Magnesium-Isotope Fractionation in Chlorophyll-a Extracted from Two Plants with Different Pathways of Carbon Fixation (C3, C4).

Relatively few studies have been focused so far on magnesium-isotope fractionation during plant growth, element uptake from soil, root-to-leaves transport and during chlorophylls biosynthesis. In this work, maize and garden cress were hydroponically grown in identical conditions in order to examine if the carbon fixation pathway (C4, C3, respectively) might have impact on Mg-isotope fractionation in chlorophyll-a. The pigment was purified from plants extracts by preparative reversed phase chromatography, and its identity was confirmed by high-resolution mass spectrometry. The green parts of plants and chlorophyll-a fractions were acid-digested and submitted to ion chromatography coupled through desolvation system to multiple collector inductively coupled plasma-mass spectrometry. Clear preference for heavy Mg-isotopes was found in maize green parts (∆26Mgplant-nutrient 0.65, 0.74 for two biological replicates, respectively) and in chlorophyll-a (∆26Mgchlorophyll-plant 1.51, 2.19). In garden cress, heavy isotopes were depleted in green parts (∆26Mgplant-nutrient (-0.87)-(-0.92)) and the preference for heavy isotopes in chlorophyll-a was less marked relative to maize (∆26Mgchlorophyll-plant 0.55-0.52). The observed effect might be ascribed to overall higher production of energy in form of adenosine triphosphate (ATP), required for carbon fixation in C4 compared to C3, which could reduce kinetic barrier and make equilibrium fractionation prevailing during magnesium incorporation to protoporphyrin ring.

Fungal biodegradation and multi-level toxicity assessment of vinasse from distillation of winemaking by-products.

The wastewaters from distilleries of winemaking by-products, a scarcely studied type of vinasse, were treated by white-rot fungal strains from species Irpex lacteus, Ganoderma resinaceum, Trametes versicolor, Phlebia rufa and Bjerkandera adusta. The main objectives of this study were to evaluate fungal performance during vinasse biodegradation, their enzyme patterns and ecotoxicity evolution throughout treatment. Despite all strains were able to promote strong (>80%) dephenolization and reduction of total organic carbon (TOC), P. rufa was less affected by vinasse toxicity and exhibit better decolorization. In batch cultures at 28 °C and pH 4.0, the first phase of P. rufa biodegradation kinetics was characterized by strong metabolic activity with simultaneous depletion of TOC, phenolics and sugars. The main events of second phase are the increase of peroxidases production after the peak of laccase activity, and strong color removal. At the end of treatment, it was observed highly significant (p < 0.001) abatement of pollution parameters (83-100% removal). Since water reclamation and reuse for e.g. crop irrigation is a priority issue, vinasse ecotoxicity was assessed with bioindicators representing three different phylogenetic and trophic levels: a marine bacterium (Aliivibrio fischeri), a freshwater microcrustacean (Daphnia magna) and a dicotyledonous macrophyte (Lepidium sativum). It was observed significant (p < 0.05) reduction of initial vinasse toxicity, as evaluated by these bioindicators, deserving special mention an almost complete phytotoxicity elimination.

Monochromatic lights-induced trends in antioxidant and antidiabetic polyphenol accumulation in in vitro callus cultures of Lepidium sativum L.

Lepidium sativum L. is an important edible, herbaceous plant with huge medicinal value as cardio-protective, hepatoprotective and antitumor agent. This study was designed and performed to investigate biosynthesis of plant's active ingredients in callus cultures of L. sativum in response to the exposure of multi spectral lights. Optimum biomass accumulation (15.36 g/L DW), total phenolic and flavonoid contents (TPC; 47.43 mg/g; TFC; 9.41 mg/g) were recorded in calli placed under white light (24 h) compared to rest of the treatments. Antioxidant enzymatic activities i.e. superoxide dismutase and peroxidase were found optimum in cultures exposed to green light (SOD; 0.054 nM/min/mg FW, POD; 0.501 nM/min/mg FW). Phytochemical analysis further confirmed the potential influence of white light exposure on enhanced production of plant's metabolites. Significant enhancement level of major metabolic compounds such as chlorogenic acid (7.20 mg/g DW), quercetin (22.08 mg/g DW), kaempferol (7.77 mg/g DW) and minor compounds including ferulic acid, sinapic acid, protocatechuic acid, vanillic acid and caffeic acid were recorded in white light compared to control (photoperiod), whereas blue light increased the p-coumaric acid accumulation. Moreover, callus cultures of this plant under white light (24 h) showed highest in vitro based anti-diabetic and antioxidant activities compared to other conditions. Finding of our current study revealed that multi spectral lights are proved to be an effective strategy for enhancing metabolic quantity of antioxidant and anti-diabetic bioactive compounds in callus cultures of L. sativum L.

Plant Growth Biostimulants Based on Different Methods of Seaweed Extraction with Water.

We explored two methods for obtaining aqueous extracts: boiling and soaking of Baltic seaweeds (EB and ES, resp.). Algal extracts were characterized in terms of polyphenols, micro- and macroelements, lipids content, and antibacterial properties. The utilitarian properties were examined in the germination tests on Lepidium sativum for three extract dilutions (0.5, 2.5, and 10%). It was found that the extracts were similar in micro- and macroelement concentrations. Water was proved to be a good solvent to extract phenolic compounds. The algal extract produced by soaking biomass did not show inhibitory effect on Escherichia coli and Staphylococcus aureus. Only the boiled extract had an inhibitory activity against E. coli. Germination tests revealed a positive influence of the bioproducts on the cultivated plants. In the group treated with 10% EB, plants were 13% longer than in the control group; the content of elements B, Mo, Zn, and Na in the group treated with 10% ES was higher by 76%, 48%, 31%, and 59% than in the control group, respectively; the content of chlorophyll was 2.5 times higher in 0.5% ES than in the control group. Extracts showed the slight impact on the morphology of plants.

Phytotoxicity of ionic, micro- and nano-sized iron in three plant species.

Potential environmental impacts of engineered nanoparticles (ENPs) can be understood taking into consideration phytotoxicity. We reported on the effects of ionic (FeCl3), micro- and nano-sized zerovalent iron (nZVI) about the development of three macrophytes: Lepidium sativum, Sinapis alba and Sorghum saccharatum. Four toxicity indicators (seed germination, seedling elongation, germination index and biomass) were assessed following exposure to each iron concentration interval: 1.29-1570mg/L (FeCl3), 1.71-10.78mg/L (micro-sized iron) and 4.81-33,560mg/L (nano-iron). Exposure effects were also observed by optical and transmission electron microscopy. Results showed that no significant phytotoxicity effects could be detected for both micro- and nano-sized zerovalent irons, including field nanoremediation concentrations. Biostimulation effects such as an increased seedling length and biomass production were detected at the highest exposure concentrations. Ionic iron showed slight toxicity effects only at 1570mg/L and, therefore, no median effect concentrations were determined. By microscopy, ENPs were not found in palisade cells or xylem. Apparently, aggregates of nZVI were found inside S. alba and S. saccharatum, although false positives during sample preparation cannot be excluded. Macroscopically, black spots and coatings were detected on roots of all species especially at the most concentrated treatments.

Diversity of organotrophic bacteria, activity of dehydrogenases and urease as well as seed germination and root growth Lepidium sativum, Sorghum saccharatum and Sinapis alba under the influence of polycyclic aromatic hydrocarbons.

Polycyclic aromatic hydrocarbons are organic compounds with highly toxic, carcinogenic, and mutagenic properties, which adversely affect the basic biological parameters of the soil, including the count of microorganisms, and the enzymatic activity. In addition to disturbances to the biological activity of the soil, PAHs may also exhibit toxic effects on plants. In view of the above, the study involved testing aimed at the determination of the effects of polycyclic aromatic hydrocarbons in a form of naphthalene, phenanthrene, anthracene and pyrene on the count, colony development (CD) index, ecophysiological (EP) diversity index of organotrophic bacteria, and the activity of soil dehydrogenases and soil urease. Moreover, an attempt was made to determine the soil's resistance based on the activity of the above-listed enzymes, and the effect of polycyclic aromatic hydrocarbons on seed germination and root growth was assessed by Lepidium sativum, Sorghum saccharatum, and Sinapis alba. In addition, the species of bacteria found in a soil subjected to strong pressure of polycyclic aromatic hydrocarbons were isolated. The experiment was performed in a laboratory on samples of loamy sand. Polycyclic aromatic hydrocarbons were introduced into the soil in an amount of 0, 1000, 2000, and 4000 mg kg(-1) of soil dry matter. Germination and growth of cress (L. sativum), white mustard (S. alba), and sweet sorghum (S. saccharatum) were determined using Phytotoxkit tests. It was found that the tested PAHs increased the average colony counts of organotrophic soil bacteria; pyrene did so to the greatest extent (2.2-fold relative to non-contaminated soil), phenanthrene to the smallest extent (1.4-fold relative to non-contaminated soil). None of the PAHs changed the value of the bacterial colony development (CD) index, while anthracene and pyrene increased the value of the eco-physiological (EP) diversity indicator. PAHs lowered the activity of the tested enzymes. The activity of dehydrogenases was dependent on a greater extent by the type of hydrocarbon (54.56%) rather than by the dose (10.64%), while for the activity of urease, it was the opposite. The greater extent was dependent on dose (95.42%) rather than by type (0.21%). Dehydrogenases are characterised by greater resistance to the action of PAHs than urease. Based on seed germination and root growth, it has shown that S. alba is best suited, being the most vulnerable plant, while S. saccharatum is the least suited. Subjecting a soil to strong pressure of PAHs leads to disturbances to the biological parameters of the soil, seed germination, and root growth L. sativum, S. saccharatum, and S. alba.

Cadmium at high dose perturbs growth, photosynthesis and nitrogen metabolism while at low dose it up regulates sulfur assimilation and antioxidant machinery in garden cress (Lepidium sativum L.).

Metal contamination of soils has become a worldwide problem and great environmental threat, as these metals accumulate in soils and plants in excess, and enter the food chain. Increased cadmium (Cd) uptake from contaminated soils leads to altered plant metabolism and limits the crop productivity. The experimental crop, Lepidium sativum L. (Garden Cress, Family: Brassicaceae) is a medicinally and economically important plant. An experiment was conducted to examine the effect of different concentrations of Cd (0, 25, 50 or 100 mg kg(-1) soil) on the performance of L. sativum. Cd accumulation in roots and leaves (roots>leaves) increased with the increaseing Cd concentration in soil. High Cd concentration (100mg Cd kg(-1) soil) inhibited the leaf area and plant dry mass and significant decline in net photosynthetic rate (P(N)), stomatal conductance (gs), intercellular CO(2) (Ci), chlorophyll (Chl a, Chl b, total Chl) content, carbonic anhydrase (CA; E.C. 4.2.1.1) activity, nitrate reductase (NR; E.C. 1.6.6.1) activity and nitrogen (N) content was also observed. However, ATP-sulfurylase (ATP-S; EC. 2.7.7.4) activity, sulfur (S) content and activities of antioxidant enzymes such as superoxide dismutase (SOD; E.C. 1.15.1.1); catalase (CAT; E.C. 1.11.1.6); ascorbate peroxidase (APX; E.C. 1.11.1.11) and glutathione reductase (GR; E.C. 1.6.4.2) and glutathione (GSH) content were increased. Specifically, the decrease in NR activity and N content showed that Cd affects N metabolism negatively; whereas, the increase in ATP-S activity and S content suggests the up-regulation of S assimilation pathway for possible Cd tolerance in coordination with enhanced activities of antioxidant enzymes and GSH. High Cd concentration (100mg Cd kg(-1) soil) perturbs the L. sativum growth by interfering with the photosynthetic machinery and disrupting the coordination between carbon, N and S metabolism. On the other hand, at low Cd concentration (25mg Cd kg(-1) soil) co-ordination of S and N metabolism complemented to the antioxidant machinery to protect the growth and photosynthesis of L. sativum plants.

Carbon mineralization and distribution of nutrients within different particle-size fractions of commercially produced olive mill pomace.

Composting is a realistic option for disposal of olive mill pomace (OMP) by making it suitable as a soil amendment for organic farming. The chemical and physical characteristics and contribution of particle-size fractions to total nutrients and carbon mineralization of seven commercial composts of OMP (COMP) were investigated. Higher proportions of manure, co-composted with OMP, reduced the organic matter (OM), total carbon and C:N ratio of the product, but increased the content of nutrients and fine particles. The fine particles had higher nutrient contents, but less OM and carbon and, unlike larger particles, did not exhibit any phytotoxicity. Less than 1.5% of added carbon was mineralized in whole compost, but a lower rate was found with larger particles. Separation of COMP by particle size fractionation and application as a soil conditioner is recommended for better optimization of COMP with the <1mm fraction providing the higher quality compost.

Regulation of seed germination in the close Arabidopsis relative Lepidium sativum: a global tissue-specific transcript analysis.

The completion of germination in Lepidium sativum and other endospermic seeds (e.g. Arabidopsis [Arabidopsis thaliana]) is regulated by two opposing forces, the growth potential of the radicle (RAD) and the resistance to this growth from the micropylar endosperm cap (CAP) surrounding it. We show by puncture force measurement that the CAP progressively weakens during germination, and we have conducted a time-course transcript analysis of RAD and CAP tissues throughout this process. We have also used specific inhibitors to investigate the importance of transcription, translation, and posttranslation levels of regulation of endosperm weakening in isolated CAPs. Although the impact of inhibiting translation is greater, both transcription and translation are required for the completion of endosperm weakening in the whole seed population. The majority of genes expressed during this process occur in both tissues, but where they are uniquely expressed, or significantly differentially expressed between tissues, this relates to the functions of the RAD as growing tissue and the CAP as a regulator of germination through weakening. More detailed analysis showed that putative orthologs of cell wall-remodeling genes are expressed in a complex manner during CAP weakening, suggesting distinct roles in the RAD and CAP. Expression patterns are also consistent with the CAP being a receptor for environmental signals influencing germination. Inhibitors of the aspartic, serine, and cysteine proteases reduced the number of isolated CAPs in which weakening developed, and inhibition of the 26S proteasome resulted in its complete cessation. This indicates that targeted protein degradation is a major control point for endosperm weakening.

Aerobic treatment of winery wastewater with the aim of water reuse.

An air micro-bubble bioreactor (AMBB) using a free self-adapted microbial population, 15 dm3 working volume, was used for aerobic treatment of winery wastewater. This reactor utilizes a Venturi injector in conjunction with mass transfer multiplier nozzles, which allow an efficient oxygen transfer. The reactor can operate in batch or continuous conditions. The dynamics of chemical oxygen demand (COD), biomass and total contents of polyphenolic compounds was followed throughout each trial. The wastewater COD ranged between 4.0-8.0 kg COD m(-3) and the efficiency of the batch treatment was about 90.0 +/- 4.3%, after 6 days of operation. The maximum efficiency obtained was achieved after 15 days of treatment (99%). In continuous conditions, the loading rate and the treatment efficiency ranged between 0.45-1.00 kg COD m(-3) d(-1) and 93.3 +/- 2.0%, respectively. The AMBB hydraulic retention time was 15 days. To assess the suitability of treated water in relation to vineyard irrigation, the effluent was physico-chemical analysed and direct toxicity bioassays with effluent matrix were carried out using Lepidium sativum L. seeds. The results showed the water quality required to be reutilised minimizing water consumption. This study will contribute for the implementation of an efficient water use plan, aiming the preservation of the water resource and the reduction of the wastewater production.

Transport of cadmium and assessment of phytotoxicity after electrokinetic remediation.

The use of ethylene diamine tetraacetic acid (EDTA) on the electrokinetic removal of cadmium-contaminated soil was evaluated. A total of four different tests were conducted using EDTA as a washing solution as well as a purging solution at the electrode compartments. The efficiency of electrokinetic extraction was significantly influenced by the pH of the soil medium. The results show that EDTA was effective in desorbing cadmium at a high pH, with Cd-EDTA(-) anion complexes migrating toward the anode. At low pH values near the anode area, cadmium existed as Cd(2+), migrating toward the cathode. Such contradicting directions of cadmium have resulted in its detrimental removal from the soil cell. However, accumulation of cadmium near the cathode was observed at the end of the tests due to the dominating low pH in the soil cell. The phytotoxicity after the electrokinetic process was investigated using Sorghum saccharatum, Lepidium sativum and Sinapis alba plants. The germination index was a major endpoint estimated by measuring seed germination and shoot elongation. The results obtained show that the phytotoxicity was increased after electrokinetic extraction. Despite, the extensive cadmium removal from two-thirds of the cell, the low pH of the soil was the principal parameter exhibiting the phytotoxicity.

Land spreading of olive mill wastewater: effects on soil microbial activity and potential phytotoxicity.

Extremely high organic load and the toxic nature of olive mill wastewater (OMW) prevent their direct discharge into domestic wastewater treatment systems. In addition to the various treatment schemes designed for such wastewater, controlled land spreading of untreated OMW has been suggested as an alternative mean of disposal. A field study was conducted between October 2004 and September 2005 to assess possible effects of OMW on soil microbial activity and potential phytotoxicity. The experiment was carried out in an organic orchard located on a Vertisol-type soil (Jezre'el Valley, Israel) and included two application levels of OMW (36 and 72m(3)ha(-1)). Total microbial counts, and to less extent the hydrolytic activity and soil respiration were increased following the high OMW application level. A bench-scale lab experiment showed that the rate of OMW mineralization was mainly dependent on the general status of soil activity and was not related to previous acclimatization of the soil microflora to OMW. Soil phytotoxicity (% germination and root elongation) was assessed in soil extracts of samples collected before and after each OMW application, using germinating cress (Lepidium sativum L.) seeds. We found direct short-term effect of OMW application on soil phytotoxicity. However, the soil was partly or completely recovered between successive applications. No further phytotoxicity was observed in treated soils as compared with control soil, 3 months after OMW application. Such short-term phytotoxicity was not in correlation with measured EC and total polyphenols in the soil extracts. Overall, the results of this study further support a safe controlled OMW spreading on lands that are not associated with sensitive aquifers.

Comparative study on sensitivity of higher plants and fish to heavy fuel oil.

Laboratory tests were conducted on higher plants [garden cress (Lepidium sativum), great duckweed (Spirodela polyrrhiza), and Tradescantia clone BNL 02] and fish [rainbow trout (Oncorhynchus mykiss) at all stages of development: eggs, larvae and adults] to estimate their sensitivity to heavy fuel oil (HFO). A number of biological indices (survival, growth, and physiological and morphological parameters) as well as the genotoxic impact (Tradescantia) of HFO was evaluated by acute and chronic toxicity tests. Fish were found to be more sensitive to the toxic effect of HFO than were higher plants. EC(50) values obtained for higher plants ranged from 8.7 g/L (L. sativum) to 19.8 g/L (Tradescantia), and maximum-acceptable-toxicant concentration (MATC) values ranged from 0.1 to 1.0 g/L of total HFO for L. sativum and Tradescantia, respectively. The 96-h LC(50) values ranged from 0.33 g/L, for larvae, to 2.97 g/L, for adult fish, and the MATC value for fish was found to be equal to 0.0042 g/L of total HFO. To evaluate and predict the ecological risk of the overall effects of oil spills, studies should be performed using a set of acute and chronic bioassays that include test species of different phylogenetic levels with the most sensitive morphological, physiological, and genotoxic indices.