Cell wall canals formed upon growth of Candida maltosa in the presence of hexadecane are associated with polyphosphates.

Canals are supramolecular complexes observed in the cell wall of Candida maltosa grown in the presence of hexadecane as a sole carbon source. Such structures were not observed in glucose-grown cells. Microscopic observations of cells stained with diaminobenzidine revealed the presence of oxidative enzymes in the canals. 4΄,6΄-diamino-2-phenylindole staining revealed that a substantial part of cellular polyphosphate was present in the cell wall of cells grown on hexadecane in condition of phosphate limitation. The content and chain length of polyphosphates were higher in hexadecane-grown cells than in glucose grown ones. The treatment of cells with yeast polyphosphatase PPX1 resulted in the decrease of the canal size. These data clearly indicated that polyphosphates are constituents of canals; they might play an important role in the canal structure and functioning.

Synergistic use of biochar, compost and plant growth-promoting rhizobacteria for enhancing cucumber growth under water deficit conditions.

BACKGROUND: Limited information is available about the effectiveness of biochar with plant growth-promoting rhizobacteria (PGPR) and compost. A greenhouse study was conducted to evaluate the effect of biochar in combination with compost and PGPR (Pseudomonas fluorescens) for alleviating water deficit stress. Both inoculated and un-inoculated cucumber seeds were sown in soil treated with biochar, compost and biochar + compost. Three water levels - field capacity (D0), 75% field capacity (D1) and 50% field capacity (D2) - were maintained. RESULTS: The results showed that water deficit stress significantly suppressed the growth of cucumber; however, synergistic use of biochar, compost and PGPR mitigated the negative impact of stress. At D2, the synergistic use of biochar, compost and PGPR caused significant increases in shoot length, shoot biomass, root length and root biomass, which were respectively 88, 77, 89 and 74% more than in the un-inoculated control. Significant improvements in chlorophyll and relative water contents as well as reduction in leaf electrolyte leakage demonstrated the effectiveness of this approach. Moreover, the highest population of P. fluorescens was observed where biochar and compost were applied together. CONCLUSION: These results suggest that application of biochar with PGPR and/or compost could be an effective strategy for enhancing plant growth under stress. © 2017 Society of Chemical Industry.

Modifications of the cell wall of yeasts grown on hexadecane and under starvation conditions.

Electron-microscopic examinations have demonstrated local modifications in the cell wall of the yeast Candida maltosa grown on hexadecane. In our earlier studies, these modified sites, observed in other yeasts grown on oil hydrocarbons, were conventionally called 'canals'. The biochemical and cytochemical studies of C. maltosa have revealed a correlation between the formation of 'canals' and decrease in the amount of cell wall polysaccharides, glucan and mannan. The ultrathin sections and surface replicas have shown that the 'canals' are destroyed by pronase, thus indicating that a significant proportion of their content is represented by proteins. This finding was compatible with our earlier data on the localization of oxidative enzymes in 'canals' and possible participation of the 'canals' in the primary oxidation of hydrocarbons. A completely unexpected and intriguing phenomenon has been the appearance of 'canals' in the yeast C. maltosa under starvation conditions. Unlike the yeasts grown on hexadecane, mannan almost disappears in starving cells, while the quantity of glucan first decreases and then is restored to its initial level. The role of 'canals' in starving cells is as yet unclear; it is assumed that they acquire exoenzymes involved in the utilization of products of cell lysis in the starving population. In the future, 'canals' of starving cells will be studied in connection with their possible participation in apoptosis.

Detoxification of azo dyes by bacterial oxidoreductase enzymes.

Azo dyes and their intermediate degradation products are common contaminants of soil and groundwater in developing countries where textile and leather dye products are produced. The toxicity of azo dyes is primarily associated with their molecular structure, substitution groups and reactivity. To avoid contamination of natural resources and to minimize risk to human health, this wastewater requires treatment in an environmentally safe manner. This manuscript critically reviews biological treatment systems and the role of bacterial reductive and oxidative enzymes/processes in the bioremediation of dye-polluted wastewaters. Many studies have shown that a variety of culturable bacteria have efficient enzymatic systems that can carry out complete mineralization of dye chemicals and their metabolites (aromatic compounds) over a wide range of environmental conditions. Complete mineralization of azo dyes generally involves a two-step process requiring initial anaerobic treatment for decolorization, followed by an oxidative process that results in degradation of the toxic intermediates that are formed during the first step. Molecular studies have revealed that the first reductive process can be carried out by two classes of enzymes involving flavin-dependent and flavin-free azoreductases under anaerobic or low oxygen conditions. The second step that is carried out by oxidative enzymes that primarily involves broad specificity peroxidases, laccases and tyrosinases. This review focuses, in particular, on the characterization of these enzymes with respect to their enzyme kinetics and the environmental conditions that are necessary for bioreactor systems to treat azo dyes contained in wastewater.

Relationship between in vitro characterization and comparative efficacy of plant growth-promoting rhizobacteria for improving cucumber salt tolerance.

Phosphate solubilization, 1-aminocyclopropane-1-carboxylic acid (ACC)-deaminase activity and production of siderophores and indole acetic acid (IAA) are well-known traits of plant growth-promoting rhizobacteria (PGPR). Here we investigated the expression of these traits as affected by salinity for three PGPR strains (Pseudomonas fluorescens, Bacillus megaterium and Variovorax paradoxus) at two salinity levels [2 and 5 % NaCl (w/v)]. Among the three strains, growth of B. megaterium was the least affected by high salinity. However, P. fluorescens was the best strain for maintaining ACC-deaminase activity, siderophore and IAA production under stressed conditions. V. paradoxus was the least tolerant to salts and had minimal growth and low PGPR trait expression under salt stress. Results of experiment examining the impact of bacterial inoculation on cucumber growth at three salinity levels [1 (normal), 7 and 10 dS m(-1)] revealed that P. fluorescens also had good rhizosphere competence and was the most effective for alleviating the negative impacts of salinity on cucumber growth. The results suggest that in addition to screening the PGPR regarding their effect on growth under salinity, PGPR trait expression is also an important aspect that may be useful for selecting the most promising PGPR bacterial strains for improving plant tolerance to salinity stress.

Yeast extract promotes decolorization of azo dyes by stimulating azoreductase activity in Shewanella sp. strain IFN4.

Biological treatment of azo dyes commonly requires a combined anaerobic-aerobic process in which initial decolorization is achieved by reductive cleavage of azo bonds on the parent molecule. The present study was conducted to examine the relative importance of co-substrates for driving reductive decolorization of azo dyes by Shewanella sp. strain IFN4 using whole cells and enzyme assays. Results showed that the dye decolorization by strain IFN4 was faster in medium containing 1gL(-1) yeast extract (YE) as compared to nine other co-substrates. Moreover, only YE stimulated azoreductase activity (increased from 1.32 to 4.19U/mg protein). Increasing the level of YE up to 8gL(-)(1) resulted into 81% decolorization of the dye in 1h along with an increase in azoreductase activity up to 6.16U/mg protein. Among the components of YE, only riboflavin stimulated the decolorization process as well as enzyme activity. Moreover, strain IFN4 demonstrated flavin reductase activity, and a significant correlation (r(2)=0.98) between flavin reduction and dye reduction by this strain emphasized the involvement of flavin compounds in the decolorization process. The results of this study show that YE serves both as a source of reducing equivalents and an electron shuttle for catalyzing dye reduction.

The stability of textile azo dyes in soil and their impact on microbial phospholipid fatty acid profiles.

The aim of this study was to examine the stability of structurally different azo dyes in soil and their impact on the microbial community composition by analyzing phospholipid fatty acid (PLFA) profiles. Sterile and non-sterile soils were amended with three azo dyes, including: Direct Red 81, Reactive Black 5 and Acid Yellow 19 at 160mgkg(-1) soil. The results showed that the azo dyes were quite stable and that large amounts of these dyes ranging from 17.3% to 87.5% were recoverable from the sterile and non-sterile soils after 14 days. The maximum amount of dye was recovered in the case of Direct Red 81. PLFA analysis showed that the azo dyes had a significant effect on microbial community structure. PLFA concentrations representing Gram-negative bacteria in dye-amended soil were substantially less as compared to the PLFA concentration of Gram-positive bacteria. Acid Yellow 19 dye had almost similar effects on the PLFA concentrations representing bacteria and fungi. In contrast, Reactive Black 5 had a greater negative effect on fungal PLFA than that on bacterial PLFA, while the opposite was observed in the case of Direct Red 81. To our knowledge, this is the first study reporting the stability of textile azo dyes in soil and their effects on soil microbial community composition.

Decolorization of Reactive Black-5 by Shewanella sp. in the Presence of Metal Ions and Salts.

In this study, effect of various metal ions and salts on biodecolorization of Reactive black-5, azoreductase activity, and growth of Shewanella sp. strain IFN4 was evaluated. Among the tested metals, Cr²âº, Pb(²âº, Ni²âº, Fe²âº, and Mn²âº did not inhibit the biodecolorization of reactive black-5, azoreductase activity and bacterial growth. Three metals (Cu²âº, Zn²âº, and Co²âº) delayed the decolorization process without completely inhibiting the reaction and also suppressed the bacterial growth. However, no dye decolorization was observed in the presence of Cd²âº (10 mg L⁻¹). Furthermore, bacterium decolorized the dye at high concentration (15 mg L⁻¹) of mixed metal ions. Strain IFN4 was also able to decolorize the dye at 50 g NaCl L⁻¹ and 60 g Na2SO4 L⁻¹. NaCl was found to be more inhibitory to bacterial growth than Na2SO4and the reverse was observed for azoreductase activity. These findings suggest that strain IFN4 could be used in designing a bioreactor for the treatment of textile effluent.

Study of phenanthrene utilizing bacterial consortia associated with cowpea (Vigna unguiculata) root nodules.

Many legumes have been selected as model plants to degrade organic contaminants with their special associated rhizosphere microbes in soil. However, the function of root nodules during microbe-assisted phytoremediation is not clear. A pot study was conducted to examine phenanthrene (PHE) utilizing bacteria associated with root nodules and the effects of cowpea root nodules on phytoremediation in two different types of soils (freshly contaminated soil and aged contaminated soil). Cowpea nodules in freshly-contaminated soil showed less damage in comparison to the aged-contaminated soil, both morphologically and ultra-structurally by scanning electron microscopy. The study of polycyclic aromatic hydrocarbon (PAH) attenuation conducted by high performance liquid chromatography revealed that more PAH was eliminated from liquid culture around nodulated roots than nodule-free roots. PAH sublimation and denaturation gradient gel electrophoresis were applied to analyze the capability and diversity of PAH degrading bacteria from the following four parts of rhizo-microzone: bulk soil, root surface, nodule surface and nodule inside. The results indicated that the surface and inside of cowpea root nodules were colonized with bacterial consortia that utilized PHE. Our results demonstrated that root nodules not only fixed nitrogen, but also enriched PAH-utilizing microorganisms both inside and outside of the nodules. Legume nodules may have biotechnological values for PAH degradation.

Effects of cowpea (Vigna unguiculata) root mucilage on microbial community response and capacity for phenanthrene remediation.

Biodegradation of polycyclic aromatic hydrocarbons (PAHs) is normally limited by their low solubility and poor bioavailability. Prior research suggests that biosurfactants are synthesized as intermediates during the production of mucilage at the root tip. To date the effects of mucilage on PAH degradation and microbial community response have not been directly examined. To address this question, our research compared 3 cowpea breeding lines (Vigna unguiculata) that differed in mucilage production for their effects on phenanthrene (PHE) degradation in soil. The High Performance Liquid Chromatography results indicated that the highest PHE degradation rate was achieved in soils planted with mucilage producing cowpea line C1, inoculated with Bradyrhizobium, leading to 91.6% PHE disappearance in 5 weeks. In root printing tests, strings treated with mucilage and bacteria produced larger clearing zones than those produced on mucilage treated strings with no bacteria or bacteria inoculated strings. Experiments with 14C-PHE and purified mucilage in soil slurry confirmed that the root mucilage significantly enhanced PHE mineralization (82.7%), which is 12% more than the control treatment without mucilage. The profiles of the PHE degraders generated by Denaturing gradient gel electrophoresis suggested that cowpea C1, producing a high amount of root mucilage, selectively enriched the PHE degrading bacteria population in rhizosphere. These findings indicate that root mucilage may play a significant role in enhancing PHE degradation and suggests that differences in mucilage production may be an important criterion for selection of the best plant species for use in phytoremediation of PAH contaminated soils.

Bacterial community assemblages associated with the phyllosphere, dermosphere, and rhizosphere of tree species of the Atlantic forest are host taxon dependent.

Bacterial communities associated with tree canopies have been shown to be specific to their plant hosts, suggesting that plant species-specific traits may drive the selection of microbial species that comprise their microbiomes. To further examine the degree to which the plant taxa drive the assemblage of bacterial communities in specific plant microenvironments, we evaluated bacterial community structures associated with the phyllosphere, dermosphere, and rhizosphere of seven tree species representing three orders, four families and four genera of plants from a pristine Dense Ombrophilous Atlantic forest in Brazil, using a combination of PCR-DGGE of 16S rRNA genes and clone library sequencing. Results indicated that each plant species selected for distinct bacterial communities in the phyllosphere, dermosphere, and rhizosphere, and that the bacterial community structures are significantly related to the plant taxa, at the species, family, and order levels. Further characterization of the bacterial communities of the phyllosphere and dermosphere of the tree species showed that they were inhabited predominantly by species of Gammaproteobacteria, mostly related to Pseudomonas. In contrast, the rhizosphere bacterial communities showed greater species richness and evenness, and higher frequencies of Alphaproteobacteria and Acidobacteria Gp1. With individual tree species each selecting for their specific microbiomes, these findings greatly increase our estimates of the bacterial species richness in tropical forests and provoke questions concerning the ecological functions of the microbial communities that exist on different plant parts.

Draft genome sequence of Rahnella aquatilis strain HX2, a plant growth-promoting rhizobacterium isolated from vineyard soil in Beijing, China.

Rahnella aquatilis strain HX2 is a plant growth-promoting, disease-suppressive rhizobacterium that was isolated from a vineyard soil in Beijing, China. Here, we report the genome sequence of this strain, which provides a valuable resource for future research examining the mechanisms of traits associated with plant growth promotion and biocontrol.

Plant growth-promoting rhizobacteria associated with ancient clones of creosote bush (Larrea tridentata).

Plant growth-promoting rhizobacteria (PGPR) are common components of the rhizosphere, but their role in adaptation of plants to extreme environments is not yet understood. Here, we examined rhizobacteria associated with ancient clones of Larrea tridentata in the Mohave desert, including the 11,700-year-old King Clone, which is oldest known specimen of this species. Analysis of unculturable and culturable bacterial community by PCR-DGGE revealed taxa that have previously been described on agricultural plants. These taxa included species of Proteobacteria, Bacteroidetes, and Firmicutes that commonly carry traits associated with plant growth promotion, including genes encoding aminocyclopropane carboxylate deaminase and ß-propeller phytase. The PGPR activities of three representative isolates from L. tridentata were further confirmed using cucumber plants to screen for plant growth promotion. This study provides an intriguing first view of the mutualistic bacteria that are associated with some of the world's oldest living plants and suggests that PGPR likely contribute to the adaptation of L. tridentata and other plant species to harsh environmental conditions in desert habitats.

Persistence of Escherichia coli O157:H7 in major leafy green producing soils.

Persistence of Escherichia coli O157:H7 in 32 (16 organically managed and 16 conventionally managed) soils from California (CA) and Arizona (AZ) was investigated. Results showed that the longest survival (ttd, time needed to reach detection limit, 100 CFU g(-1) dry soil) of E. coli O157:H7 was observed in the soils from Salinas Valley, CA and in organically managed soils from AZ. Detrended correspondence analysis revealed that the survival profiles in organically managed soils in Yuma, AZ were different from the ones in conventionally managed soils from the same site. Principal component analysis and stepwise regression analysis showed that E. coli O157:H7 survival in soils was negatively correlated with salinity (EC) (P < 0.001), while positively correlated with assimilable organic carbon (AOC) and total nitrogen (TN) (P < 0.01). Pearson correlation analysis revealed that a greater ttd was associated with a larger δ (time needed for first decimal reduction in E. coli population). EC was negatively correlated and TN was positively correlated (P < 0.05) with δ, suggesting that EC and TN likely have a direct impact on ttd. On the other hand, AOC showed a close correlation with p (the shape parameter) that was not directly related to ttd, indicating that AOC might have an indirect effect in the overall survival of E. coli O157:H7 in soils. Our data showed that AOC and EC significantly affected the survival of E. coli O157:H7 in leafy green producing soils and the development of good agricultural practices (manure/composting/irrigation water source management) in the preharvest environment must be followed to minimize foodborne bacterial contamination on fresh produce.

Isolation of biosurfactant-producing bacteria from the Rancho La Brea Tar Pits.

This research was conducted to identify culturable surfactant-producing bacterial species that inhabit the 40,000-year-old natural asphalt seep at the Rancho La Brea Tar Pits in Los Angeles, CA. Using phenanthrene, monocyclic aromatic hydrocarbons, and tryptic soy broth as growth substrates, culturable bacteria from the tar pits yielded ten isolates, of which three species of gamma-proteobacteria produced biosurfactants that accumulated in spent culture medium. Partially purified biosurfactants produced by these strains lowered the surface tension of water from 70 to 35-55 mN/m and two of the biosurfactants produced 'dark halos' with the atomized oil assay, a phenomenon previously observed only with synthetic surfactants. Key findings include the isolation of culturable biosurfactant-producing bacteria that comprise a relatively small fraction of the petroleum-degrading community in the asphalt.

Nested PCR bias: a case study of Pseudomonas spp. in soil microcosms.

Nested PCR methods combined with denaturing gradient gel electrophoresis (DGGE) are widely used for the detection of low copy number species or for the analysis of group-specific community profiles. With an appropriate number of PCR cycles during the first round of amplification, initial differences in the copy numbers of different DNA fragments that are targeted can be maintained during the second round without significant bias. However, if an excessive number of cycles in used in the first round, relative differences in the copy numbers of the targeted sequences can be obscured. Here we demonstrate the effect of "nested PCR bias" in a case study with PCR-DGGE of 16S rRNA gene sequences targeting Pseudomonas spp. following exposure of soil to naphthalene vapors. Our results demonstrate artifacts caused by nested PCR bias can be substantially minimized by calibrating the number of first round PCR cycles, thereby preserving the ability to obtain semiquantitative data for evaluating changes in gene copy numbers over time.

Biodegradation potential of pure and mixed bacterial cultures for removal of 4-nitroaniline from textile dye wastewater.

Environmentally toxic aromatic amines including nitroanilines are commonly generated in dye contaminated wastewater in which azo dyes undergo degradation under anaerobic conditions. The aim of this study was to develop a process for biological treatment of 4-nitroaniline. Three bacteria identified as Acinetobacter sp., Citrobacter freundii and Klebsiella oxytoca were isolated from enrichment cultures of activated sludge on 4-nitroaniline, after which the isolates and the mixed culture were studied to determine optimal conditions for biodegradation. HPLC analyses showed the mixed culture was capable of complete removal of 100micromol/L of 4-nitroaniline within 72h under aerobic conditions. There was an inverse linear relationship (R(2)=0.96) between the rate of degradation (V) and 4-nitraoaniline concentrations [S] over 100-1000micromol/L. The bacterial culture was also capable of decolorizing structurally different azo dyes (Acid Red-88, Reactive Black-5, Direct Red-81, and Disperse Orange-3) and also degraded nitrobenzene. Our findings show that enrichment cultures from activated sludge can be effective for the removal of dyes and their toxic intermediates, and that treatment may best be accomplished using an anaerobic-aerobic process.

Biochar effects on uptake of cadmium and lead by wheat in relation to annual precipitation: a 3-year field study.

Biochar has been widely studied for its ability to reduce plant uptake of heavy metals by lowering metal bioavailabilities through adsorption and pH-driven fixation reactions. However, the long-term effect of biochar on heavy metal bioavailabilities in alkaline soils under natural redox condition is rarely studied. Here, we report a study examining the effects of biochar on bioavailability and partitioning of cadmium (Cd) and lead (Pb) among different soil fractions over 3 years in a field study with wheat (Triticum aestivum L.). Plots were established on two similar soils having low and high levels of contamination, both of which were amended in the first year with wheat straw biochar at 0, 20, and 40 t ha-1. Precipitation patterns varied greatly over the study period, with 2014 having record drought, which was followed by 2 years having extreme flooding events. Results showed a significant increase in grain yield and reductions in Cd and Pb concentrations in wheat grain in the biochar-amended soils in 2014. In contrast, bioavailable (exchangeable) heavy metal concentrations and plant uptake of Cd and Pb were significantly higher in the subsequent very wet years in 2015 and 2016, where the effects of biochar were much more variable and had an overall lesser effect on reducing heavy metal uptake. The results suggest that fluctuations in soil pH and redox caused by periodic drought and flood cycles strongly drive metal cycling through mobilization and immobilization of metals associated with different mineral phases. Under these conditions, biochar may have reduced efficacy for reducing heavy metal uptake in wheat.

Short-term biochar manipulation of microbial nitrogen transformation in wheat rhizosphere of a metal contaminated Inceptisol from North China plain.

While metal immobilization had been increasingly reported with biochar soil amendment (BSA), changes in microbial activity and nitrogen (N) transformation in metal contaminated croplands following biochar addition had been insufficiently addressed. In a field experiment, a Pb/Cd contaminated Inceptisol from North China was amended to topsoil with wheat straw biochar at 0 (CK), 20 (C1) and 40 t ha-1 (C2). The changes within two years following BSA were tested in microbial biomass and respiration, and in abundance of N transforming microbial communities and their activities. Corresponding to the results of decreased soil extractable Cd and Pb, significant reductions in qCO2 were found in rhizosphere and bulk soil only under C2 in the first year. The potential nitrification activity was significantly increased by 20-71%, along with an increase in ammonium (by 7-21%) and nitrate (by 21%-70%) concentration, with BSA compared to CK. Meanwhile, N2O production activity was slightly increased (by up to 20%) but N2O reduction activity greatly enhanced (by up to 84%), with a higher ratio of nosZ/(nirS + nirK), under C2 in rhizosphere in both wheat seasons. Whereas, such changes were not remarkable in bulk soil. Moreover, microbial communities were less respondent to biochar in the second year following the addition. Therefore, microbial growth and functioning for N transforming and cycling in metal contaminated soils could be largely improved with BSA at 40 t ha-1. Of course, studies are still deserved to mimic the long term changes with biochar in N cycling of the metal contaminated dry croplands.

The endophytic bacterium Serratia sp. PW7 degrades pyrene in wheat.

This research was conducted to isolate polycyclic aromatic hydrocarbon-degrading (PAH-degrading) endophytic bacteria and investigate their potential in protecting plants against PAH contamination. Pyrene-degrading endophytic bacteria were isolated from plants grown in PAH-contaminated soil. Among these endophytic bacteria, strain PW7 (Serratia sp.) isolated from Plantago asiatica was selected to investigate the suppression of pyrene accumulation in Triticum aestivum L. In the in vitro tests, strain PW7 degraded 51.2% of the pyrene in the media within 14 days. The optimal biodegradation conditions were pH 7.0, 30 °C, and MS medium supplemented with additional glucose, maltose, sucrose, and peptones. In the in vivo tests, strain PW7 successfully colonized the roots and shoots of inoculated (E+) wheat plants, and its colonization decreased pyrene accumulation and pyrene transportation from roots to shoots. Remarkably, the concentration of pyrene in shoots decreased much more than that in roots, suggesting that strain PW7 has the potential for protecting wheat against pyrene contamination and mitigating the threat of pyrene to human health via food consumption.