sites in Iowa (denoted as Alphaor Bravo) were determined in laboratory studies. Atrazine and metolachlor degradation, as well asatrazine mineralization, were greater in soilcollected from Kochia scoparia L. (Schrader)rhizosphere than in soils from unvegetated areas. Theradiolabeled ¹⁴C-carbinol and¹⁴C-morpholinone metabolites were identified in¹⁴C-metolachlor-applied soil 60 d aftertreatment. The half-life for atrazine in Alpha soilwas significantly less in the rhizosphere soil (50 d)than in unvegetated soil (193 d). Quantities ofspecific atrazine degraders were one to two orders ofmagnitude greater in Bravo soils than in Alpha soils. In an experiment with plants present, significantlymore ¹⁴C-atrazine was taken up by K.scoparia (9.9% of the applied ¹⁴C) than by Brassica napus L. Significantly less atrazine wasextractable from soils vegetated with K.scoparia than from soils vegetated with B.napus or unvegetated soils.     

Influence of Three Aquatic Macrophytes on Mitigation of Nitrogen Species from Agricultural Runoff

Agricultural runoff containing nitrogen fertilizer is a major contributor to eutrophication in aquatic systems. One method of decreasing amounts of nitrogen entering rivers or lakes is the transport of runoff through vegetated drainage ditches. Vegetated drainage ditches can enhance the mitigation of nutrients from runoff; however, the efficiency of nitrogen removal can vary between plant species. The efficiency of three aquatic macrophytes, cutgrass (Leersia oryzoides), cattail (Typha latifolia), and bur-reed (Sparganium americanum), to mitigate dissolved and total nitrogen from water was investigated. Replicate mesocosms of each plant species were exposed to flowing water enriched with ammonium and nitrate for 6?h, allowed to remain stagnant for 42?h, and then flushed with non-enriched water for an additional 6?h to simulate a second storm event. After termination of the final simulated runoff, all vegetated treatments lowered total nitrogen loads exiting mesocosms by greater than 50%, significantly more than unvegetated controls, which only decreased concentrations by 26.9% (p????0.0023). L. oryzoides and T. latifolia were more efficient at lowering dissolved nitrogen, decreasing ammonium by 42?±?9% and 59?±?4% and nitrate by 67?±?6% and 64?±?7%, respectively. All treatments decreased ammonium and nitrate concentrations within mesocosms by more than 86% after 1?week. However, T. latifolia and L. oryzoides absorbed nitrogen more rapidly, lowering concentrations by greater than 98% within 48?h. By determining the nitrogen mitigation efficiency of different vegetative species, plant communities in agricultural drainage ditches can be managed to significantly increase their remediation potential.     

The Effects of Cattails (Typha latifolia L.) on Concentrations and Partitioning of Metals in Surficial Sediments of Surface-Flow Constructed Wetlands

The concentration and partitioning of metals in vegetatedand unvegetated surface sediments were compared among fourwetlands designed for wastewater treatment. In wetlands witha significant depth gradient, the highest concentrations ofmetals were found in deeper unvegetated sediments. When theeffect of depth was removed, surface sediments with andwithout cattails (Typha latifolia L.) had similarconcentrations of total metals (Fe, Mn, Zn and Cu). However,vegetated sediments of all four wetlands had higher organiccontent (p < 0.01). Furthermore, the redox potential wassignificantly lower (p < 0.01) in vegetated sediments for 3out of 4 wetlands. Vegetated and unvegetated sediments werealso compared based on geochemical fractions (reactive,silicate, persistent organic and pyritic). Generally, halfof the total metal concentration in sediments was associatedwith the reactive fraction (metal oxides, monosulfides andadsorbed onto organic matter). Zn was an exception as halfof the total Zn was associated with the persistent organicfraction. The partitioning of metals in shallow vegetatedsediments differed from deep unvegetated sediments at theMonahan and Falconbridge wetlands. In contrast, there was nosignificant difference in the partitioning of metals insurface sediments with or without cattails at the Panel andRiverwalk wetlands. This study suggests that the effect ofcattails on the bulk concentration and partitioning ofmetals may be minimal in surface-flow constructed wetlands.     

Diversity, abundance and vertical distribution of methane-oxidizing bacteria (methanotrophs) in the sediments of the Xianghai wetland, Songnen Plain, northeast China

Purpose

Methanotrophs in wetlands are of great importance because up to 90 % of the methane (CH₄) produced in such wetlands could be oxidized by methanotrophs before reaching the atmosphere. The Xianghai wetland of Songnen Plain represents an important ecosystem in northeast China. However, methanotrophic characteristics in this ecosystem have not been studied in detail. The aim of this study is to give an overview of methanotrophic diversity and vertical distribution in the sediments of this important wetland.

Materials and methods

Sediment cores were collected from three freshwater marshes, each dominated by a particular vegetation type: Carex alata, Phragmites australis and Typha orientalis. The diversity of methanotrophs was studied by phylogenetic analysis of both the 16S rRNA gene and the particulate methane monooxygenase (pmoA) gene. Methanotroph abundance was determined by quantitative PCR (qPCR) targeting the pmoA gene; group-specific pmoA gene quantification was also used to estimate the abundance of each methanotrophic group.

Results and discussion

16S rRNA and pmoA gene homological analysis revealed the presence of type Ia, Ib and II methanotrophs. Novel pmoA sequences distantly affiliated to cultured Methylococcus sp. were detected, implying the existence of novel methanotrophs in the wetland. Most obtained representatives of Methylobacter genus (both 16S rRNA and pmoA genes) were closely clustered in relation to sequences acquired from the Zoige wetland, Tibet and Siberia permafrost soils, therefore suggesting methanotrophs belonging to Methylobacter genus shared characteristics with methanotrophs in cold areas. The dominance of type I methanotrophs (especially the Methylobacter genus) was detected by both clone library analysis and group-specific qPCR assay. The relatively high methanotroph diversity and pmoA copy numbers measured in the T. orientalis marsh sediments indicated that vegetation type played an important role during CH₄ oxidation in the wetland.

Conclusions

We present the first data set on methanotroph diversity and vertical distribution in the sediments of the Xianghai wetland. DNA sequences information of Methylococcus-like methanotrophs in the wetland will facilitate the isolating of novel methanotrophs from the wetland. In a worldwide context, our study has enriched the database of genotypic diversity of methanotrophs, which will help in the understanding of the geographical distribution of methanotrophic communities.     

Laboratory mesocosm studies of Fe,Al, Mn,Ca, and Mg dynamics in wetlands exposed to synthetic acid coal mine drainage

To evaluate the potential for constructed wetlands to treat acid coal mine drainage, six model wetland mesocosms (each 2.4 m × 15 cm) were filled with Sphagnum peat (15 cm deep), planted either with cattails (Typha latifolia) and living Sphagnum, living Sphagnum only, or left as bare peat (2 mesocosms per treatment). The model wetlands were exposed to synthetic acid coal mine drainage (pH 3.5, concentrations of Fe²⁺, Al³⁺, Mn^2*, Ca²⁺, and Mg²⁺ of 78.8, 10.0, 5.2, 12.0, and 4.5 mg L^?1, respectively) at a rate of 90 mL min^?1, 6 hr d^?1, 5 d wk^?1, over a 16 week period. Chemical analysis of peat at periodic intervals indicated that the model wetlands were net sources of Al³⁺, Mn²⁺ Ca²⁺ and Mg²⁺, but net sinks for Fe²⁺. Type of vegetation had no significant effect on Fe²⁺ retention; of the 204 g of Fe²⁺ added to the model wetland systems, 162 g were retained. Formation of Fe oxides accounted for 73 to 86% of the Fee' retention, with exchangeable Fe contributing 0.2 to 1.2%, organically bound Fe contributing 4 to 19%, and residual Fe contributing 7 to 15% of total Fee' retention. Fe retention was greatest at the inflow ends of the model wetlands where Fe retention appeared to reach saturation at a final Fe concentration in the peat of 235 mg g^?1. At the rate of application of the synthetic acid mine drainage, we estimated that the model wetland systems would have reached complete Fe saturation after 157 days. We suggest that the mesocosm approach could be useful in generating site-specific data that can be applied to the formulation of cost-benefit analyses that can compare a proposed wetland treatment system with alternative conventional chemical methods for treating acid mine drainage.     

Bacterial phylogenetic diversity in a constructed wetland system treating acid coal mine drainage

Microorganisms in acid mine drainage are typically acidophiles that mediate the oxidation of reduced compounds of iron and sulfur. However, microbial populations in wetland systems constructed to treat acid mine drainage are not well characterized. The purpose of this study was to analyze bacterial diversity, using cultivation-independent molecular ecological techniques, in a constructed wetland that received acid drainage from an abandoned underground coal mine. DNA was purified from Fe(III)-precipitates from the oxidized surface zone of wetland sediments and 16S rRNA gene sequences were amplified and cloned. A total of 200 clones were analyzed by restriction fragment length polymorphism (RFLP) and 77 unique RFLP patterns were obtained with four restriction enzymes. Of these patterns, 30 most dominant unique clones were selected for sequencing of their 16S rRNA genes. Half of these 30 clones could be matched with autotrophic iron- and sulfur-oxidizing bacteria (Acidithiobacillus ferrooxidans and Acidithiobacillus thiooxidans). Several clones also formed a clade with heterotrophic iron-oxidizing bacteria (TRA2-10, TRA3-20, and TRA5-3) and heterotrophic bacteria (Stenotrophomas maltophilia, Bordetella spp., Alcaligenes sp., Alcaligenes faecalis, and Alcaligenes xylosoxidans). Approximately 40% and 35% of the analyzed RFLP restriction patterns were consistent with A. ferrooxidans and A. thiooxidans, respectively. The relatively high frequency of acidithiobacilli is consistent with the chemical and physical characteristics of this site—i.e., continuous, abundant supply of reduced iron and sulfur compounds, pH 3–4, ambient temperature, and limited organics originating from the coal seam and from vegetation or soil surrounding the inlet channel to the wetland. The RFLP results were consistent with our previous culture-independent PCR-DGGE and FISH study, showing relatively low bacterial diversity and predominance of mesophilic acidithiobacilli in oxic wetland sediments.     

Nesting habitat selection by bitterns Botaurus stellaris in Britain and the implications for wetland management

The rarity and decline of the bittern Botaurus stellaris in Britain has prompted large-scale wetland restoration and more recently, wetland creation projects. In order to guide such habitat management, we investigated whether any large or fine-scale features within British wetland sites best described the selection of female bittern nesting positions. Birds nested in continuous vegetation (usually Phragmites dominated) that was on average 100 m at its narrowest width. When compared with random locations, nests had less scrub and more vegetated open water edge in their wider vicinity and were immediately surrounded by smaller percentage coverage of non-Phragmites species and thicker Phragmites stems. Of most importance, female bitterns nested at points where deeper water was maintained into the driest part of the season, perhaps using the presence of water tolerant plant species as an indication of this. The results can be used to aid the design of new wetland sites to take into account the needs of nesting female bittern.     

Methane concentrations and methanotrophic community structure influence the response of soil methane oxidation to nitrogen content in a temperate forest

Methane oxidation in forest soils removes atmospheric CH₄. Many studies have determined methane uptake rates and their controlling variables, yet the microorganisms involved have rarely been assessed simultaneously over the longer term. We measured methane uptake rates and the community structure of methanotrophic bacteria in temperate forest soil (sandy clay loam) on a monthly basis for two years in South Korea. Methane uptake rates at the field site did not show any seasonal patterns, and net uptake occurred throughout both years. In situ uptake rates and uptake potentials determined in the laboratory were 2.92 ± 4.07 mg CH₄ m⁻² day⁻¹ and 51.6 ± 45.8 ng CH₄ g⁻¹ soil day⁻¹, respectively. Contrary to results from other studies, in situ oxidation rates were positively correlated with soil nitrate concentrations. Short-term experimental nitrate addition (0.20-1.95 μg N g⁻¹ soil) significantly stimulated oxidation rates under low methane concentrations (1.7-2.0 ppmv CH₄), but significantly inhibited oxidation under high methane concentrations (300 ppmv CH₄). We analyzed the community structures of methanotrophic bacteria using a DNA-based fingerprinting method (T-RFLP). Type II methanotrophs dominated under low methane concentrations while Type I methanotrophs dominated under high methane concentrations. Nitrogen addition selectively inhibited Type I methanotrophic bacteria. Overall, the results of this study indicate that the effects of inorganic N on methane uptake depend on methane concentrations and that such a response is related to the dissimilar activation or inhibition of different types of methanotrophic bacteria.     

Nutrient Cycling and Retention Along a Littoral Gradient in a Dutch Shallow Lake in Relation to Water Level Regime

Littoral zones are characterized by gradients in depth and vegetation biomass, influencing nutrient retention capacity. A field experiment was conducted in a Phragmites australis dominated littoral zone to investigate nutrient retention and its effect on surface water quality. Measurements were done in mesocosms where water levels could be manipulated. Nutrient status was investigated along a gradient perpendicular to the shore during two growing seasons, one with a stable water level and one with a gradually decreasing water level. Nutrient concentrations in sediment, soil pore water and surface water were significantly lower in the vegetated than in the unvegetated zone. The negative correlations of nutrients in sediment and water, with nutrient contents of the vegetation suggest a direct effect of the vegetation. Nutrient uptake and biomass of the vegetation was higher in continuously flooded soils than in seasonally emerging sediments higher along the littoral gradient, probably due to the increased salinity in drained zones. Denitrification rate was highest in the unvegetated zone and was positively related to water level. Flooded littoral zones did result in a higher nutrient retention than drained zones. On small scale, for an optimal nutrient retention a fluctuating regime is not necessarily better suited than a stable water level, but on a larger scale it can substantially increase the width of the vegetated zone. It is important to optimize conditions for helophyte growth since the positive effect of vegetation on nutrient retention, at least at local scale, has been demonstrated in this study.     

Impact of 12-year field treatments with organic and inorganic fertilizers on crop productivity and mycorrhizal community structure

The effect of manure and mineral fertilization on the arbuscular mycorrhizal (AM) fungal community structure of sunflower (Helianthus annuus L.) plants was studied. Soils were collected from a field experiment treated for 12 years with equivalent nitrogen (N) doses of inorganic N, dairy manure slurry, or without N fertilization. Fresh roots of tall fescue (Festuca arundinacea Schreb.) grass collected from the field plots without N fertilization and unfumigated field soils were used as native microbial inoculum sources. Sunflower plants were sown in pots containing these soils, and three different means of manipulating the microbial community were set: unfumigated soil with fresh grass roots, fumigated soil with fresh grass roots, or fumigated soil with sterilized grass roots. Assessing the implications with respect to plant productivity and mycorrhizal community structure was investigated. Twelve AM fungal OTUs were identified from root or soil samples as different taxa of Acaulospora, Claroideoglomus, Funneliformis, Rhizophagus, and uncultured Glomus, using PCR-DGGE and sequencing of an 18S rRNA gene fragment. Sunflower plants grown in manure-fertilized soils had a distinct AMF community structure from plants either fertilized with mineral N or unfertilized, with an abundance of Rhizophagus intraradices-like (B2). The results also showed that AM inoculation increased P and N contents in inorganic N-fertilized or unfertilized plants, but not in manure-fertilized plants.     

Assessing effects of earthworm cast on methanotrophic community in a soil biocover by concurrent use of microarray and quantitative real-time PCR

Effects of earthworm cast (EC) on the methanotrophic community in a soil biocover were evaluated using microarray and quantitative real-time PCR (qRT-PCR). Soil was collected from a biocover with either soil alone or a mixture of soil and EC (3:1, w/w). The microarray results showed a more diverse methanotrophic community in the EC biocover than that in the soil biocover (p < 0.05). A principal component analysis result confirmed a substantial change in the methanotrophic community structure due to the added EC. Type I methanotrophs dominated both biocovers, with Methylobacter being most abundant. The qRT-PCR results showed that EC greatly increased the methanotrophic population levels (p < 0.05) up to 100 fold. In conclusion, EC can increase the population density of methanotrophs as well as their diversity, resulting in a substantial shift in the community structure. The results confirmed the promising potential of EC as a bed material for enhancing the biocover performance.     

The differential effects of ammonium and nitrate on methanotrophs in rice field soil

It has been known that nitrogenous fertilizers can either stimulate or inhibit methane oxidation in soils. The mechanism, however, remains unclear. Here we conducted laboratory incubation experiments to evaluate the effects of ammonium versus nitrate amendment on CH₄ oxidation in a rice field soil. The results showed that both N forms stimulated CH₄ oxidation. But nitrate stimulated CH₄ oxidation to a greater extent than ammonium per unit N base. The 16S rRNA genes and the pmoA genes were analyzed to determine the dynamics of total bacterial and methanotrophic populations, respectively. The methanotrophic community consisted of type I and type II methanotrophs and was dominated by type I group after two weeks of incubation. Nitrate promoted both types of methanotrophs, but ammonium promoted only type I. DNA-based stable isotope probing confirmed that ammonium stimulated the incorporation of ¹³CH₄ into type I methanotrophs but not type II, while nitrate caused almost homogenous distribution of ¹³CH₄ in type I and type II methanotrophs. Our study suggests that nitrate can promote CH₄ oxidation more significantly than ammonium and is probably a better N source for both types of methanotrophs in rice field soil. More investigations, e.g. using ¹⁵N labeling, are necessary to elucidate this possibility.     

Soil methane oxidation and methanotroph responses to afforestation of pastures with Pinus radiata stands

Afforestation of pastures in New Zealand reduces methane (CH₄) production from soil, while also stimulating oxidation of atmospheric CH₄ by soil methanotrophs. However, neither the mechanisms by which soil CH₄ oxidation is enhanced by afforestation, nor how long after forest planting tree-dependent responses in CH₄ oxidation become detectable are fully known. Here, we investigated the effects of different-aged stands (5-20 y) of the exotic pine (Pinus radiata (D. Don)) on CH₄ oxidation and methanotrophic community structure in soils, compared with adjacent, long-established pastures. Two of the pastures were on volcanic soils and two were on non-volcanic soils. Although the CH₄ fluxes in soils from these young stands were not significantly different from those in the associated pastures, the rate of oxidation of added ¹³CH₄ was higher in the pine soils. Both fluxes and ¹³CH₄ oxidation rates were higher in the volcanic than the non-volcanic soils. Combined phospholipid fatty acid (PLFA) and stable isotope probe (SIP) analyses suggested that type II methanotrophs (PLFA C18:1ω7) were most active in all soils followed by uncultivable bacteria (C17:0ai). Molecular analysis of the methanotrophic community structure using pmoA (particulate methane monooxygenase) genes suggested that a particular type II methanotroph (TRF 35) was dominant in all soils, but more so in the pine than in pasture soils. A type I methanotroph (TRF 245) was more prevalent in the pasture than in associated pine soils, whereas TRF 128 (a type II methanotroph) was slightly more dominant in soils under pine. Cloning and sequencing data suggest TRFs 35 and 128, which differ from one another, belong to distant relatives of Methylocapsa sp; TRF 245 is related to Methylococcus capsulatus. Land-use change resulted in changes in soil bulk density, porosity, moisture contents and in methanotrophic community structure. Methane oxidation rates were most closely related to soil moisture, as well as to the methanotrophic community structure, and nitrate-N, extractable C and total C concentrations. Stepwise multiple regression also suggested a weak effect (P = 0.06) of stand age on CH₄ oxidation rate. By contrast, the responses of the methanotrophic community structure to this land-use change were more readily detected by the specific molecular analyses, and indicated a predominance of type II methanotrophs in pine soils.     

Fluid Catalytic Cracking Unit Emissions and Their Impact

A constructed wetland composed of a pond- and a marsh-type wetland was employed to remove nitrogen (N) and phosphorus (P) from effluent of a secondary wastewater treatment plant in Korea. Nutrient concentrations in inflow water and outflow water were monitored around 50 times over a 1-year period. To simulate N and P dynamics in a pond- and a marsh-type wetland, mesocosm experiments were conducted. In the field monitoring, ammonium (NH ₄ ⁺ ) decreased from 4.6 to 1.7 mg L^?1, nitrate (NO ₃ ^? ) decreased from 6.8 to 5.3 mg L^?1, total N (TN) decreased from 14.6 to 10.1 mg L^?1, and total P (TP) decreased from 1.6 to 1.1 mg L^?1. Average removal efficiencies (loading basis) for NO ₃ ^? , NH ₄ ⁺ , TN, and TP were over 70%. Of the environmental variables we considered, water temperature exhibited significant positive correlations with removal rates for the nutrients except for NH ₄ ⁺ . Results from mesocosm experiments indicated that NH ₄ ⁺ was removed similarly in both pond- and marsh-type mesocosms within 1 day, but that NO ₃ ^? was removed more efficiently in marsh-type mesocosms, which required a longer retention time (2?C4 days). Phosphorus was significantly removed similarly in both pond- and marsh-type mesocosms within 1 day. Based on the results, we infer that wetland system composed of a pond- and a marsh-type wetland consecutively can enhance nutrient removal efficiency compared with mono-type wetland. The reason is that removal of NH ₄ ⁺ and P can be maximized in the pond while NO ₃ ^? requiring longer retention time can be removed through both pond and marsh. Overall results of this study suggest that a constructed wetland composed of a pond- and a marsh-type wetland is highly effective for the removal of N and P from effluents of a secondary wastewater treatment plant.     

Characterization and taxonomical note about Thai Erianthus germplasm collection: the morphology, flowering phenology and biogeography among E. procerus and three types of E. arundinaceus

Erianthus, one of the genus in Saccharum complex, is important genetic resources for sugarcane improvement. The morphology and flowering phenology of 108 accessions belonging to Erianthus procerus and three types of E. arundinaceus collected from throughout Thailand were compared. PCA analysis based on 22 characteristics clearly supported the separation of Type II and Type III of E. arundinaceus from E. procerus and Type I of E. arundinaceus according to their morphological characteristics, particularly their bud size, and the development of root primordia, which greatly affected axis I of the PCA analysis. E. procerus and Type I showed overlapping in many of their characteristics including their flowering periods. Flower characteristics such as rachis joint length, which were used for previous taxonomic classifications, were not available for the classification of the Thai samples because of the wide variation and overlapping among them. Most of these phenotype similarities and differences are considered to have developed convergently as a result of niche adaptation. Type II and III inhabit riverbanks and streambeds where floods occur frequently, while E. procerus and Type I mainly grow in non-flooding areas such as mountainous grassland, the edge of forests, and beside fields. All Thai Erianthus show unique geographic distributions in Thailand. In particular, the biogeographic boundary between Type II and Type III appeared to be located at the Isthmus of Kra. Although some types showed morphological similarities, reproductive isolation among the four groups seemed to be maintained by differentiation in geographic distribution, habitat preference, and flowering timing.     

Potential for Phosphate Mitigation from Agricultural Runoff by Three Aquatic Macrophytes

Phosphate from agricultural runoff is a major contributor to eutrophication in aquatic systems. Vegetated drainage ditches lining agricultural fields have been investigated for their potential to mitigate runoff, acting similarly to a wetland as they filter contaminants. It is hypothesized that some aquatic macrophytes will be more effective at removing phosphate than others. In a mesocosm study, three aquatic macrophyte species, cutgrass (Leersia oryzoides), cattail (Typha latifolia), and bur-reed (Sparganium americanum), were investigated for their ability to mitigate phosphate from water. Mesocosms were exposed to flowing phosphate-enriched water (10?mg?L^?1) for 6?h, left stagnant for 42?h, and then flushed with non-nutrient enriched water for an additional 6?h to simulate flushing effects of a second storm event. Both L. oryzoides and T. latifolia decreased the load of dissolved phosphate (DP) in outflows by greater than 50?%, significantly more than S. americanum, which only decreased DP by 15?±?6?% (p????0.002). All treatments decreased concentrations inside mesocosms by 90?% or more after 1?week, though the decrease occurred more rapidly in T. latifolia and L. oryzoides mesocosms. By discovering which species are better at mitigating phosphate in agricultural runoff, planning the community composition of vegetation in drainage ditches and constructed wetlands can be improved for optimal remediation results.     

Crude Oil-Contaminated Soil Phytoremediation by Using Cyperus brevifolius (Rottb.) Hassk

The degradation of total oil and grease (TOG) in crude oil-contaminated soil in the presence of Cyperus brevifolius (Rottb.) Hassk was investigated in a net house study. C. brevifolius plants were transplanted in to spiked soil containing 8% (w/w) crude oil. The capability of plant for enhancing the biodegradation process was tested in pots containing fertilized and unfertilized soil over a 360-day period. Analysis of the degradation of hydrocarbon contaminants, plant growth, and biomass was conducted at 60-day interval. In the presence of contaminants, plant biomass and height were significantly reduced. The specific root surface area was reduced under the effects of crude oil. Concerning TOG content in soil, C. brevifolius could decrease up to 86.2% in TA (crude oil-contaminated soil with fertilizer) and 61.2% in TC (crude oil-contaminated soil without fertilizer). In the unvegetated pots, the reduction of TOG was 13.7% in TB (crude oil-contaminated soil with fertilizer) and 12.5% in TD (crude oil-contaminated soil without fertilizer). However, biodegradation was significantly more in vegetated pots than in unvegetated pots (p?=?0.05). The addition of fertilizer had positive effect on TOG degradation in the presence of C. brevifolius compared to the unfertilized treatments. Thus, there was evidence of C. brevifolius enhancing the biodegradation of crude oil in soil under the conditions of this experiment.     

Diversity and antagonistic potential of Pseudomonas spp. associated to the rhizosphere of maize grown in a subtropical organic farm

Pseudomonas spp. are one of the most important bacteria inhabiting the rhizosphere of diverse crop plants and have been frequently reported as biological control agents (BCAs). In this work, the diversity and antagonistic potential of Pseudomonas spp. in the rhizosphere of maize cultivars Nitroflint and Nitrodent grown at an organic farm in Brazil was studied by means of culture-dependent and -independent methods, respectively. Sampling of rhizosphere soil took place at three different stages of plant development: 20, 40 and 106 days after sowing. A PCR-DGGE strategy was used to generate specific Pseudomonas spp. fingerprints of 16S rRNA genes amplified from total community rhizosphere DNA. Shifts in the relative abundance of dominant populations (i.e. PCR-DGGE ribotypes) along plant development were detected. A few PCR-DGGE ribotypes were shown to display cultivar-dependent relative abundance. No significant differences in diversity measures of DGGE fingerprints were observed for different maize cultivars and sampling times. The characterisation and assessment of the antagonistic potential of a group of 142 fluorescent Pseudomonas isolated from the rhizosphere of both maize cultivars were carried out. Isolates were phenotypically and genotypically characterised and screened for in vitro antagonism towards three phytopathogenic fungi and the phytopathogenic bacterium Ralstonia solanacearum. Anti-fungal activity was displayed by 13 fluorescent isolates while 40 isolates were antagonistic towards R. solanacearum. High genotypic and phenotypic diversity was estimated for antagonistic fluorescent Pseudomonas spp. PCR-DGGE ribotypes displayed by antagonists matched dominant ribotypes of Pseudomonas DGGE fingerprints, suggesting that antagonists may belong to major Pseudomonas populations in the maize rhizosphere. Antagonists differing in their genotypic and phenotypic characteristics shared the same DGGE electrophoretic mobility, indicating that an enormous genotypic and functional diversity might be hidden behind one single DGGE band. Cloning and sequencing was performed for a DGGE double-band which had no corresponding PCR-DGGE ribotypes among the antagonists. Sequences derived from this band were affiliated to Pseudomonas stutzeri and P. alcaligenes 16S rRNA gene sequences. As used in this study, the combination of culture-dependent and -independent methods has proven to be a powerful tool to relate functional and structural diversity of Pseudomonas spp. in the rhizosphere.