An Optical Sensing Platform for Beta-Glucosidase Activity Using Protein-Inorganic Hybrid Nanoflowers.

In this work, a convenient and dual-signal readout optical sensing platform for the sensitively and selectively determination of beta-glucosidase (ß-Glu) activity was reported using protein-inorganic hybrid nanoflowers [BSA-Cu3(PO4)2·3H2O] possessing peroxidase-mimicking activity. The nanoflowers (NFs) were facilely synthesized through a self-assembled synthesis strategy at room temperature. The as-prepared NFs could catalytically convert the colorless and non-fluorescent Amplex Red into colored and highly fluorescent resorufin in the presence of hydrogen peroxide via electron transfer process. ß-Glu could hydrolyze cyanogenic glycoside, using amygdalin (Amy) as a model, into cyanide ions (CN-), which can subsequently efficiently suppress the catalytic activity of NFs, accompanied with the fluorescence decrease and the color fading. The concentration of CN- was controlled by ß-Glu-triggered enzymatic reaction of Amy. Thus, a sensing system was established for fluorescent and visual determination of ß-Glu activity. Under the optimum conditions, the present fluorescent and visual bimodal sensing platform exhibited good sensitivity for ß-Glu activity assay with a detection limit of 0.33 U·L-1. The sensing platform was further applied to determinate ß-Glu in real samples and satisfactory results were attained. Additionally, the optical sensing system can potentially be a promising candidate for ß-Glu inhibitors screening.

Soil microbial community responses to short-term nitrogen addition in China's Horqin Sandy Land.

Anthropogenic nitrogen (N) addition has increased soil nutrient availability, thereby affecting ecosystem processes and functions in N-limited ecosystems. Long-term N addition decreases plant biodiversity, but the effects of short-term N addition on soil microbial community is poorly understood. The present study examined the impacts of short-term N addition (NH4NO3) on these factors in a sandy grassland and semi-fixed sandy land in the Horqin Sandy Land. We measured the responses of soil microbial biomass C and N; on soil ß-1,4-glucosidase (BG) and ß-1,4-N-acetylglucosaminidase (NAG) activity; and soil microflora characteristics to N additions gradient with 0 (control), 5 (N5), 10 (N10), and 15 (N15) g N m-2 yr-1. The soil microbial biomass indices, NAG activity, and soil microflora characteristics did not differ significantly among the N levels, and there was no difference at the two sites. The competition for N between plants and soil microbes was not eliminated by short-term N addition due to the low soil nutrient and moisture contents, and the relationships among the original soil microbes did not change. However, N addition increased BG activity in the N5 and N10 additions in the sandy grassland, and in the N5, N10, and N15 additions in the semi-fixed sandy land. This may be due to increased accumulation and fixation of plant litter into soils in response to N addition, leading to increased microbial demand for a C source and increased soil BG activity. Future research should explore the relationships between soil microbial community and N addition at the two sites.

Impact of Ag2S NPs on soil bacterial community - A terrestrial mesocosm approach.

Soils might be a final sink for Ag2S nanoparticles (NPs). Still, there are limited data on their effects on soil bacterial communities (SBC). To bridge this gap, we investigated the effects of Ag2S NPs (10 mg kg-1 soil) on the structure and function of SBC in a terrestrial indoor mesocosm, using a multi-species design. During 28 days of exposure, the SBC function-related parameters were analysed in terms of enzymatic activity, community level physiological profile, culture of functional bacterial groups [phosphorous-solubilizing bacteria (P-SB) and heterotrophic bacteria (HB)], and SBC structure was analysed by 16S rRNA gene-targeted denaturing gradient gel electrophoresis. The SBC exposed to Ag2S NPs showed a significative decrease of functional parameters, such as ß-glucosidase activity and L-arginine consumption, and increase of the acid phosphatase activity. At the structural level, significantly lower richness and diversity were detected, but at later exposure times compared to the AgNO3 treatment, likely because of a low dissolution rate of Ag2S NPs. In fact, stronger effects were observed in soils spiked with AgNO3, in both functional and structural parameters. Changes in SBC structure seem to negatively correlate with parameters related to phosphorous (acid phosphatase activity) and carbon cycling (abundance of HB, P-SB, and ß-glucosidase activity). Our results indicate a significant effect of Ag2S NPs on SBC, specifically on parameters related to carbon and phosphorous cycling, at doses as low as 10 mg kg-1 soil. These effects were only observed after 28 days, highlighting the importance of long-term exposure experiments for slowly dissolving NPs.

Efficacy of fluopyram applied by chemigation on controlling eggplant root-knot nematodes (Meloidogyne spp.) and its effects on soil properties.

The root-knot nematode (Meloidogyne spp.) is one of the major challenges in eggplant (Solanum melongena L.) production. Fluopyram, known to be an effective fungicide, is also used for controlling root-knot nematode. However, in China, little information is currently available regarding the efficacy of fluopyram via chemigation against root-knot nematode and its effects on soil properties. For this, the objective of this work was to test mortality of root-knot nematode, functional diversity of soil microbial community, activity of soil enzyme after fluopyram applicated by chemigation. The results of two field experiments revealed that concentration of 60 g·ha-1 fluopyram applied with 200 L·ha-1 irrigation water at 2 L·h-1 flow velocity was the most effective chemigation parameters for controlling eggplant against root-knot nematode. The functional diversity of the soil microbial community was significantly affected by fluopyram. The activities of soil urease and ß-glucosidase decreased during the initial stages but recovered at later stages. In brief, fluopyram has advantageous for the efficient control of root-knot nematode with no deleterious effects on soil properties as well as chemigation is positive for application in karst landscape in Guangxi.

1H, 13C, 15N backbone and IVL methyl group resonance assignment of the fungal ß-glucosidase from Trichoderma reesei.

ß-glucosidases have received considerable attention due to their essential role in bioethanol production from lignocellulosic biomass. ß-glucosidase can hydrolyse cellobiose in cellulose degradation and its low activity has been considered as one of the main limiting steps in the process. Large-scale conversions of cellulose therefore require high enzyme concentration which increases the cost. ß-glucosidases with improved activity and thermostability are therefore of great commercial interest. The fungus Trichoderma reseei expresses thermostable cellulolytic enzymes which have been widely studied as attractive targets for industrial applications. Genetically modified ß-glucosidases from Trichoderma reseei have been recently commercialised. We have developed an approach in which screening of low molecular weight molecules (fragments) identifies compounds that increase enzyme activity and are currently characterizing fragment-based activators of TrBgl2. A structural analysis of the 55 kDa apo form of TrBgl2 revealed a classical (α/ß)8-TIM barrel fold. In the present study we present a partial assignment of backbone chemical shifts, along with those of the Ile (I)-Val (V)-Leu (L) methyl groups of TrBgl2. These data will be used to characterize the interaction of TrBgl2 with the small molecule activators.

Enzyme-triggered fluorescence turn-off/turn-on of carbon dots for monitoring ß-glucosidase and its inhibitor in living cells.

Energy transfer engineering based on fluorescent probes for directly sensing enzyme activities are in great demand as enzyme-mediated transformations, which are central to all biological processes. Here, a fluorescence carbon dot (CD)-based assay exhibiting selective responses to the quantitation of ß-glucosidase and the effect of its inhibitor was developed. The most common substrate, para-nitrophenyl-ß-d-glucopyranoside (pNPG) was hydrolyzed by ß-glucosidase to release p-nitrophenol (pNP), which can efficiently quench fluorescence of CDs via an inner filter effect and electron transfer. However, in the presence of inhibitors of ß-glucosidase, the fluorescence intensity gradually recovered as the concentration of inhibitors increased. Therefore, the enzyme-triggered fluorescence turn-off/turn-on of specific CDs successfully achieved sensitive detection of ß-glucosidase and monitored the effect of its inhibitors. This new strategy was applied to detect ß-glucosidase and monitor ß-glucosidase inhibitor in hepatoma cells using cell imaging. All results suggest that the new method is sensitive and promising for use in cancer diagnosis and treatment.

A "turn off-on" fluorescent nanoprobe consisting of CuInS2 quantum dots for determination of the activity of ß-glucosidase and for inhibitor screening.

A fluorescent "turn off-on" nanoprobe is described for highly sensitive and selective determination of the activity of the enzyme ß-glucosidase (ß-Glu). Firstly, cysteine modified CuInS2 quantum dots (Cys-CuInS2 QDs) were prepared from indium(III) and copper(II) salts and the presence of thiourea. The red fluorescence of the Cys-CuInS2 QDs, with excitation/emission maxima at 590/656 nm, is quenched by Cu(II). However, in the presence of ß-Glu and the cyanogenic glycoside, enzymatic hydrolysis leads to the formation of cyanide. The latter competitively binds to Cu(II) owing to its high affinity for cyanide. This restores the fluorescence of the Cys-CuInS2 QDs. Under the optimum conditions, fluorescence increases linearly in the 0.5-700 U·L-1 ß-Glu activity range. The detection limit is 0.2 U·L-1. The nanoprobe was applied to analyze spiked soil samples, and satisfactory results were obtained. The average recoveries of ß-Glu were in the range of 96-103%, and the RSD was lower than 4.0%. The fluorescent probe can also be used to screen for ß-Glu inhibitors as demonstrated for castanospermine as an example. Graphical abstractSchematic representation of the fluorescent nanoprobe for ß-glucosidase activity detection and inhibitor screening by taking advantage of the fluorescence (FL) "turn-off" and "turn-on" feature of cysteine capped CuInS2 quantum dots (Cys-CuInS2 QDs).

Application of wood biochar in polluted soils stabilized the toxic metals and enhanced wheat (Triticum aestivum) growth and soil enzymatic activity.

Biochar is a stable carbonaceous by-product of pyrolysis and can be used for toxic metals (TMs) retention in polluted soil. Wheat (Triticum aestivum) was grown in three polluted soils collected from Chenzhou (CZ), Tongguan (TG) and Fengxian (FX), China. Wood biochar (WBC) was applied at 0, 0.5, 1.0 and 2.0% to each pot filled with 2 kg polluted soil. The results showed that WBC was efficient to alter soil pH and electrical conductivity (EC). The changes in soil pH and EC had a direct relationship with the immobilization and phytostabilization of TMs in the three soils. The bioavailable TMs (Zn, Pb, Cd, and Cu) were reduced in the soil after WBC amendments due to ion exchange, precipitates of metal-carbonates and metal-phosphates, and chemisorption on WBC surface. The reduction in the bioavailable TMs content also resulted in the diminution in TMs shoot uptake in wheat. Similarly, the TMs uptake in wheat root were also reduced as a result of WBC application. The reduction in bioavailable TMs and the release of essential nutrients and base cations from the WBC also increased the wheat shoot and root dry biomasses production. The application of WBC in polluted soil also improved soil health and the urease and ß-glucosidase enzymes were also enhanced. The results concluded that WBC was efficient to reduce the bioavailability of TMs and shoot and root uptake, improved wheat dry biomasses production and soil enzymatic activities in industrial and smelter/mines polluted soils.

Effect of xylitol on salivary ß-glucosidase in humans.

Dental biofilm - in which a diverse set of microorganisms are embedded in a complex polysaccharide matrix that adheres to oral components - is one of the most complex microbial communities in the human body. As biofilm formation is related to oral infections, such as caries and periodontal diseases, strategies for biofilm control are crucial for maintaining oral health. Xylitol, a synthetic sugar used as a sucrose substitute, has been shown to reduce biofilm formation. However, its precise mechanism of action on biofilm reduction has so far not been elucidated. Previous studies demonstrate that bacterial ß-glucosidase action is crucial for biofilm formation. Here, we investigated the correlation between salivary ß-glucosidase activity and dental plaque occurrence. We found a positive correlation between enzymatic activity and the presence of dental biofilm. We observed that xylitol inhibits ß-glucosidase in human saliva. Kinetic studies also confirmed that xylitol acts as a mixed type inhibitor of salivary ß-glucosidase. Based on our data, we suggest that xylitol impairs oral biofilm formation by the inhibition of bacterial ß-glucosidase, which is essential for biofilm formation in the oral cavity.

A simplified and miniaturized glucometer-based assay for the detection of ß-glucosidase activity.

ß-Glucosidase activity assays constitute an important indicator for the early diagnosis of neonatal necrotizing enterocolitis and qualitative changes in medicinal plants. The drawbacks of the existing methods are high consumption of both time and reagents, complexity in operation, and requirement of expensive instruments and highly trained personnel. The present study provides a simplified, highly selective, and miniaturized glucometer-based strategy for the detection of ß-glucosidase activity. Single-factor experiments showed that optimum ß-glucosidase activity was exhibited at 50 °C and pH 5.0 in a citric acid-sodium citrate buffer when reacting with 0.03 g/mL salicin for 30 min. The procedure for detection was simplified without the need of a chromogenic reaction. Validation of the analytical method demonstrated that the accuracy, precision, repeatability, stability, and durability were good. The linear ranges of ß-glucosidase in a buffer solution and rat serum were 0.0873-1.5498 U/mL and 0.4076-2.9019 U/mL, respectively. The proposed method was free from interference from ß-dextranase, snailase, ß-galactosidase, hemicellulase, and glucuronic acid released by baicalin. This demonstrated that the proposed assay had a higher selectivity than the conventional dinitrosalicylic acid (DNS) assay because of the specificity for salicin and unique recognition of glucose by a personal glucose meter. Miniaturization of the method resulted in a microassay for ß-glucosidase activity. The easy-to-operate method was successfully used to detect a series of ß-glucosidases extracted from bitter almonds and cultured by Aspergillus niger. In addition, the simplified and miniaturized glucometer-based assay has potential application in the point-of-care testing of ß-glucosidase in many fields, including medical diagnostics, food safety, and environmental monitoring.

Teasing apart the different size pools of extracellular enzymatic activity in the ocean.

Extracellular enzymatic activity (EEA) is performed by cell-associated and cell-free (i.e., "dissolved") enzymes. This cell-free fraction is operationally defined as passing through a 0.22 µm filter. The contribution of cell-free to total EEA is comparable to the cell-associated counterpart, so it is critical to understand what controls the relative importance of cell-free versus cell-associated EEA. However, attempts to tease apart the contribution of EEAs in the so-called dissolved fraction (<0.22 µm) in general, and of the nanoparticle size fraction (0.020-0.20 µm) in particular, to the total EEA pool are lacking. Here we performed experiments with Northern and Southern Hemisphere coastal waters to characterize the potential contribution of that nanoparticle fraction to the total EEA fraction of alkaline phosphatase, beta-glucosidase and leucine aminopeptidase. We found a significant contribution (in both hemispheres) of the nanoparticle fraction to the total EEA pool (up to 53%) that differed depending on the enzyme type and location. Collectively, our results indicate that a significant fraction of the so-called "dissolved EEA" is not really dissolved but associated to nanoparticles, colloidal nanogels and/or viruses. Thus, the total marine EEA pool can actually be divided into a cell-associated, undissolved-cell-free (associated to nano-particle of different origins such as viruses and nanogels) and a dissolved-cell-free pools. Our results also imply that the dissolved EEA pool is more complex than thus far anticipated. Future research will be now needed to further characterize the factors controlling the relative importance of these different pools of EEA, which are key in the recycling of organic matter in the ocean.

Varying Inocula Permutations (Aspergillus oryzae and Bacillus amyloliquefaciens) affect Enzyme Activities and Metabolite Levels in Koji.

In this study, we investigated the altered enzymatic activities and metabolite profiles of koji fermented using varying permutations of Aspergillus oryzae and/or Bacillus amyloliquefaciens. Notably, the protease and ß-glucosidase activities were manifold increased in co-inoculated (CO) koji samples (co-inoculation of A. oryzae and B. amyloliquefaciens). Furthermore, gas chromatography-mass spectrometry (GC-MS)-based metabolite profiling indicates that levels of amino acids, organic acids, sugars, sugar alcohols, fatty acids, nucleosides, and vitamins were distinctly higher in CO, SA (sequential inoculation of A. oryzae, followed by B. amyloliquefaciens), and SB (sequential inoculation of B. amyloliquefaciens, followed by A. oryzae). The multivariate principal component analysis (PCA) plot based on GC-MS datasets indicated a clustered pattern for MA and MB (koji samples inoculated either with A. oryzae or B. amyloliquefaciens) across PC2 (20.0%). In contrast, the CO, SA, and SB metabolite profiles displayed segregated patterns across PLS1 (22.2%) and PLS2 (21.1%) in the partial least square discriminant analysis (PLS-DA) model. Intriguingly, the observed disparity in the levels of primary metabolites was engendered largely by higher relative levels of sugars and sugar alcohols in MA, SA, and CO koji samples, which was commensurate with the relative amylase activities in respective samples. Collectively, the present study emphasizes the utility of integrated biochemical and metabolomic approaches for achieving the optimal permutation of fermentative inocula for industrial koji preparation.

Effects of the herbicide mesotrione on soil enzyme activity and microbial communities.

Mesotrione (2-[4-(methylsulfonyl)-2-nithobenzoyl]-1, 3-cyclohexanedione) is a selective triketone herbicide that has been widely used in corn production for the past 15 years. However, its potential for risk to soil ecosystems is poorly documented. The present study investigated the soil enzyme activity and soil microbial community responses to a 20 days' mesotrione exposure at doses of 0.1, 1.0 and 5.0 mg/kg. On days 2, 5, 10 and 20, activities of soil ß-glucosidase, urease and acid phosphatase, soil microbe abundances, soil microbial community structure and abundance of the AOA-amoA and AOB-amoA genes were measured. Results showed that activities of urease and acid phosphatase were relatively stable, with no difference found between the mesotrione-treated group and control at the end of exposure. But ß-glucosidase activity was reduced in the 5.0 mg/kg mesotrione treatment. In the 1.0 and 5.0 mg/kg mesotrione-treated soil, abundance of bacteria, fungi and actinomycetes all were reduced. In the 0.1 mg/kg mesotrione-treated soil, only fungi abundance was reduced by the end of the exposure. The analysis of terminal restriction fragment length polymorphism (T-RFLP) revealed soil microbial community structure could be affected by mesotrione at all experimental doses, and microbial diversity declined slightly after mesotrione exposure. Abundance of AOA-amoA and AOB-amoA genes were reduced markedly in 1.0 and 5.0 mg/kg mesotrione-treated soil. The present study suggests that mesotrione at higher doses might induce negative impacts on soil microbes, a finding which merits additional research and which should be accounted for when application of this herbicide is considered.

Coumarin-based, switchable fluorescent substrates for enzymatic bacterial detection.

Enzymatically-switchable fluorescent substrates, such as the commercially available 4-methyl umbelliferones (4-MU) are used as standard indicators of enzymatic activity for the detection of various microorganisms and pathogens. However, a major disadvantage of 4-MU is its relatively high pKa leading to only partial dissociation of the fluorescent anion under the conditions where the enzymes are most effective (pH 6-6.5). Here we present a method for new, enzymatically-switchable, fluorescent substrates with improved photo-physico/chemical properties. The lead derivative, 4-AAU, shows excellent solubility in aqueous media (0.81 mg/mL) when compared to 4-MU (0.16 mg/mL), significantly improved quantum yield and wider dynamic range of its fluorescence properties. The corresponding bacterial substrate ß-4-AAUG showed superior selectivity in the detection of clinically relevant amounts of E. coli, Enterococcus and K. pneumonia (1 CFU). The fluorescence intensity of ß-4-AAUG was almost 5 times higher than that of the standard, the detection was possible in reasonably short time (∼ 2.5 h) and with excellent sensitivity.

[Effects of Long-term Fertilization on Enzyme Activities in Profile of Paddy Soil Profiles].

The enzyme activity, which is closely related to soil material cycling (mineralization, transformation, etc.), can reflect soil quality and nutrient status. In order to explore the effect of long-term fertilization on the enzyme activity in paddy soil profile (0-40 cm), soils with organic fertilizer and inorganic fertilizer, and non-fertilized soils were selected, and the carbon and nitrogen contents, and the activities of ß-1,4-glucosidase (BG), and ß-1,4-N-acetylglucosaminidase (NAG) in 10cm depths of soil were analyzed. The results showed that the activities of BG and NAG in the soils treated with inorganic fertilizer and organic fertilizer increased by 0.73-47.87 nmol·(g·h)-1 and 1.33-128.81 nmol·(g·h)-1, and 0.19-9.72 nmol·(g·h)-1 and 0.92-57.66 nmol·(g·h)-1, respectively, compared to those for non-fertilized soil. Soil enzyme activity decreased with increasing soil depth. Soil enzyme activity in soil from 0-20 cm was significantly higher than that of soil from 20-40 cm. Soil enzyme activities were significantly affected by long term fertilization at different soil depths. RDA analysis showed that soil carbon and nitrogen contents had significant positive relationships with the activities of BG and NAG in the 0-20 cm soil profiles, however, negative relationships were observed in the 20-40 cm soil profiles. The long-term application of organic fertilizer significantly increased soil biomass and enzyme activity, both of which decreased with the increase in soil depth. Long-term fertilization could increase soil nutrient contents, microbial biomass, and extracellular enzyme activities, which has important theoretical significance for optimizing farmland fertilizer management and improving soil productivity.

Impacts of Phragmites australis Invasion on Soil Enzyme Activities and Microbial Abundance of Tidal Marshes.

The rapid expansion of Phragmites australis in brackish marshes of the East Coast of the USA has drawn much attention, because it may change vegetation diversity and ecosystem functions. In particular, higher primary production of Phragmites than that of other native species such as Spartina patens and Schoenoplectus americanus has been noted, suggesting possible changes in carbon storage potential in salt marshes. To better understand the long-term effect of the invasion of Phragmites on carbon storage, however, information on decomposition rates of soil organic matter is essential. To address this issue, we compared microbial enzyme activities and microbial functional gene abundances (fungi, laccase, denitrifier, and methanogens) in three depths of soils with three different plants in a brackish marsh in Maryland, USA. Laccase and phenol oxidase activities were measured to assess the decomposition potential of recalcitrant carbon while ß-glucosidase activity was determined as proxy for cellulose decomposition rate. Microbial activities near the surface (0-15 cm) were the highest in Spartina-community sites followed by Phragmites- and Schoenoplectus-community sites. A comparison of stable isotopic signatures (δ13C and δ15N) of soils and plant leaves suggests that deep organic carbon in the soils mainly originated from Spartina, and only the surface soils may have been influenced by Phragmites litter. In contrast, fungal, laccase, and denitrifier abundances determined by real-time qPCR exhibited no discernible patterns among the surface soils of the three vegetation types. However, the abundance of methanogens was higher in the deep Phragmites-community soil. Therefore, Phragmites invasion will accelerate CH4 emission by greater CH4 production in deep soils with abundant methanogens, although enzymatic mechanisms revealed the potential for larger C accumulation by Phragmites invasion in salt marshes in the east coast of the USA.

Regular enzyme recovery enhances cellulase production by Trichoderma reesei in fed-batch culture.

OBJECTIVE: To protect the enzymes during fed-batch cellulase production by means of partial enzyme recovery at regular intervals. RESULTS: Extracellular enzymes were partially recovered at the intervals of 1, 2, or 3 days. Mycelia were also removed to avoid contamination. Increases in the total harvested cellulase (24-62%) and ß-glucosidase (22-76%) were achieved. In fermentor cultivation when the enzymes were recovered every day with 15% culture broth. The total harvested cellulase and ß-glucosidase activity increased by 43 and 58%, respectively, with fungal cell concentration maintained at 3.5-4.5 g l-1. CONCLUSION: Enzyme recovery at regular intervals during fed-batch cellulase cultivation could protect the enzyme in the culture broth and enhance the enzyme production when the fungal cell concentration is maintained in a reasonable range.

Functional Characterization of CsBGlu12, a ß-Glucosidase from Crocus sativus, Provides Insights into Its Role in Abiotic Stress through Accumulation of Antioxidant Flavonols.

Glycosylation and deglycosylation are impressive mechanisms that allow plants to regulate the biological activity of an array of secondary metabolites. Although glycosylation improves solubility and renders the metabolites suitable for transport and sequestration, deglycosylation activates them to carry out biological functions. Herein, we report the functional characterization of CsBGlu12, a ß-glucosidase from Crocus sativus. CsBGlu12 has a characteristic glucoside hydrolase 1 family (α/ß)8 triose-phosphate isomerase (TIM) barrel structure with a highly conserved active site. In vitro enzyme activity revealed that CsBGlu12 catalyzes the hydrolysis of flavonol ß-glucosides and cello-oligosaccharides. Site-directed mutagenesis of any of the two conserved catalytic glutamic acid residues (Glu200 and Glu414) of the active site completely abolishes the ß-glucosidase activity. Transcript analysis revealed that Csbglu12 is highly induced in response to UV-B, dehydration, NaCl, methyl jasmonate, and abscisic acid treatments indicating its possible role in plant stress response. Transient overexpression of CsBGlu12 leads to the accumulation of antioxidant flavonols in Nicotiana benthamiana and confers tolerance to abiotic stresses. Antioxidant assays indicated that accumulation of flavonols alleviated the accretion of reactive oxygen species during abiotic stress conditions. ß-Glucosidases are known to play a role in abiotic stresses, particularly dehydration through abscisic acid; however, their role through accumulation of reactive oxygen species (ROS) scavenging flavonols has not been established. Furthermore, only one ß-glucosidase 12 homolog has been characterized so far. Therefore, this work presents an important report on characterization of CsBGlu12 and its role in abiotic stress through ROS scavenging.

A comparative study on the influence of different organic amendments on trace element mobility and microbial functionality of a polluted mine soil.

A mine soil heavily polluted with zinc and cadmium was employed to evaluate the capacity of organic amendments of different origin to simultaneously reduce soil trace element mobility and enhance soil microbial functionality. With this aim, four organic products, namely olive processing solid waste (OPSW), municipal solid waste compost (MSWC), leonardite and peat, were applied individually at different doses (0, 1, 2 and 5%) to mine soil under controlled laboratory conditions. Extraction studies and analysis of soil microbiological parameters (basal soil respiration and dehydrogenase, ß-glucosidase, urease, arylsulfatase and acid and alkaline phosphatase activities) were performed to assess the effect of such amendments on soil restoration. Their ability to decrease mine soil mobile trace element contents followed the sequence MSWC > OPSW > peat > leonardite, with the former achieving reduction levels of 78 and 73% for Zn and Cd, respectively, when applied at a dose of 5%. This amendment also showed a good performance to restore soil microbial functionality. Thus, basal soil respiration and dehydrogenase, urease and alkaline phosphatase activities experienced increases of 187, 79, 42 and 26%, respectively, when mine soil was treated with 5% MSWC. Among tested organic products, MSWC proved to be the best amendment to perform both the chemical and the microbial soil remediation.

Exoenzyme activity in contaminated soils before and after soil washing: ß-glucosidase activity as a biological indicator of soil health.

It is essential to remediate or amend soils contaminated with various heavy metals or pollutants so that the soils may be used again safely. Verifying that the remediated or amended soils meet soil quality standards is an important part of the process. We estimated the activity levels of eight soil exoenzymes (acid phosphatase, arylsulfatase, catalase, dehydrogenase, fluorescein diacetate hydrolase, protease, urease, and ß-glucosidase) in contaminated and remediated soils from two sites near a non-ferrous metal smelter, using colorimetric and titrimetric determination methods. Our results provided the levels of activity of soil exoenzymes that indicate soil health. Most enzymes showed lower activity levels in remediated soils than in contaminated soils, with the exception of protease and urease, which showed higher activity after remediation in some soils, perhaps due to the limited nutrients available in remediated soils. Soil exoenzymes showed significantly higher activity in soils from one of the sites than from the other, due to improper conditions at the second site, including high pH, poor nutrient levels, and a high proportion of sand in the latter soil. Principal component analysis revealed that ß-glucosidase was the best indicator of soil ecosystem health, among the enzymes evaluated. We recommend using ß-glucosidase enzyme activity as a prior indicator in estimating soil ecosystem health.