Humic-like substances of bacterial origin. I. Some aspects of the formation and nature of humic-like substances produced by Pseudomonas.

Two bacterial strains Pseudomonas acidovorans No 26 and Pseudomonas sp. No 4 grown in Conn and yeast extract-glucose media, or in the media enriched with tyrosine, were found to produce dark brown pigment. It was shown that in the bacterial cultures numerous phenolic and quinone-type compounds were formed and transformed to humic-like polymers. Formation of humic-like substances started in the bacterial cells and was accompanied by the presence of phenyloxidases in the bacterial cultures. The bacterial "humic acids" were obtained from the supernatants in amounts varing from 0.05 to 0.865 mg/1 mg of dry weight of cells and from the cells in amounts of 0.02 to 0.165 mg/1 mg of dry weight of cells, depending on the medium used and time of incubation. The IR spectra of the bacterial "humic acids" appeared to be very similar to IR spectrum of the synthetic humic acids (Fluka A.G.) and contained the same chemical groups as the soil humic acids. The culture medium after growth of the strain No 26 was fractionated into "fulvic, hymatomelanic and humic acid" fractions. The hydrolysates from the obtained fractions contained amino acids and uronic acids. The amino acid composition appeared to be very similar to that of soil humic acids.     

Fluorene and Phenanthrene Uptake and Accumulation by Wheat,Alfalfa and Sunflower from the Contaminated Soil

Polycyclic Aromatic Hydrocarbons (PAHs) are diverse organic contaminants released into the environment by both natural and anthropogenic activities. These compounds have negative impacts on plants growth and development. Although there are many reports on their existence in different parts of plant, their uptake and translocation pathways and mechanisms are not well understood yet. This paper highlights the uptake, translocation and accumulation of PAHs by wheat, sunflower and alfalfa through an experimental study under controlled conditions. Seeds were cultivated in a soil containing 50 mg/kg of phenanthrene and fluorene and their concentrations in plants roots and shoots were determined using a gas chromatograph after 7 and 14 days. The results showed that phenanthrene and fluorene concentrations in the treated plants were increased over the time. PAHs bioavailability was time and species dependent and generally, phenanthrene uptake and translocation was faster than that of fluorene, probably due to their higher K_ow. Fluorene tended to accumulate in roots, but phenanthrene was transported to aerial parts of plants.     

Cs phytoremediation by Sorghum bicolor cultivated in soil and in hydroponic system

Cs accumulation characteristics by Sorghum bicolor were investigated in hydroponic system (Cs level at 50–1000 μmol/L) and in soil (Cs-spiked concentration was 100 and 400 mg/kg soil). Two varieties of S. bicolor Cowly and Nengsi 2# grown on pot soil during the entire growth period (100 days) did not show significant differences on the height, dry weight (DW), and Cs accumulation. S. bicolor showed the potential phytoextraction ability for Cs-contaminated soil with the bioaccumulation factor (BCF) and the translocation factor (TF) values usually higher than 1 in soil system and in hydroponic system. The aerial parts of S. bicolor contributed to 86–92% of the total removed amounts of Cs from soil. Cs level in solution at 100 μmol/L gave the highest BCF and TF values of S. bicolor. Cs at low level tended to transfer to the aerial parts, whereas Cs at high level decreased the transfer ratio from root to shoot. In soil, the plant grew well when Cs spiked level was 100 mg/kg soil, but was inhibited by Cs at 400 mg/kg soil with Cs content in sorghum reaching 1147 mg/kg (roots), 2473 mg/kg (stems), and 2939 mg/kg (leaves). In hydroponic system, average Cs level in sorghum reached 5270 mg/kg (roots) and 4513 mg/kg (aerial parts), without significant damages to its biomass at 30 days after starting Cs treatment. Cs accumulation in sorghum tissues was positively correlated with the metal concentration in medium.     

Soil-derived organic particles and their effects on the community of culturable microorganisms

Soil microbial community interacts with a range of particulate material in the soil, consisting of both inorganic and organic compounds with different levels of water solubility. Though sparingly water-soluble and insoluble organic compounds in the soil may affect living organisms, they are difficult to introduce into microbiological media. Their biological activity (i.e., their effect on soil microorganisms) thus has been almost neglected in most of the cultivation assays. To fill this gap, we propose the use of fine organic particles prepared from soil organic matter that are introduced into a laboratory medium where microbial community is cultivated. To this purpose, submicrometer particles consisting of sparingly water-soluble or insoluble soil organic matter were obtained from humic horizons of two soils by precipitation of organics dissolved in tetrahydrofuran by addition of water. The particles could then be size fractionated by centrifugation, and coarse fraction obtained from humic horizon formed under spruce forest was tested for effects on complex microbial community developing under laboratory conditions. The results indicate that low concentration (20 mg/L) of the particles is efficient to affect the composition of the bacterial community revealed by terminal restriction fragment length polymorphism. The work contributes to understanding the factors that determine the composition of soil microbial community.     

Microbial degradation of polycyclic aromatic hydrocarbons in soils affected by the organic matrix of compost

This paper describes the degradation of naphthalene, phenanthrene, anthracene, fluoranthene, and pyrene in soil and soil/compost mixtures. Compost addition facilitated the degradation of 500 mg naphthalene/kg soil and 100 mg/kg each of other polycyclic aromatic hydrocarbons (PAH) within 25 days in soil systems with water contents below the water-holding capacity. By means of a humic acid extraction, it was demonstrated that the decrease of PAH concentrations after compost addition was not caused by a sorption to organic matter preventing PAH analysis. The enhanced PAH degradation was examined in a series of batch experiments with contaminated soil to evaluate whether the effect of compost addition is caused by the microorganisms of the compost itself, by the properties of the organic matrix of the compost material, or by water-soluble fertilising substances. The experiments revealed that the release of fertilising substances from the compost and the shift of soil pH brought about by the compost did not cause the stimulatory effect. The microorganisms inherent to the compost were also not necessary for the enhanced degradation. Sterilised compost was recolonised by soil microorganisms after a lagphase yielding a degradation activity similar to that of the non-sterilised compost. The presence of the solid organic matrix of the compost seemed to be essential for the enhanced degradation. The soil/compost microflora, which was separated from the organic matrix in liquid cultures, exhibited a much lower degrading activity than in the presence of the solid organic material.     

Effects of the Addition of Humic Substances on Soil Protease Activities in Andosols in the Presence of Cadmium

Cadmium extraction, sorption, and immobilization seem to be the effective mechanisms in detoxification of Cd-contaminated soil. Humic substances present in soils may play an important role both in controlling the negative effects of pollution with Cd and in stabilizing soil enzymes. In this context, we have compared the effects of humic substances on soil protease activities in the presence and absence of Cd in forest and cultivated field soil samples. The samples were taken from surface soils of Andosols in a single upland area of the Kanto district in Japan. Humic substances extracted from the two soils showed differences in elemental composition, the degree of condensation of aromatic groups, and the proportions of major functional groups. Compared with the control soil samples, those with added humic substances showed a significant increase in protease activities, even in the presence of Cd (10 and 50 mg Cd kg^?1 soil). However, the addition of Cd decreased the protease activities, protein contents, and soil pH in both soil samples at each of the two levels of humic substance fortification (+5% and +10%). Moreover, protease activities showed significant negative correlation with exchangeable Cd, but adding humic substances did not lead to a reduction in either sample. Thus, although the addition of humic substances increased and stabilized protease activities, it did not lead to a clear reversal of enzyme inhibition by Cd. The obtained results indicate that in both soil samples the humic substances used in this study did not have sufficient affinity to adsorb Cd, and the impact on enzyme activities depends on the difference in chemical characteristics of the added organic matter, as suggested by the difference in enhancement of protease activities between forest and cultivated field soil samples.     

Pyrene and Phenanthrene Influence on Soil Microbial Populations

Two studies were conducted to evaluate microbial populations in polycyclic aromatic hydrocarbon-contaminated soil. Captina silt loam was freshly exposed to (1) 0 or 2000 mg pyrene/kg and sampled after 10- and 61-wk incubation and (2) 0 or 505 mg pyrene + 445 mg phenanthrene/kg and sampled after a 21-wk incubation. Microbial numbers were determined by plate-count techniques. Isolated bacteria, selected degraders, and wholesoil extracts were analyzed by fatty acid methyl ester analysis (FAME). In the pyrene experiment, pyrene did not affect total bacterial or fungal numbers, but pyrene degraders increased from undetectable levels to 7.09 log₁₀ degraders/g in the contaminated soil. The FAME analysis of bacterial isolates detected no pyrene effect, but wholesoil FAME indicated an increase in the contaminated soil of a fatty acid characteristic of protozoa and a major fatty acid detected in isolated degraders. In the pyrene + phenanthrene experiment, the contaminants had no impact on bacterial, fungal, or actinomycete numbers but increased degrader numbers. No effect of pyrene + phenanthrene was detected by isolate FAME, but whole-soil FAME indicated an effect similar to that in the pyrene experiment. The results indicate that pyrene, although not impacting microbial numbers, may have altered the soil microbial composition and that Captina silt loam can develop an effective degrader population under tested conditions.     

Pyrene and Phenanthrene Influence on Soil Microbial Populations

Two studies were conducted to evaluate microbial populations in polycyclic aromatic hydrocarbon-contaminated soil. Captina silt loam was freshly exposed to (1) 0 or 2000?mg pyrene/kg and sampled after 10- and 61-wk incubation and (2) 0 or 505?mg pyrene + 445?mg phenanthrene/kg and sampled after a 21-wk incubation. Microbial numbers were determined by plate-count techniques. Isolated bacteria, selected degraders, and wholesoil extracts were analyzed by fatty acid methyl ester analysis (FAME). In the pyrene experiment, pyrene did not affect total bacterial or fungal numbers, but pyrene degraders increased from undetectable levels to 7.09 log₁₀ degraders/g in the contaminated soil. The FAME analysis of bacterial isolates detected no pyrene effect, but wholesoil FAME indicated an increase in the contaminated soil of a fatty acid characteristic of protozoa and a major fatty acid detected in isolated degraders. In the pyrene + phenanthrene experiment, the contaminants had no impact on bacterial, fungal, or actinomycete numbers but increased degrader numbers. No effect of pyrene + phenanthrene was detected by isolate FAME, but whole-soil FAME indicated an effect similar to that in the pyrene experiment. The results indicate that pyrene, although not impacting microbial numbers, may have altered the soil microbial composition and that Captina silt loam can develop an effective degrader population under tested conditions.     

Differential decomposition of humic acids by marine and estuarine bacterial communities at varying salinities

Humic substances (HS) constitute 50–80% of total dissolved organic matter (DOM) in freshwaters but in the open ocean <3%, indicating that large fractions of DOM and HS are removed in the estuarine and coastal zone. In order to assess the role of bacteria in this removal, we conducted experiments in flow-through cultures to examine the decomposition of peat bog-born humic acids (HA) by marine (salinity 30, Exp1) and estuarine (salinity 10, Exp2) bacterial communities. After ~70 days 40–>60% of the HA were decomposed. Highest fractions were decomposed in treatments in which the bacterial communities were fed by HA media of a foreign salinity, e.g., of 14 in Exp1 and of 28 in Exp2. Some of the HA material was not decomposed but broken down to smaller moieties entering the fulvic acid (FA) fraction. The analysis of the HA media and their residuals after bacterial decomposition by pyrolysis GC/MS showed that individual organic compounds were decomposed. In Exp1, 32 aliphatic and aromatic compounds, including lignin biomarkers, were detected in the HA medium of which mainly aliphatic compounds were decomposed. In Exp2, 49 compounds were detected of which ~40–60% were not detected any more after bacterial decomposition in the HA fraction but still in the FA fraction. The results show that estuarine and marine bacterial communities can decompose large amounts of HA and that this process is important in reducing the amount of terrestrial HS and DOM entering the estuarine and coastal region.     

蚯蚓对细菌降解土壤中菲的作用

通过30d室内培养试验,分别研究了接种蚯蚓(E)、细菌(B)以及同时接种细菌和蚯蚓(BE)对土壤中菲降解的影响.结果表明: 在土壤中菲的初始污染浓度为50 mg*kg-1的条件下,各处理间菲的降解率差异显著,其降解率的大小顺序依次为:BE》B》E》CK(对照); 在150 mg*kg-1菲的初始污染浓度下,BE处理中菲的降解率高达98.86%,显著高于CK和E处理.B处理中细菌的双加氧酶活性在3种菲初始污染浓度下没有显著差异,而BE处理中双加氧酶的活性随着土壤中菲的初始污染浓度的升高而增加.在相同菲污染浓度下BE处理中蚯蚓体内的菲含量明显高于E处理.表明蚯蚓能够通过生物富集作用降低土壤中菲的浓度,而蚯蚓与细菌的相互作用能够进一步促进土壤中菲的降解.     

Soil Washing Enhanced by Humic Substances and Biodegradable Chelating Agents

Industrial timber treatment sites have resulted in widespread soil contamination by Cu, Cr, and As, presenting potential long-term liability and associated risks to human health and the environment. This study evaluated the roles of natural humic substances (lignite-derived humic substances, standard and commercially available humic acids) and biodegradable chelating agents (ethylenediamine-N,N-disuccinic acid (EDDS) and glutamic-N,N-diacetic acid (GLDA)) for soil washing. Batch kinetic experiments revealed that humic substances promoted Cu extraction at pH 8, but they were significantly adsorbed on the soil at pH 4, possibly posing impediment to soil remediation. The metal extraction by EDDS and GLDA was comparable to that of EDTA (ethylenediamine-tetraacetic acid), and it was more effective at pH 4 than pH 8, probably due to acidic dissolution of metal precipitates and oxides. Metal distribution analysis indicated that the carbonate fraction of Cu and the oxide fraction of As and Cr were mainly extracted, while the exchangeable fraction of Cu increased. The residual leachability tests showed that humic substances reduced the Cu and As leachability but the reduction was insufficient. In contrast, EDDS was able to reduce the leachate concentrations of Cu and As to below 5 mg L^?1, meeting the waste acceptance criteria for landfill disposal. Nevertheless, soil washing methods and remediation strategy may need further modifications to facilitate site restoration and promote soil recycling.     

Residue dynamics and risk assessment of pyraclostrobin in rice,plants, hulls,field soil,and paddy water

A modified quick, easy, cheap, effective, rugged and safe–ultra-performance liquid chromatography–tandem mass spectrometry method was developed and validated for the analysis of pyraclostrobin residue in rice plants, hulls, soil, and paddy water. Pyraclostrobin residue dynamics and final residues were studied in supervised field trials under Good Agricultural Practice conditions in rice, plants, hulls, soil, and paddy water. The levels of detection of all chemicals in five types of matrices were in the ranges of 2 × 10^?7 to 1 × 10^?5 mg/kg with recoveries in the ranges of 74.1–107.4%. The dissipation experiments showed that the half-lives (T_1/2) of pyraclostrobin in the plants, soil, and paddy water were 6.3–13.9, 5.0–34.7, and 6.9–11.6 days, respectively. At pre-harvest intervals (PHIs) of 14, 21, and 28 days, pyraclostrobin residue levels were <0.01–0.19 mg/kg in rice. The residue levels at 35-d PHI were all below 0.1 mg/kg (maximum residue limit of the USA), and all the health risk quotient values calculated were below 0.027. The environment risk of pyraclostrobin in soil and paddy water was also monitored throughout the rice-growing period, and the results showed that the risk level was between low and medium in the soil and high in the paddy water.     

Optimization of Influencing Parameters on Phenanthrene Removal Efficiency in Soil Washing Process by Using Response Surface Methodology

Response surface methodology (RSM) under Box–Behnken design (BBD) was applied to evaluate the effect of the influencing parameters including surfactant concentration, liquid/soil ratio, Humic Acid concentration, and washing time on phenanthrene removal efficiency in soil washing process by using the nonionic surfactant Tween 80 and find an optimal operational conditions to achieve the highest removal efficiency. A polynomial quadratic model was used to correlate phenanthrene removal efficiency and four independent variables (R² = 0.9719). Based on the obtained results the most influential parameter on phenanthrene removal efficiency was surfactant concentration with an impact value of 69.519%. Liquid/soil ratio was also another factor that significantly influenced on removal efficiency with an impact value of 25.014%. The interaction between surfactant concentration and liquid/soil ratio was also shown to have a positive significant effect on removal efficiency (p_value = 0.0027). However, the other independent variables Humic Acid concentration and time were not significant in the ranges selected in this study. Based on the optimization results maximum removal efficiency of 70.692 ± 3.647% was achieved under the conditions of surfactant concentration 5000 mg L^?1, liquid/soil ratio 30 v/w, HA concentration 9.88 mg L^?1, and washing time 2 h, which was in good agreement with predicted value (66.643%).     

The Combined Effect of Cu,Zn and Pb on Enzyme Activities in Soil from the Vicinity of a Wellhead Protection Area

Heavy metals are known to have adverse effects on soil ecosystems, while soil enzyme activities are sensitive to soil pollution. This study investigated the combined effects of Cu, Zn and Pb on the activities of invertase (IN), urease (U) and alkaline phosphatase (ALP) in soil obtained from the vicinity of a wellhead protection area via an orthogonal array (OA) design method. The experimental results showed the following: (1) Cu showed higher inhibition on the activities of all three enzymes than Zn and Pb when three metals were all present in the soil sample. IN activity, U activity and ALP activity decreased as the levels of Cu increased, and ranged from 15.9% to 55.7%, 3.57% to 78.6%, and 3.23% to 75.3%, respectively. Their lowest values were found in samples at 35 days with 400 mg/kg Cu. (2) Zn and Pb had different influences on the activities of the three enzymes. The lowest IN activity (the highest reduction 58.0%) and U activity (76.8%) were observed when Zn was at the concentration of 100 mg/kg after 35 days, whereas the highest inhibitory function of Zn on ALP activity (75.3%) was at 300 mg/kg after 7 days. When the concentration of Pb increased from 35 to 350 mg/kg, the activities of IN (62.5%) and U (69.6%) were most inhibited at 35 days and 14 days, respectively. However, when Pb was at the concentration of 500 mg/kg after 14 days, ALP activity (72.0%) showed the lowest value. (3) With respect to the three hydrolases in this study, ALP was the most sensitive to the two-variable interactive effects of Cu, Zn and Pb, especially Cu?×?Pb. It is concluded that the soil ALP activity may be a sensitive tool for assessing additive toxic effect on soil biochemical parameters. To provide more information about the potential ecological risk of chemicals on soil ecosystems, much more should be done to clearly determine the mechanisms of the combined effects of heavy metals in soil.     

AMF和PGPR联合修复菲和芘污染土壤的效应

丛枝菌根真菌（arbuscular mycorrhizal fungi,AMF）与根围促生细菌（plant growth-promoting rhizobacteria,PGPR）联合降解有毒有机物、修复污染土壤和促进植物生长的作用倍受关注。本试验旨在探究AMF与PGPR联合降解土壤中菲和芘的效应,以菲和芘1:1混合处理浓度各0、50mg/kg、100mg/kg和150mg/kg下对高羊茅Festuca elata接种AMF根内根孢囊霉Rhizophagus intraradices（Ri）、变形球囊霉Glomus versiforme（Gv）、PGPR荧光假单胞菌Pseudomonas fluorescens Ps2-6、芽孢杆菌Bacillus velezensis Ps3-2、Ri+Ps2-6、Ri+Ps3-2、Gv+Ps2-6、Gv+Ps3-2和不接种对照共36个处理。结果表明,供试AMF增加了PGPR的定殖数量;接种PGPR则显著提高AMF的侵染率。AMF、PGPR或AMF+PGPR处理均显著降低土壤中菲和芘含量,促进植物对土壤中菲和芘的吸收,显著提高高羊茅根系和叶片内的菲和芘含量。在土壤中菲和芘100mg/kg和150mg/kg水平下,Gv与Ps2-6及Ri与Ps2-6能相互促进对土壤中菲和芘的去除效应,其中接种Gv+Ps2-6组合处理的去除率最高,达到95%-98%,土壤中多酚氧化酶、脱氢酶和过氧化氢酶活性显著高于单接种处理和不接种对照,而酸性磷酸酶活性变化则表现为相反趋势。其中以Gv+Ps2-6组合处理的多酚氧化酶活性最高,为0.17mg/g,是不接种对照的1.9倍;脱氢酶和过氧化氢酶活性分别达到1.32µg/(g·h)和1.81mL/g;酸性磷酸酶活性则比不接种对照土壤降低27%-45%;易提取球囊霉素相关土壤蛋白含量和总球囊霉素相关土壤蛋白含量分别是不接种对照的1.6倍和1.5倍。     

Effect of Humic Fractions and Clay on Biodegradation of Phenanthrene by a Pseudomonas fluorescens Strain Isolated from Soil

The mineralization of phenanthrene in pure cultures of a Pseudomonas fluorescens strain, isolated from soil, was measured in the presence of soil humic fractions and montmorillonite. Humic acid and clay, either separately or in combination, shortened the acclimation phase. A higher mineralization rate was measured in treatments with humic acid at 100 μg/ml. Humic acid at 10 μg/ml stimulated the transformation only in the presence of 10 g of clay per liter. We suggest that sorption of phenanthrene to these soil components may result in a higher concentration of substrate in the vicinity of the bacterial cells and therefore may increase its bioavailability.     

Degradation of High Molecular Weight PAHs in Contaminated Soil by a Bacterial Consortium: Effects on Microtox and Mutagenicity Bioassays

Bioaugmentation of polycyclic aromatic hydrocarbon (PAH)-contaminated soil was investigated using a mixed bacterial culture (community five) isolated from an abandoned industrial site. Community five was inoculated into contaminated soil containing a total PAH (two- to five-ring compounds) concentration of approximately 820 mg/kg soil. PAH degradation by the indigenous microbial population was restricted to the lower molecular weight compounds (naphthalene, acenaphthene, fluorene and phenanthrene) even with yeast extract addition: these compounds decreased by 14 to 37%, in soil hydrated to 50% water capacity, following 91 days of incubation at 24°C. Inoculation of community five into this PAH-contaminated soil resulted in significant decreases in the concentration of all PAHs over the incubation period: greater than 86% of naphthalene, acenaphthene, fluorene, and phenanthrene were degraded after 91 days, while anthracene, fluoranthene, and pyrene were degraded to lesser extents (51.7 to 57.6%). A lag period of 48 to 63 days was observed before the onset of benz[a]anthracene, benzo[a]pyrene, and dibenz[a,h]anthracene removal. However, significant decreases in the concentration of these compounds (32.6, 25.2, and 18.5%, respectively) were observed after 91 days. No significant decrease in the mutagenic potential of organic soil extracts (as measured by the Ames Test) was observed after incubation of the soil with the indigenous microflora; however, the Microtox toxicity of aqueous soil extracts was reduced sevenfold. In contrast, extracts from contaminated soil inoculated with community five underwent a 43% decrease in mutagenic potential and the toxicity was reduced 170-fold after 91 days incubation. These observations suggest that community five could be utilised for the detoxification of PAH-contaminated soil.     

Column Flushing of Phenanthrene and Copper (II) Co-Contaminants from Sandy Soil Using Tween 80 and Citric Acid

The clean-up of soils co-contaminated with heavy metals and organic compounds is a contemporary issue of remediation efforts. Column flushing was conducted to investigate the performance of nonionic surfactant and/or organic acid solutions, 4000 mg/L Tween 80 (TW80), and/or 0.04 mol/L citric acid (CA), to enhance the simultaneous removal of phenanthrene and copper (II) from the co-contaminated sandy soil. The flushing effects were compared when TW80, CA, TW80 after CA (CA/TW80), CA after TW80 (TW80/CA), and a mixture of TW80 and CA (TW80-CA) were used as flushing agents. The maximum concentrations of phenanthrene in effluent solutions occurred at 3.3, 4.7, 5.3, and 15.3 h during TW80, TW80/CA, TW80-CA, and CA/TW80 flushing and those of copper (II) at 2.7, 3.3, 3.3, and 14.0 h during CA, CA/TW80, TW80-CA, and TW80/CA flushing, respectively. Phenanthrene was mainly desorbed through partitioning into TW80 micelles in aqueous phase while copper (II) was effectively removed through complexation with CA. The removal efficiencies were up to 81.5%, 5.9%, 99.9%, 91.6%, and 99.8% for phenanthrene, and 0.1%, 76.7%, 85.7%, 78.1%, and 84.4% for copper (II) by TW80, CA, TW80/CA, TW80-CA, and CA/TW80. However, it took a long time to use TW80/CA and CA/TW80 to clean phenanthrene and copper (II) efficiently. The overall removal efficiencies of contaminants in the soil column increased with flushing time in the Sigmoidal Model. The results indicated that a combination of TW80 and CA has potential for in situ clean-up of heavy metals and polycyclic aromatic hydrocarbons (PAHs) from co-contaminated soils.     

Selection of cold‐tolerant plants for growth in soils contaminated with organics

A mixture of organic chemicals (MOC) containing equal molar amounts of benzoic acid, hexadecane, 2,2‐dimethyl 4,n‐propyl‐benzene, phenanthrene, pyrene, and either cycloheptane or cis‐decahydronaphthalene (cis‐decalin) was applied to soil at rates of 0 to 8000 mg/kg. In a plant‐screening experiment, growth responses of four legume and five nonlegume species were determined at 10 and 25°C. The MOC applied at 2000 mg/kg reduced the growth of several species without resulting in significant seedling death. At 10°C, the growth of alpine bluegrass (Poa alpina L.) in the 1000 and 2000 mg/kg treatments of soil increased by more than 185%. In a plant growth response experiment, alpine bluegrass and alfalfa (Medicago sativa L.) were grown in soil that had been contaminated at rates of 0 and 2000 mg/kg. At 14 weeks, the shoot and root dry weights of alfalfa were 97% lower in the contaminated soil, while the shoot dry weight, root dry weight, and root length of alpine bluegrass were 135,235 and 268% higher, respectively. Except for pyrene, <23% of the compounds comprising the MOC remained in the soil after 4 weeks and <5% after 14 weeks. The disappearance of the MOC was not significantly influenced by the presence of alfalfa or alpine bluegrass.