Preparation and reactivation of magnetic biochar by molten salt method: Relevant performance for chlorine-containing pesticides abatement

Molten salt has been regarded as a versatile and environmental-friendly method for the material preparation and waste destruction. In this work, molten FeCl3 was utilized for the generation of magnetic biochar (MBC) derived from simultaneous activation and magnetization of biomass. The sample characterization indicated that MBC had a rough surface with BET surface area of 404 m2/g and total pore volume of 0.35cm3/g. Highly dispersed Fe3O4 and nitrogen could be deposited on the surface, leading to an excellent magnetization property. The MBC exhibited a great 2,4-Dichlorophenol (2.4-DCP) and atrazine removal performance in solution with the maximum adsorption capacity achieved 298.12 mg/g and 102.17 mg/g. Kinetics results demonstrated that MBC adsorption met the Pseudo-first-order model better. Molten NaOH-Na2CO3 was provided for the re-activation of exhausted MBC. 2,4-DCP was firstly desorbed from the MBC and subsequently destructed by the active species in the melt medium. Chlorine can be captured in the molten alkaline medium from the XRD pattern of residues.The MBC could be easily recovered with a yield of 98.2% and fixed carbon content of 61.0% after the molten salt regeneration process. With no 2,4-DCP detected, 65.5% and 31.69% of initial Cl was found in washing water and residues with the molten NaOH-Na2CO3, respectively. After 4 cycles of regeneration and adsorption, 60.55%-72.22% of initial adsorption capacity can be kept. This preparation and regeneration method can be an effective way to reduce the risk of secondary pollution of chlorinated organic compounds during adsorbent regeneration.

Implications: Molten salt (MS) is a salt or multiple salts with a low melting point, and has been applied in many sectors and is regarded as a crucial role in terms of energy, environmental, and resource sustainability. In our paper, magnetic biochar was prepared by one-step activation and magnetization of fir dust using molten FeCl₃?6H₂O. Meanwhile, a regeneration method using molten alkaline salt was provided. Magnetic biochar generated in our study performed well in the 2,4-dichlorophenol and atrazine adsorption. After four cycles of regeneration and adsorption, 72.2% of initial 2,4-DCP adsorption capacity can be kept.     

Comparison of microbial indicators under two water regimes in a soil amended with combined paper mill sludge and decomposed cow manure

An incubation study was conducted under laboratory conditions to compare the effects of soil amendment of combined paper mill sludge (PS) and decomposed cow manure (DCM) on selected microbial indicators. A lateritic soil (Typic Haplustalf) was amended with 0 (control), 20 or 80tha(-1) (wet weight) of PS or DCM. The amended soils were then adjusted to 60% water holding capacity (WHC) or submerged conditions, and incubated at 27 degrees C in dark for up to 120days (d). The microbial biomass C (MBC), the basal soil respiration and the enzyme activities of the beta-glucosidase, acid phosphatase and sulphatase were analyzed at day 15, 30, 45, 60 and 120. Compared to the unamended soil (control), the MBC, the basal soil respiration and the enzyme activities increased with the rate of PS and DCM. At similar rate, the DCM treatment increased significantly the MBC, the soil respiration and the enzyme activities compared to the PS treatment. Also, the water regimes affected the microbial activities. At 60% WHC, the MBC and soil respiration increased during the first 30d and decreased thereafter. The enzyme activities showed similar trends, where they increased for the first 60d, and decreased thereafter. In contrast, under submerged condition, the MBC and enzymes activities declined during 120d, whereas the soil respiration increased. Compared to the control, the used of PS and DCM had no negative impact of the soil microbial parameters, even at the highest application rate. Long-term field experiments are required to confirm these laboratory results.     

Microbial biomass and enzyme activities in submerged rice soil amended with municipal solid waste compost and decomposed cow manure

We studied the suitability of municipal solid waste compost (MSWC) application to submerged rice paddies in the perspective of metal pollution hazards associated with such materials. Experiments were conducted during the wet seasons of 1997, 1998 and 1999 on rice grown under submerged condition, at the Agriculture Experimental Farm, Calcutta University at Baruipur, West Bengal, India. The treatments consisted of control, no input; MSWC, at 60 kgNha(-1); well decomposed cow manure (DCM), at 60 kgNha(-1); MSWC (30 kgNha(-1)) +Urea (30 kgNha(-1)); DCM (30 kgNha(-1)) +U (30 kgNha(-1)) and Fertilizer, (at 60:30:30 NPK kgha(-1) through urea, single superphosphate and muriate of potash respectively). Soil microbial biomass-C (MBC), MBC as percentage of organic-C (ratio index value, RIV), urease and acid phosphatase activities were higher in DCM than MSWC-treated soils, due to higher amount of biogenic organic materials like water soluble organic carbon, carbohydrate and mineralizable nitrogen in the former. The studied parameters were higher when urea was integrated with DCM or MSWC, compared to their single applications. Soil MBC, urease and acid phosphatase activities periodically declined up to 60 day after transplanting (DAT) and then increased after crop harvest. The heavy metals in MSWC did not detrimentally influence MBC, urease and acid phosphatase activities of soil. In the event of long term MSWC application, changes in soil quality parameters should be monitored regularly, since heavy metals once entering into soil persist over a long period.     

Assessment of the impact of pesticide residues on microbiological and biochemical parameters of tea garden soils in India

The main aim of this study was to assess the impact of pesticidal residues on soil microbial and biochemical parameters of the tea garden soils. The microbial biomass carbon (MBC), basal (BSR) and substrate induced respirations (SIR), β -glucosidase activity and fluorescein diacetate hydrolyzing activity (FDHA) of six tea garden soils, along with two adjacent forest soils (control) in West Bengal, India were measured. The biomass and its activities and biochemical parameters were generally lower in the tea garden soils than the control soils. The MBC of the soils ranged from 295.5 to 767.5 μ g g^{? 1}. The BSR and SIR ranged from 1.65 to 3.08 μ g CO₂-C g^{? 1} soil h^{? 1} and 3.08 to 10.76 μ g CO₂-C g^{? 1}h^{? 1} respectively. The β -glucosidase and FDHA of the soils varied from 33.3 and 76.3 μ g para-nitrophenol g^{? 1} soil h^{? 1} and 60.5 to 173.5 μ g fluorescein g^{? 1}h^{? 1}respectively. The tea garden soils contained variable residues of organophosphorus and organochlorine pesticides, which negatively affected the MBC, BSR, SIR, FDHA and β -glucosidase activity. Ethion and chlorpyriphos pesticide residues in all the tea garden soils varied from 5.00 to 527.8 ppb and 17.6 to 478.1 ppb respectively. The α endosulfan, β endosulfan and endosulfan sulfate pesticide residues in the tea garden soils ranged from 7.40 to 81.40 ppb, 8.50 to 256.1 ppb and 55 to 95.9 ppb respectively. Canonical correlation analysis shows that 93% of the total variation was associated with the negative impact of chlorpyriphos, β and α endosulfan and endosulfan sulfate on MBC, BSR and FDHA. At the same time ethion had negative impact on SIR and β -glucosidase. Data demonstrated that the pesticide residues had a strong impact on the microbial and biochemical components of soil quality.     

Assessment of the impact of pesticide residues on microbiological and biochemical parameters of tea garden soils in India

The main aim of this study was to assess the impact of pesticidal residues on soil microbial and biochemical parameters of the tea garden soils. The microbial biomass carbon (MBC), basal (BSR) and substrate induced respirations (SIR), beta-glucosidase activity and fluorescein diacetate hydrolyzing activity (FDHA) of six tea garden soils, along with two adjacent forest soils (control) in West Bengal, India were measured. The biomass and its activities and biochemical parameters were generally lower in the tea garden soils than the control soils. The MBC of the soils ranged from 295.5 to 767.5 micro g g(- 1). The BSR and SIR ranged from 1.65 to 3.08 mu g CO2-C g(- 1) soil h(- 1) and 3.08 to 10.76 micro g CO2-C g(- 1)h(- 1) respectively. The beta-glucosidase and FDHA of the soils varied from 33.3 and 76.3 micro g para-nitrophenol g(- 1) soil h(- 1) and 60.5 to 173.5 micro g fluorescein g(- 1)h(- 1)respectively. The tea garden soils contained variable residues of organophosphorus and organochlorine pesticides, which negatively affected the MBC, BSR, SIR, FDHA and beta -glucosidase activity. Ethion and chlorpyriphos pesticide residues in all the tea garden soils varied from 5.00 to 527.8 ppb and 17.6 to 478.1 ppb respectively. The alpha endosulfan, beta endosulfan and endosulfan sulfate pesticide residues in the tea garden soils ranged from 7.40 to 81.40 ppb, 8.50 to 256.1 ppb and 55 to 95.9 ppb respectively. Canonical correlation analysis shows that 93% of the total variation was associated with the negative impact of chlorpyriphos, beta and alpha endosulfan and endosulfan sulfate on MBC, BSR and FDHA. At the same time ethion had negative impact on SIR and beta-glucosidase. Data demonstrated that the pesticide residues had a strong impact on the microbial and biochemical components of soil quality.     

Soil carbon and nitrogen dynamics in the first year following herbicide and scalping in a revegetation trial in south-east Queensland,Australia

During revegetation, the maintenance of soil carbon (C) pools and nitrogen (N) availability is considered essential for soil fertility and this study aimed to evaluate contrasting methods of site preparation (herbicide and scalping) with respect to the effects on soil organic matter (SOM) during the critical early establishment phase. Soil total C (TC), total N (TN), hot-water extractable organic C (HWEOC), hot-water extractable total N (HWETN), microbial biomass C and N (MBC and MBN), total inorganic N (TIN) and potentially mineralizable N (PMN) were measured over 53 weeks. MBC and MBN were the only variables affected by herbicide application. Scalping caused an immediate reduction in all variables, and the values remained low without any sign of recovery for the period of the study. The impact of scalping on HWETN and TIN lasted 22 weeks and stabilised afterwards. MBC and MBN were affected by both herbicide and scalping after initial treatment application and remained lower than control during the period of the study but did not decrease over time. While scalping had an inevitable impact on all soil properties that were measured, that impact did not worsen over time, and actually improved plant growth (unpublished data) while reducing site establishment costs. Therefore, it provides a useful alternative for weed control in revegetation projects where it is applied only once at site establishment and where SOM would be expected to recover as canopy closure is obtained and nutrient cycling through litterfall commences.     

Effects of Cd and Pb on soil microbial community structure and activities

Background, aim, and scope  

Soil contamination with heavy metals occurs as a result of both anthropogenic and natural activities. Heavy metals could have long-term hazardous impacts on the health of soil ecosystems and adverse influences on soil biological processes. Soil enzymatic activities are recognized as sensors towards any natural and anthropogenic disturbance occurring in the soil ecosystem. Similarly, microbial biomass carbon (MBC) is also considered as one of the important soil biological activities frequently influenced by heavy metal contamination. The polymerase chain reaction–denaturing gradient gel electrophoresis (DGGE) has recently been used to investigate changes in soil microbial community composition in response to environmental stresses. Soil microbial community structure and activities are difficult to elucidate using single monitoring approach; therefore, for a better insight and complete depiction of the soil microbial situation, different approaches need to be used. This study was conducted in a greenhouse for a period of 12 weeks to evaluate the changes in indigenous microbial community structure and activities in the soil amended with different application rates of Cd, Pb, and Cd/Pb mix. In a field environment, soil is contaminated with single or mixed heavy metals; so that, in this research, we used the selected metals in both single and mixed forms at different application rates and investigated their toxic effects on microbial community structure and activities, using soil enzyme assays, plate counting, and advanced molecular DGGE technique. Soil microbial activities, including acid phosphatase (ACP), urease (URE), and MBC, and microbial community structure were studied.     

Fractionation and bioavailability of metals and their impacts on microbial properties in sewage irrigated soil

A study was conducted to evaluate the effect of long-term irrigation of sewage contaminated with heavy metals like Cd, Cr, Cu and Pb on microbial and biochemical parameters of soils of West Bengal, India. The microbial parameters included microbial biomass carbon (MBC), microbial metabolic quotient; the biochemical parameters included fluorescein diacetate hydrolyzing activity, beta-glucosidase, urease, phosphatase, and aryl sulphatase activities. A sequential extraction technique was used to quantify water soluble, exchangeable, carbonate bound, Fe/Mn-oxide bound, organically bound, and residual metal fractions. Metal concentrations in the two most labile fractions (i.e., water soluble and exchangeable fractions) were generally low. Total metal concentrations at each site seemed to be associated with soil amorphous Fe and Al minerals. The MBC and the enzymes studied were significantly and negatively correlated with water soluble and exchangeable metals but not significantly correlated with other forms, indicating that water soluble and exchangeable forms exerted a strong inhibitory effect on the soil microbial and biochemical parameters. It was concluded that irrigating soils with metal contaminated sewage seemed to damage soil quality in the long term.     

Comparative bioremediation potential of four rhizospheric microbial species against lindane

Four microbial species (Kocuria rhizophila, Microbacterium resistens, Staphylococcus equorum and Staphylococcus cohnii subspecies urealyticus) were isolated from the rhizospheric zone of selected plants growing in a lindane contaminated environment and acclimatized in lindane spiked media (5-100 μg mL⁻¹). The isolated species were inoculated with soil containing 5, 50 and 100 mg kg⁻¹ of lindane and incubated at room temperature. Soil samples were collected periodically to evaluate the microbial dissipation kinetics, dissipation rate, residual lindane concentration and microbial biomass carbon (MBC). There was a marked difference (p < 0.05) in the MBC content and lindane dissipation rate of microbial isolates cultured in three different lindane concentrations. Further, the dissipation rate tended to decrease with increasing lindane concentrations. After 45 d, the residual lindane concentrations in three different spiked soils were reduced to 0%, 41% and 33%, respectively. Among the four species, S. cohnii subspecies urealyticus exhibited maximum dissipation (41.65 mg kg⁻¹) and can be exploited for the in situ remediation of low to medium level lindane contaminated soils.     

Response of microbial activities in two contrasting soils to 4-nonylphenol treated with biosolids

In the application of biosolids on agricultural lands, 4-nonylphenol (4-NP) in soils is an important environmental concern because of its associated estrogenic risk to animals and human beings. Incubation experiments that involved the mixing of two contrasting soils (A: calcareous sandy soil; B: acidic clayey soil) and biosolids in 4-NP were performed to examine the effect of 4-NP on the rate of production of CO2, the mineralization of N and the microbial biomass, by considering the biodegradation of 4-NP for the evaluation of soil health. The experimental results indicated that the half-life (t1/2) of 4-NP increased with the supplementary concentration of 4-NP (80, 160 and 240 mg kg(-1)) in the two soils, and the t1/2 values in the soil A are always lower than that in soil B. The 4-NP supplement in the biosolids reverses C mineralization in soil B more than it does in soil A, but it reverses N mineralization in soil A more than in soil B. The aeration status and microbial population of the biosolids treated soils are key factors in determining the time course of 4-NP degradation associated with the microbial activities. The 4-NP was biodegraded mainly by bacteria, and the effect on C and N mineralization of 4-NP input is determined by a balance of the reductions in microbial biomass C (MBC) and N (MBN). After destruction in microbial cell membrane and protein structures by the 4-NP, C and N mineralization, MBC and MBN were subsequently followed by a final decline phase for the later period of incubation.     

Effect of Pencycuron on Microbial Parameters of Waterlogged Soil

A laboratory study was conducted to monitor the effect of pencycuron [1-(4-chlorobenzyl)-1-cyclopentyl-3-phenylurea] on microbial parameters of alluvial (AL) soil (Typic udifluvent) and coastal saline (CS) soil (Typic endoaquept) under waterlogged condition. Pencycuron at field rate (FR), 2FR and 10FR affected the microbial biomass C (MBC), soil ergosterol content and fluorescein diacetate hydrolyzing activity (FDHA) differentially. The DCM amendment did not seem to have any counteractive effect on the toxicity of pencycuron on the microbial variables. The change in microbial metabolic quotient (qCO₂) and microbial respiration quotient (Q_R), indicated pencycuron induced disturbance at 10FR. Present study revealed that the metabolically activated microbial population was more suppressed compared to the dormant population.     

The effect of initial concentration, co-application and repeated applications on pesticide degradation in a biobed mixture

A 180 d laboratory experiment was conducted to investigate the degradation rates of chlorpyrifos (10 and 50 mg kg(-1)) and metalaxyl (100 mg kg(-1)) separately and co-applied in a biomix constituted by topsoil, vine-branches and urban-waste-garden compost. The effect of repeated application of metalaxyl was also investigated. Microbial biomass-C (MBC) content and metabolic quotient (qCO(2)) were measured to evaluate changes in microbial biomass size and activity induced by the presence of the two pesticides. Degradation rate decreased with increasing concentration of chlorpyrifos in all treatments. Metalaxyl half-life was significantly reduced in co-application with chlorpyrifos indicating a synergic interaction between the two pesticides in favour of enhanced degradation rate for metalaxyl but not for chlorpyrifos. Furthermore, repeated application resulted in a sharp reduction of metalaxyl half-life from 37 d after first application to 4 d after third application. MBC content was negatively influenced by the addition of pesticides but it started to recover immediately, in both separate and co-applied treatments, reaching the control value when pesticide residues were about 50% of the initial concentration. The qCO(2) reached a steady-state after about 20 d in separately applied and 40 d in co-applied treatments, indicating a tendency to arrive at a new metabolic equilibrium. In conclusion, the biomix tested has been shown to degrade pesticides relatively fast and to have a microbial community that is varied enough to allow selection of those microorganisms able to degrade metalaxyl and chlorpyrifos.     

Carbon mineralization,microbial activity and metal dynamics in tailing ponds amended with pig slurry and marble waste

A field experiment was set up in Cartagena-La Unión Mining District, SE Spain, aimed at evaluating the short-term effects of pig slurry (PS) amendment alone and together with marble waste (MW) on organic matter mineralization, microbial activity and stabilization of heavy metals in two tailing ponds. These structures pose environmental risk owing to high metals contents, low organic matter and nutrients, and null vegetation. Carbon mineralization, exchangeable metals and microbiological properties were monitored during 67 d. The application of amendments led to a rapid decrease of exchangeable metals concentrations, except for Cu, with decreases up to 98%, 75% and 97% for Cd, Pb and Zn, respectively. The combined addition of MW + PS was the treatment with greater reduction in metals concentrations. The addition of PS caused a significant increase in respiration rates, although in MW + PS plots respiration was lower than in PS plots. The mineralized C from the pig slurry was low, approximately 25–30% and 4–12% for PS and MW + PS treatments, respectively. Soluble carbon (Csol), microbial biomass carbon (MBC) and β-galactosidase and β-glucosidase activities increased after the application of the organic amendment. However, after 3 d these parameters started a decreasing trend reaching similar values than control from approximately day 25 for Csol and MBC. The PS treatment promoted highest values in enzyme activities, which remained high upon time. Arylesterase activity increased in the MW + PS treatment. Thus, the remediation techniques used improved soil microbiological status and reduced metal availability. The combined application of PS + MW reduced the degradability of the organic compounds.     

Effect of pencycuron on microbial parameters of waterlogged soil

A laboratory study was conducted to monitor the effect of pencycuron [1-(4-chlorobenzyl)-1-cyclopentyl-3-phenylurea] on microbial parameters of alluvial (AL) soil (Typic udifluvent) and coastal saline (CS) soil (Typic endoaquept) under waterlogged condition. Pencycuron at field rate (FR), 2FR and 10FR affected the microbial biomass C (MBC), soil ergosterol content and fluorescein diacetate hydrolyzing activity (FDHA) differentially. The DCM amendment did not seem to have any counteractive effect on the toxicity of pencycuron on the microbial variables. The change in microbial metabolic quotient (qCO2) and microbial respiration quotient (QR), indicated pencycuron induced disturbance at 10FR. Present study revealed that the metabolically activated microbial population was more suppressed compared to the dormant population.     

不同利用方式土壤中磷的吸附与解吸特性

研究不同利用方式土壤对磷的吸附解吸特性及其影响因素。采用室内恒温培养法研究了湖南典型土壤(红壤、潮土和紫色土)的不同利用方式(旱地、水田)土壤对P的吸附和解吸过程。土壤对磷的吸附解吸过程是分阶段进行的,用Langmuir方程拟合程度比Freundlich方程高,相关系数均在0.90(P<0.05)以上;从最大吸附量(Qm)、吸附反应常数(K)和最大缓冲容量(MBC)3项吸附参数综合考虑,旱地对P的吸附无论在强度还是容量方面均大于同母质的水田土壤;解吸率随着吸附量的增加而增大,其大小为旱地<水田。Qm与<0.002 mm粘粒及无定形铁含量呈正相关,相关系数分别为0.95和0.94(P<0.05)。不同利用方式土壤P吸附解吸特性差别较大,针对不同土壤应采取不同的磷素管理措施以实现作物增产和保护环境的双重效益。     

Thermal co-reduction approach to vary size of silver nanoparticle: its microbial and cellular toxicology

In recent years, silver nanoparticles (AgNPs) have attracted considerable interest in the field of food, agriculture and pharmaceuticals mainly due to its antibacterial activity. AgNPs have also been reported to possess toxic behavior. The toxicological behavior of nanomaterials largely depends on its size and shape which ultimately depend on synthetic protocol. A systematic and detailed analysis for size variation of AgNP by thermal co-reduction approach and its efficacy toward microbial and cellular toxicological behavior is presented here. With the focus to explore the size-dependent toxicological variation, two different-sized NPs have been synthesized, i.e., 60 nm (Ag60) and 85 nm (Ag85). A detailed microbial toxicological evaluation has been performed by analyzing minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), diameter of inhibition zone (DIZ), growth kinetics (GrK), and death kinetics (DeK). Comparative cytotoxicological behavior was analyzed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. It has been concluded by this study that the size of AgNPs can be varied, by varying the concentration of reactants and temperature called as “thermal co-reduction” approach, which is one of the suitable approaches to meet the same. Also, the smaller AgNP has shown more microbial and cellular toxicity.     

Photochemical behaviour of carbendazim in aqueous solution

To elucidate the photochemical behaviour of carbendazim (or MBC) in superficial waters, photolysis studies have been carried out in aqueous solutions at several pH using a UV light source (high pressure mercury arc lamp) or a solar light simulator (xenon arc lamp). The kinetics of photodecomposition of carbendazim was determined using HPLC-DAD and the identification of photoproducts was carried out with HPLC-MS (ESI negative and positive mode). According to the experimental results carbendazim is a rather stable molecule in the dark or in environmental conditions. The pH influence of the environmental medium on the photodegradation rate has been confirmed. The photochemical process can be considerably accelerated in alkaline solutions using HPK-quartz irradiation (quantum efficiency at pH 9 phi = 3.1 x 10(-3) degraded molecule per absorbed photon) while the photodegradation is not as efficient under a simulated sun irradiation (quantum efficiency in the suntest phi = 10(-4) at pH 7). Three photoproducts have been tentatively identified in pure water: 2-aminobenzimidazole, benzimidazole isocyanate and monocarbomethoxy-guanidine (issued from the cleavage of the benzimidazole ring). The last one seems very stable and could be accumulated in the environment.     

Pencycuron application to soils: degradation and effect on microbiological parameters

Clay loam soil from agricultural fields of alluvial (AL) soil (typic udifluvent) and coastal saline (CS) soil (typic endoaquept) were investigated for the degradation and effect of pencycuron application at field rate (FR), 2-times FR (2FR) and 10-times FR (10FR) with and without decomposed cow manure (DCM) on soil microbial variables under laboratory conditions. Pencycuron degraded faster in CS soil and in soil amended with DCM. Pencycuron spiking at FR and 2FR resulted in a short-lived (in case of 10FR slightly longer) and transitory toxic effect on soil microbial biomass-C (MBC), ergosterol content and fluorescein diacetate hydrolyzing activity (FDHA). Amendment of DCM did not seem to have any counteractive effect of the toxicity of pencycuron on the microbial variables. The ecophysiological status of the soil microbial communities as expressed by microbial metabolic quotient (qCO2) and microbial respiration quotient (Q(R)) changed, but for a short period, indicating pencycuron induced disturbance. The duration of this disturbance was slightly longer at 10FR. Pencycuron was more toxic to the metabolically activated soil microbial populations, specifically the fungi. It is concluded that side effects of pencycuron at 10FR on the microbial variables studied were only short-lived and probably of little ecological significance.     

In vitro toxicity of fungicides with different modes of action to alfalfa anthracnose fungus,Colletotrichum destructivum

Abstract

Sensitivity of 24 isolates of Colletotrichum destructivum O’Gara, collected from alfalfa plants in Serbia, to eight selected fungicides, was investigated in this study. Molecular identification and pathogenicity test of isolates tested were also performed. Fungicide sensitivity was evaluated in vitro, using mycelial growth assay method. All isolates exhibited significant pathogenicity, causing necrosis at the alfalfa seedling root tips two days after inoculation. Using the primer pair GSF1-SR1 and by comparing the amplified fragments of the tested isolates with the marker (M), the presence of the amplicon of the expected size of about 900?bp was determined for all isolates. The isolates tested in this study showed different sensitivity towards fungicides in vitro. Mycelial growth was highly inhibited by QoI (quinone outside inhibitors) fungicide pyraclostrobin (mean EC₅₀=0.39?µg mL^?1) and by DMI (demethylation-inhibiting) fungicide tebuconazole (mean EC₅₀=0.61?µg mL^?1), followed by azoxystrobin (mean EC₅₀=2.83?µg mL^?1) and flutriafol (mean EC₅₀=2.11?µg mL^?1). Multi-site fungicide chlorothalonil and MBC (methyl benzimidazole carbamate) fungicide thiophanate-methyl evinced moderate inhibition with mean EC₅₀=35.31 and 62.83?µg mL^?1, respectively. Thirteen isolates were sensitive to SDHI (succinate dehydrogenase inhibitors) fungicide boscalid and fluxapyroxad, (mean EC₅₀=0.49 and 0.19?µg mL^?1, respectively), while the rest of isolates were highly resistant.     

Influence of root-exudates concentration on pyrene degradation and soil microbial characteristics in pyrene contaminated soil

The effect of ryegrass (Lolium perenne L.) root-exudates concentration on pyrene degradation and the microbial ecological characteristics in the pyrene contaminated soil was investigated by simulating a gradually reducing concentration of root exudates with the distance away from root surface in the rhizosphere. Results showed that, after the root-exudates were added 15 d, the pyrene residue in contaminated soil responded nonlinearly in the soils with the same pyrene contaminated level as the added root-exudates concentration increased, which decreased first and increased latter with the increase of the added root-exudates concentration. The lowest pyrene concentration appeared when the root exudates concentration of 32.75 mg kg(-1) total organic carbon (TOC) was added. At the same time, changes of microbial biomass carbon (MBC, C(mic)) and microbial quotient (C(mic)/C(org)) were opposite to the trend of pyrene degradation as the added root-exudates concentration increased. Phospholipid fatty acid (PLFA) analysis revealed that bacteria was the dominating microbial community in pyrene contaminated soil, and the changing trends of pyrene degradation and bacteria number were the same. The changing trend of endoenzyme-dehydrogenase activity was in accordance with that of soil microbe, indicating which could reflect the quantitative characteristic of detoxification to pyrene by soil microbe. The changes in the soils microbial community and corresponding microbial biochemistry characteristics were the ecological mechanism influencing pyrene degradation with increasing concentration of the added root-exudates in the pyrene contaminated soil.