Study Potential of Indigenous <Emphasis Type="Italic">Pseudomonas aeruginosa</Emphasis> and <Emphasis Type="Italic">Bacillus subtilis</Emphasis> in Bioremediation of Diesel-Contaminated Water

Petroleum products which are used in a wide variety of industries as energy sources and raw materials have become a major concern in pollution of terrestrial and marine environments. The purpose of this study was to assess the potential of indigenous microbial isolates for degradation of diesel fuel. Two most proficient bacterial strains among five isolated strains from polluted soil of an industrial refinery were studied. The isolates then were identified as Pseudomonas aeruginosa and Bacillus subtilis using biochemical tests and 16S rRNA gene sequence analyses. P. aeruginosa showed higher biodegradation efficiency than B. subtilis in shaking flask containing diesel-contaminated water. P. aeruginosa and B. subtilis degraded about 87 and 75% of total hydrocarbons, respectively, in flasks containing 2% diesel and 98% water. The biodegradation efficiency of the isolates decreased as diesel contamination increased from 2 to 5%. The isolates showed significantly higher efficiency on degradation of short-chain hydrocarbons in 20 days, i.e., by using P. aeruginosa, removal efficiency of C₁₀ hydrocarbons was near 90%, while about 69% of C₂₀₊ hydrocarbons and 47% of aromatic hydrocarbons were removed. Therefore, the isolates showed high capability in biodegradation of diesel contamination of the refinery.     

Tolerance and Bioaccumulation of Arsenate by <Emphasis Type="Italic">Aspergillus Oryzae</Emphasis> TLWK-09 Isolated from Arsenic-Contaminated Soils

Eleven indigenous arsenic-tolerant fungi were isolated from arsenic-contaminated mine tailing and identified by molecular biology methods. Among them, Aspergillus oryzae (denoted as A. oryzae TLWK-09) had high tolerance and bioaccumulation of As(V). The maximum tolerance to As(V) concentration of A. oryzae TLWK-09 reached 5000 mg/L. As(V) bioaccumulation on A. oryzae TLWK-09 in the aqueous system was investigated under different environmental conditions such as mycelia dosage, contact time, pH, and ionic strength. Bioaccumulation data of As(V) were fitted to Langmuir model, and the maximum uptake capacity of A. oryzae TLWK-09 for As(V) was 54.12 mg/g at 301 K. The morphological structures of mycelia changed obviously under As(V) stress by scanning electron microscope (SEM) and transmission electron microscope (TEM) analysis. The analysis of Fourier transform infrared spectroscopy (FTIR) indicated the presence of carboxyl, hydroxyl, and amino groups on the fungal mycelia, which showed that these groups accounted for As(V) bioaccumulation. These results suggested that A. oryzae TLWK-09 could be an efficient and promising bioremediation material for As(V) pollution.     

Manganese (II) Removal from Aqueous Solutions by <Emphasis Type="Italic">Bacillus cereus</Emphasis>: an Optimization Study Using Experimental Design and Response Surface Methodology

The present study was carried out in order to investigate the removal of soluble Mn²⁺ from an aqueous solution using Bacillus cereus. A manganese aqueous solution at 50 mg L^?1 was treated, and the product was less than 1 mg L^?1 of residual concentration, which complied with environmental regulations. Before the optimization, B. cereus was able to remove Mn²⁺ ions from an aqueous solution; however, the residual content was around 2.5 mg L^?1. Screening experiments aiming at defining the effects of the growth medium composition indicated that both casamino acid-peptone and yeast extract contributed to manganese removal. These experiments also showed the interaction between these two components of the culture media, nevertheless the use of glucose did not prove significant. Considering these observations, the Doehlert design was used to generate a response surface. The model was significant with the p value lower than 0.05 and the lack-of-fit not significant (p > 0.05). The optimized composition of the growth medium was defined as 0.5 g L^?1 of casamino acid-peptone and 0.25 g L^?1 of yeast extract, whereas glucose could be removed from the original growth medium. When the optimized condition of the growth medium was used, the time required for manganese removal was reduced from 21 to 8 days. After optimization, B. cereus was capable of producing high manganese removal, reducing the residual concentration to levels below 1.0 mg L^?1.     

Combined bioremediation of soil co-contaminated with cadmium and endosulfan by <Emphasis Type="Italic">Pleurotus eryngii</Emphasis> and <Emphasis Type="Italic">Coprinus comatus</Emphasis>

Purpose

The subjects of this study were to investigate the remediating potential of the co-cultivation of Pleurotus eryngii and Coprinus comatus on soil that is co-contaminated with heavy metal (cadmium (Cd)) and organic pollutant (endosulfan), and the effects of the co-cultivated mushrooms on soil biochemical indicators, such as laccase enzyme activity and bacterial counts.

Materials and methods

A pot experiment was conducted to investigate the combined bioremediation effects on co-contaminated soil. After the mature fruiting bodies were harvested from each pot, the biomass of mushrooms was recorded. In addition, bacterial counts and laccase enzyme activity in soil were determined. The content of Cd in mushrooms and soil was detected by the flame atomic absorption spectrometry (FAAS), and the variations of Cd fractions in soil were determined following the modified BCR sequential extraction procedure. Besides, the residual endosulfan in soil was detected by gas chromatography-mass spectrometry (GC-MS).

Results and discussion

The results indicated that co-cultivation of P. eryngii and C. comatus exerted the best remediation effect on the co-contaminated soil. The biomass of mushroom in the co-cultivated group (T group) was 1.57–13.20 and 19.75–56.64% higher than the group individually cultivated with P. eryngii (P group) or C. comatus (C group), respectively. The concentrations of Cd in the fruiting bodies of mushrooms were 1.83–3.06, 1.04–2.28, and 0.67–2.60 mg/kg in T, P, and C groups, respectively. Besides, the removal rates of endosulfan in all treatments exceeded 87%. The best bioremediation effect in T group might be caused by the mutual promotion of these two kinds of mushrooms.

Conclusions

The biomass of mushroom, laccase activity, bacterial counts, and Cd content in mushrooms were significantly enhanced, and the dissipation effect of endosulfan was slightly higher in the co-cultivated group than in the individually cultivated groups. In this study, the effect of co-cultivated macro fungi P. eryngii and C. comatus on the remediation of Cd and endosulfan co-contaminated soil was firstly reported, and the results are important for a better understanding of the co-remediation for co-contaminated soil.

     

Biodegradation Mechanism of Phenanthrene by Halophilic <Emphasis Type="Italic">Hortaea</Emphasis> sp. B15

This aim of the study is to investigate a halophilic bacterium Hortaea sp. B15, isolated from petroleum-contaminated soil for biodegradation of phenanthrene. Hortaea sp. B15 has the ability to completely degrade phenanthrene (100 mg/L) under salinity 10% within 1-week incubation. The metabolitic product of phenanthrene was identified and assayed by using ultraviolet-visible spectrophotometer and mass spectral analysis. Result revealed that Hortaea sp. B15 metabolized phenanthrene to form 9,10-phenanthrene quinone, salicylic acid, and gentisic acid. Hortaea sp. B15 has an efficient utilization of phenanthrene in high-saline liquid medium. All the results indicated that the fungus has a promising application for the study of high-molecular-weight PAH biodegradation and contaminated saline-alkali soil bioremediation.     

The impact of adsorption on the exposure assessment of <Emphasis Type="Italic">Bacillus thuringiensis</Emphasis> in the water-sediment system: a mathematical model approach

A model to assess the environmental fate of bacteria to be used as plant protection products or biocides is presented. The model, based on a black-box approach, takes into account the bacterial cell adsorption on the sediment phase and the degradation kinetics and allows the calculation of the expected environmental density of bacterial cells and the predicted environmental concentration of bacterial toxins in the water-sediment system. An example of calculation results is reported for the study cases of Bacillus thuringiensis subsp. aizawai strain GC-91, B. thuringiensis subsp. kurstaki strain ABTS-351, and B. thuringiensis subsp. tenebrionis strain NB 176, used as plant protection products, and B. thuringiensis subsp. israelensis strains AM 65–52 and SA3A, used as biocides. The calculation of expected environmental density and predicted environmental concentration for bacteria in soil-water systems allows to achieve an appropriate risk assessment of the environmental fate and of the impact of bacterial pest control agents or bacterial biocides on non-target organisms.     

Assessing the potential of using biochar in mine rehabilitation under elevated atmospheric CO<Subscript>2</Subscript> concentration

Purpose

Re-establishment of soil nitrogen (N) capital is a priority in mine rehabilitation. We aimed to evaluate the effects of biochar addition on improving mine spoil N pools and the influence of elevated CO₂ concentration on mine rehabilitation.

Materials and methods

We assessed the effects of pinewood biochar, produced at three temperatures (650, 750 and 850 °C, referred as B₆₅₀, B₇₅₀ and B₈₅₀, respectively), on mine spoil total N concentrations with five different plant species, including a tree species (Eucalyptus crebra), N-fixing shrubs (Acacia floribunda and Allocasuarina littoralis) and C₃ and C₄ grasses (Austrodanthonia tenuior and Themeda australis) incubated at ambient (400 μL L^?1) and elevated (700 μL L^?1) atmospheric CO₂ concentrations, as well as the effects of elevated CO₂ on mine rehabilitation.

Results and discussion

Soil total N significantly improved following biochar incorporation under all plant species (P < 0.05) except for T. Australis. E. crebra had the highest soil total N (0.197%, 0.198% and 0.212% for B₆₅₀, B₇₅₀ and B₈₅₀, respectively). Different from the negligible influence of elevated CO₂ on soil properties under the grasses and the N-fixing shrubs, elevated CO₂ significantly increased soil water and hot water extractable organic C (WEOC and HWEOC, respectively) and decreased total C under E. crebra, indicating that the nutrient demands were not met.

Conclusions

Biochar addition showed the potential in mine rehabilitation in terms of improving soil N pool, especially with E. crebra. However, it would be more difficulty to rehabilitate mine spoils in future with the rising atmospheric CO₂ concentration.

     

An efficient approach towards the bioremediation of copper,cobalt and nickel contaminated field samples

Purpose

The synthetic soil based bioremediation approach as reasonable and sustainable practice at the farming level where desired bioremediation could be established at lower cost.

Materials and methods

Metal-tolerant bacteria from different environmental field samples, (a) a municipal dump site, (b) an agricultural field and (c) sludge of electro-plating industries, were screened and characterized. Bioremediation of metal contaminants through isolated bacteria was compared under two different conditions, synthetic soil and basic minimal media containing copper, cobalt and nickel.

Results and discussion

The pollutants arising from industrial effluents are imparting a huge negative impact on agricultural land. Microbes are predominant in heavy metal-contaminated sites, which signifies as a potential opportunity for the researchers towards bioremediation. Three bacterial species showed high metal tolerance; 16S ribosomal DNA (rDNA) analysis revealed that the organisms were Proteus vulgaris strain, Stenotrophomonas sp. and Bacillus thuringiensis. Percentage removal of metals was also analysed under different concentrations and pH.

Conclusions

The current tested methods are helpful in streamlining the natural compliance of fragile elements and its uptake into the microbial system under in vitro and in situ conditions.

     

Hydrodynamic Conditions Influence Bacterial Growth and Phenol Biodegradation of Strains with Different Morphology and Motility

Microorganisms are frequently exposed to flowing fluid, thus to investigate bacterial characteristics under different hydrodynamic conditions is of great importance in microbial ecology. This study characterized bacterial growth and phenol biodegradation of three strains, i.e., Microbacterium oxydans (rod-shaped, non-motile), Alcaligenes faecalis (rod-shaped, motile), and Staphylococcus haemolyticus (spherical, non-motile) in shake-flask cultures at various rotating speeds. For all the strains, a higher rotating speed always resulted in a shorter lag phase, indicating that the strains showed a superior adaptability under higher hydrodynamic conditions. The maximum specific growth rate of M. oxydans, A. faecalis, and S. haemolyticus increased rapidly with the increase of energy dissipation rate till the highest value of 0.386, 0.240, and 0.323 1/h and then decreased as the rotating speed further increased. The phenol biodegradation rate was also dependent on rotating speed, and the trends were consistent with the growth rate variations. A predictive model similar to Haldane model was proposed and was fitted well (R²?>?0.913) with bacterial growth under different hydrodynamic conditions. According to the predictive model, the optimum hydrodynamic conditions for the growth of M. oxydans, A. faecalis, and S. haemolyticus were 3.099, 2.197, and 2.289 m²/s³, respectively. The results suggested that non-motile and rod-shaped bacteria were more dependent on hydrodynamic conditions than motile and spherical ones, which could be attributed to the discrepancies in bacterial morphology and motility. The results provide a better understanding on bacterial responses to various hydrodynamic conditions and could be further applied in the bioremediation of contaminated water.     

The foliar spray of <Emphasis Type="Italic">Rhodopseudomonas palustris</Emphasis> grown under <Emphasis Type="Italic">Stevia</Emphasis> residue extract promotes plant growth via changing soil microbial community

Purpose

The extract of Stevia residue is an ideal substitute for cultivation of the purple nonsulfur bacterium, like Rhodopseudomonas palustris (R. palustris). But the influence of R. palustris grown under residue extract on its downstream application is still not well-characterized. The objective of this study was to assess the effect of foliar spray of R. palustris grown under Stevia residue extract on the plant growth and soil microbial properties.

Materials and methods

A pot experiment was carried out under the greenhouse condition, consisting of four treatments varying in the sprayed substances: sterilized water (control), R. palustris grown under the chemical medium supplemented with L-tryptophan (SyT), R. palustris grown under Stevia residue extract supplemented with L-tryptophan (ExT), and R. palustris grown under Stevia residue extract supplemented with NH₄Cl (ExT). The net photosynthesis rate of the uppermost leaves was measured with a portable photosynthesis system. Soil microbial activity was analyzed by microcalorimetry. Soil bacterial community components were determined by real-time quantitative PCR (qPCR) and high-throughput sequencing techniques.

Results and discussion

Compared with SyT, the R. palustris grown under Stevia residue extract not only improved the plant biomass and the net photosynthetic rate to a large extent, but also increased soil microbial metabolic activity and altered community compositions as well. The treatments receiving R. palustris, especially ExT and ExN, increased the relative abundances of some functional guilds involved in C turnover and nutrient cycling in soil, including Acidobacteria, Actinobacteria, Proteobacteria, Gemmatimonadaetes, Nitrospirae, and Planctomycetes.

Conclusions

R. palustris grown under the Stevia residue extract showed advantages over that under the chemical medium on both plant growth and soil microbial properties. One of the possible reasons could result from the increases in microbial activity and several bacterial keystone guilds involved into C and nutrient cycling, both of which potentially contribute to the improved plant growth. The results would be conducive to the downstream application of R. palustris in an economical way.

     

Bioremediation of Hexachlorocyclohexane-Contaminated Soil by the New <Emphasis Type="Italic">Rhodococcus wratislaviensis</Emphasis> Strain Ch628

Currently, there are large areas of soils contaminated with hexachlorocyclohexane (HCH) isomers which are included in the group of persistent organic pollutants. For the bioremediation of such soils, a new HCH-degrading Rhodococcus wratislaviensis strain Ch628 was isolated from long-term organochlorine contaminated soils. The strain Ch628 was able to degrade 32.3% γ-hexachlorocyclohexane (γ-HCH/lindane), 25.2% hexachlorobenzene, and 100% chlorobenzene in resting cell conditions. The strain Ch628 was bioaugmented in chronically HCH-contaminated soil. The results showed that the bioaugmentation of contaminated soil with the strain Ch628 led to HCH degradation. In the bioaugmented system, the efficiency of HCH removal at the initial concentration of about 238.7 ± 4.9 mg kg^?1 soil was 44.8%, while the system with indigenous microflora (without R. wratislaviensis strain Ch628) and the system with abiotic control removed 33.3 and 16.4% of this compound during the same period, respectively. Strain Ch628 could effectively degrade α-, β-, and γ-isomers of HCH (77.1, 100, and 100%, respectively) and heptachlorocyclohexane (69.9%) in the model soil systems. Moreover, the bioaugmentation with the strain Ch628 led to degradation of tri-, tetra-, and penta-chlorobenzenes, which are of HCH degradation metabolites. For the first time, it was found that the bioaugmentation with the bacterial strain Rhodococcus wratislaviensis Сh628 led to a significant reduction of the toxicity of the HCH-contaminated soil for the test organisms, such as Chlorella vulgaris Beijer and Daphnia magna Straus.     

Catalase and Phosphatase Activities During Hydrocarbon Removal from Oil-Contaminated Soil Amended with Agro-Industrial By-products and Macronutrients

Microbiological activities are essential in the bioremediation of polluted soils. The enzymatic activities of microorganisms are usually used as a biological indicator of soil health. The aim of this work was to observe the catalase, acid phosphatase (AcP), and alkaline phosphatase (AlP) activities in soil that was amended with agro-industrial by-products and macronutrients during the process of total petroleum hydrocarbon (TPH) removal. To this end, microcosm tests were performed with soil and agro-industrial by-products ratios of 100:2:2, for soil:sugarcane bagasse pith:filter cake mud (SSF); 100:2, for both soil:sugarcane bagasse pith (SS); and for soil filter cake mud (SF). The macronutrients—carbon, nitrogen, and phosphorus—in the experimental treatments were adjusted to 100:10:1 with a solution of NH₄NO₃ and K₂HPO₄. The best TPH removal (51.4%) was obtained with SSF at 15 days. In addition, a significant correlation was observed between TPH removal and AlP as well as AcP (r = 0.74, p < 0.0001; r = 0.70, p < 0.0107, respectively). Fungi growth was also correlated with both AlP (r = 0.97, p < 0.0001) and AcP (r = 0.95, p < 0.0001) activities. Besides, bacterial and fungi growth showed a correlation with TPH (r = 0.86, p < 0.001; r = 0.77, p < 0.0034, respectively). It could be said that the agro-industrial by-products and macronutrients contributed to pollutant removal from the oil-polluted soil at relatively short amount of time. In addition, the enzymatic activities were increased after the treatment; in this study, the high sensitivity enzyme was AlP, and it could be used as an indirect indicator of oil pollutant removal.     

Phytostabilization Ability of <Emphasis Type="Italic">Baccharis linearis</Emphasis> and Its Relation to Properties of a Tailings-Derived Technosol

Spontaneous colonization of mine tailing dams by plants is a potential tool for phytostabilization of such reservoirs. However, the physical and chemical properties of each mine tailings deposit determine the success of natural plant establishment. The plant Baccharis linearis is the main native nanophanerophyte species (evergreen sclerophyllous shrub) that naturally colonizes abandoned copper tailings dams in arid to semiarid north-central Chile. This study compare growth of B. linearis against the physical and chemical properties of a Technosol derived from copper mine tailings. Five sites inside the deposit were selected based on B. linearis vegetation density (VD), at two soil sampling depths under the canopy of adult individuals. Physical and chemical properties of tailings samples and nutrient concentrations in tailings and plants were each determined. Some morphological features of the plants (roots and aerial parts) were also quantified. There were significant differences in soil available water capacity (AW) and relative density (Rd) at different VD. Sites with low AW and high Rd had lower nutrient concentrations and higher Zn content in tailings, decreased infection by arbuscular mycorrhizal fungi, and increased fine root abundance and root hair length in individual plants. In contrast, higher AW, which was positively correlated with fine particles and organic matter content, had a positive effect on vegetation coverage, increased N and P contents in tailings, and increased N contents in leaf tissues, even when available N and P levels in tailings were low. Multiple constraints, such as low AW, N, P, and B contents and high Zn concentrations in the tailings restricted vegetation coverage, but no phenotypic differences were observed between individuals. Thus, in order to promote dense coverage by B. linearis, water retention in these tailings must be improved by increasing colloidal particles (organic and/or inorganic) contents, which have a positive effect on colonization by this species.     

Assessment of <Emphasis Type="Italic">Koelreuteria paniculata</Emphasis> Seedling for Phytroremediation of Pyrene-Contaminated Soils

Phytoremediation is a cost-effective and environmentally friendly technology using plants for the cleanup of both inorganic and organic contaminated sites. In this study, a pot culture experiment has been conducted for 180 days in a greenhouse to examine the capability of Koelreuteria paniculata on pyrene (Pyr) dissipation in contaminated soil. Three treatments were employed and they were: (1) polluted soil with K. paniculata fine roots addition (T1), (2) polluted soil with planted seedlings (T2), and (3) polluted soil (C). Results showed Pyr concentration in soils was reduced by 21.4, 36.2, and 86.4% by natural losses, fine roots addition, and planted K. paniculata treatments, respectively, meaning plants substantially enhanced the dissipation of Pyr from soil. Cultivated K. paniculata seedlings significantly increased soil total nitrogen (TN), total organic carbon, dissolved organic carbon (DOC), and microbial biomass carbon, but not total phosphorus, when compared to the control. The removal efficiency of Pyr was lower in the adding of fine roots treatment than in the planted K. paniculata treatment. The principal component analysis indicated the promotional dissipation of Pyr in soil by planted K. paniculata was likely attributed to increased microbial quantity and activity, DOC, and TN content in the rhizosphere. Our results suggest that K. paniculata is a suitable plant species used in phytoremediation for Pyr-contaminated soils and the efficiency on the dissipation of Pyr is considerably enhanced using living plants than adding dead organic matters. The study provided a reference for the application of K. paniculata in the remediation of Pyr-contaminated soil.     

Optimization of <Emphasis Type="Italic">o</Emphasis>-Chlorophenol Biodegradation by Combined Mycelial Pellets Using Response Surface Methodology

In the present study, the immobilizing fermentation characteristics and o-chlorophenol biodegradation of Rhodopseudomonas palustris using mycelial pellets as a biomass carrier were investigated. To improve the o-chlorophenol degradation efficiency of the combined mycelial pellets, eight cultivation variables including glucose concentration, yeast extract concentration, spore inoculum size, pH, and agitation speed were optimized with an integrated strategy involving a combination of statistical designs. First, Plackett-Burman experiments identified glucose, yeast extract, and spore inoculum size as three statistically significant factors important for o-chlorophenol removal. Then, the steepest ascent method was used to access the optimal region of these significant factors. Finally, response surface methodology by Box-Behnken optimization was used to examine the mutual interactions among these three variables to determine their optimal levels. The ideal culture conditions for maximum o-chlorophenol removal according to a second-order polynomial model were as follows: 15.60 g/L glucose, 3.09 g/L yeast extract, and 9% (v/v) spore inoculum size, resulting in an expected o-chlorophenol removal rate of 92.60% with an o-chlorophenol initial concentration of 50 mg/L and 96-h culture time. The correlation coefficient (R ² = 0.9933) indicated excellent agreement between the experimental and predicted values, whereas a fair association was observed between the predicted model values and those obtained from subsequent experimentation at the optimized conditions.     

Estimation of natural outcrossing rate and genetic diversity in Lima bean (<Emphasis Type="Italic">Phaseolus lunatus</Emphasis> L. var. <Emphasis Type="Italic">lunatus</Emphasis>) from Brazil using SSR markers: implications for conservation and breeding

Lima bean (Phaseolus lunatus L.) is an important food source in Brazil, especially in the northeast region, where its production and consumption are high. The goals of the present study were to estimate natural outcrossing rates and genetic diversity levels of Lima bean from Brazil, using ten microsatellite loci to obtain information for their conservation and breeding. Fourteen accessions were selected from an experiment in field with open-pollinated and with the presence of pollinating insects. Twelve seeds of each of the 14 selected accessions were grown in screenhouse for tissue harvest and DNA extraction. The multilocus model was used to determine the reproductive system. The outcrossing rate was 38.1 % (t_m = 0.381; t_s = 0.078), and the results indicated a mixed mating system with a predominance of selfing (1 ? t_m = 61.9 %). The biparental inbreeding rate was high (t _m  ? t _s  = 0.303) and the multilocus correlated paternity was quite high (r _p(m) = 0.889), indicating that the progeny was mostly composed of full sibs. The average effective number of pollen donors per maternal plant (N _ep ) was low (1.12), and the fixation index for maternal genotypes (F _m ) was 0.945, indicating that most genitors resulted from inbreeding. The studied families presented considerable genetic variability: A = 6.10;  %P = 30; H _e  = 0.60 and H _o  = 0.077. Total diversity was high (H _T = 0.596), and a portion was distributed within families (H _S = 0.058). In addition, diversity was higher between families (D _ST = 0.538), and genetic differentiation was high (G _ST = 0.902). The results presented here can be used in the implementation of Lima bean conservation and breeding programs in Brazil.     

<Emphasis Type="Italic">p</Emphasis>-Coumaric can alter the composition of cucumber rhizosphere microbial communities and induce negative plant-microbial interactions

Phenolics from root exudates or decaying residues are usually referred as autotoxins of several plant species. However, how phenolics affect soil microbial communities and their functional significances are poorly understood. Rhizosphere bacterial and fungal communities from cucumber (Cucumis sativus L.) seedlings treated with p-coumaric acid, an autotoxin of cucumber, were analyzed by high-throughput sequencing of 16S rRNA gene and internal transcribed spacer amplicons. Then, feedback effects of the rhizosphere biota on cucumber seedlings were evaluated by inoculating non-sterilized and sterilized rhizosphere soils to sterilized background soils. p-Coumaric acid decreased the bacterial diversity of rhizosphere but increased fungal diversity and altered the compositions of both the bacterial and fungal communities. p-Coumaric acid increased the relative abundances of microbial taxa with phenol-degrading capability (such as Chaetomium, Humicola, and Mortierella spp.) and microbial taxa which contained plant pathogens (such as Fusarium spp.). However, p-coumaric acid inhibited the relative abundances of Lysobacter, Haliangium, and Gymnoascus spp., whose species can have pathogen-antagonistic and/or plant-growth-promoting effects. The positive effect of cucumber rhizosphere microbiota on cucumber seedling growth was reduced by p-coumaric acid. Overall, our results showed that, besides its direct phytotoxicity, p-coumaric acid can inhibit cucumber seedling growth through generating negative plant-soil microbial interactions.     

Effect of the Simultaneous Action of Zinc and Chromium on <Emphasis Type="Italic">Arthrobacter</Emphasis> spp.

Bacteria are regarded as the most effective in the detoxification of heavy metals, being environmental compatible. Metalloresistant bacteria are usually found in nature in highly contaminated environment where they interact with a combination of several toxic metals. For the present research, Arthrobacter oxydans and Arthrobacter globiformis have been isolated from the soil samples of the most polluted regions of Georgia, rich with manganese and iron, and contain co-produced toxic metals such as Cr, V, Zn, Ni, Pb, and Mo. We have studied the effects of the metals with different valence/charge on the metalloresistant Arthrobacter spp., the divalent cation—Zn(II) and the hexavalent anion—Cr(VI). The permanent presence of a nontoxic concentration of zinc alone or zinc together with the subtoxic concentration of chromium at the growth of A. oxydans and A. globiformis as batch culture causes the activation of the zinc primary uptake system transporters from the ZIP family (Zrt1). Chromium does not affect the process. The studied Arthrobacter spp. differ by the character of the activation of the antioxidant defense system. Chromium and zinc concomitant action causes the strongest oxidative stress in the case of A. globiformis that is demonstrated by the increased activity of superoxide dismutase (SOD) and catalase. In the case of A. oxydans, the zinc separate action, and the joint action of zinc and chromium decreases the activity of SOD and catalase. The antioxidant system is active in A. globiformis at the prolonged action of metals (96 h), whereas the cells of A. oxyidans activate the other defense mechanisms to survive.     

Potential of Pyrene Removal from Urban Environments by the Activities of Bacteria and Biosurfactant on Ornamental Plant Leaves

Pyrene is a dominant PAH in urban environments. It can combine with airborne particulates and accumulate on plant leaves. To investigate pyrene’s biodegradation potential, this study initially monitored the abundance of airborne and phyllosphere bacteria. The number of airborne pyrene-degrading bacteria ranged from 22 to 152 CFU m^?3 air, and more bacteria were found in the proximity of the ornamental plant swath than along the roadside. Pyrene-degrading bacteria averaged 5 × 10⁴ CFU g^?1 on the leaves of all tested plant species and accounted for approximately 7% of the total population. Four pyrene-degrading bacteria were isolated from I. coccinea to use as model phyllosphere bacteria. To increase the bioavailability of pyrene, a lipopeptide biosurfactant was applied. Kocuria sp. IC3 showed the highest pyrene degradation in the medium containing biosurfactant. The removal of deposited pyrene at 30 μg g^?1 leaf was monitored in a glass chamber containing I. coccinea twigs. After 14 days, leaves containing both Kocuria sp. IC3 and 0.1× CMC biosurfactant showed 100% pyrene removal with the most abundant bacteria. The system with biosurfactant alone also enhanced the activities of phyllosphere bacteria with 94% pyrene removal. Consequently, the bioremediation of deposited pyrene could be achieved by spraying biosurfactant on ornamental shrubs.     

Effects of biochar on Cd and Pb mobility and microbial community composition in a calcareous soil planted with tobacco

An experiment was conducted with tobacco (Nicotiana tabacum L.) grown in a Cd- and Pb-contaminated calcareous soil amended with 0.0, 1.0, 2.5, and 5.0% (w/w) tobacco stalk biochar (BC). The BC amendment significantly increased organic matter, total C, N, P, and K contents of soil, and the C/N ratio. Bioavailable metal concentrations (DTPA extraction) decreased by increasing BC application rate. The 5.0% BC amendment significantly decreased the DTPA-extractable Cd and Pb by 10.4 and 13.6%, respectively. Correspondingly, the bioaccumulation and translocation factors of Cd and Pb also decreased by increasing the BC addition rates and this indicated that BC inhibited the uptake and transfer of both Cd and Pb by tobacco plants. Moreover, high-throughput sequencing revealed that BC increased Chao1 richness, Shannon’s diversity and Simpson’s diversity of bacterial communities of soil. The relative abundance and genera composition of Adhaeribacter, Rhodoplanes, Pseudoxanthomonas, and Candidatus Xiphinematobacter increased under BC treatments, while those of Kaistobacter, Lacibacter, and Pirellula decreased. Overall, BC increased soil nutrients (C, N, P, and K contents), enhanced bacterial diversity indexes and richness, and changed the bacterial community composition, which may all have contributed to reduce the mobility and bioavailability of both Cd and Pb in a calcareous soil.