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A simple and efficient method for testing Lettuce mosaic virus resistance in in vitro cultivated lettuce.

The potential of a new in vitro inoculation and propagation method developed for Lettuce mosaic virus (LMV) on lettuce (Lactuca sativa L.) was evaluated by studying LMV infection on in vitro cultivated seedlings or on newly regenerated plantlets. Lettuce cultivars differing by their LMV-resistance status were inoculated with various natural LMV isolates as well as with Green Fluorescent Protein (GFP)-tagged recombinant virus isolates. A good correlation was observed between the known resistance status of the cultivars and the results obtained by in vitro screening. The results show that this resistance assay can be greatly improved by the use of GFP-tagged virus isolates. The main advantages of this method are reduced space requirements and an improved environmental safety, especially for the handling of recombinant virus, of quarantine virus or of transgenic plants.

Enzyme-Linked Immunosorbent Assay↗

Structure elucidation and phytotoxicity of C13 nor-isoprenoids from Cestrum parqui.

Twelve C(13) nor-isoprenoids have been isolated from the leaves of Cestrum parqui (Solanaceae). The structure (2R,6R,9R)-2,9-dihydroxy-4-megastigmen-3-one has been assigned to the new compound. All the structures have been determined by spectroscopic means and chemical correlations. The compounds showed phytotoxic effect on the germination and growth of Lactuca sativa L.

Cestrum↗

Arudonine, an allelopathic steroidal glycoalkaloid from the root bark of Solanum arundo Mattei.

Bioassay-guided fractionation of the methanol extract of the root bark of Solanum arundo Mattei led to the isolation of a steroidal glycoalkaloid, designated arudonine, which was elucidated to be solasodine O-alpha-l-rhamnopyranosyl-(1-->2)-[beta-D-xylopyranosyl-(1-->3)], [alpha-l-rhamnopyranosyl-(1-->4)]-beta-D-glucopyranoside. This steroidal glycoalkaloid inhibited the growth of lettuce seedlings (Lactuca sativa).

Biological Assay↗

Terpenoids and phenol derivatives from Malva silvestris.

A sesquiterpene and a tetrahydroxylated acyclic diterpene as well as two known monoterpenes, 6 C(13)nor-terpenes and 11 aromatic compounds were isolated from the water extract of Malva silvestris. The structures of the compounds were determined by spectroscopic NMR and MS analysis. Effects of these compounds on germination and growth of dicotyledon Lactuca sativa L. (lettuce) were studied in the 10(-4)-10(-7)M concentration range.

Lactuca↗

Structures of bioactive carexanes from the roots of Carex distachya Desf.

Four metabolites, named carexanes I-L, have been isolated from the roots of Carex distachya Desf, an herbaceous plant living in the Mediterranean maquis, together with three known compounds, already isolated from the aerial part of the plant. All the compounds have been characterized on the basis of their spectroscopic properties. Carexane I derived from the lose of a proton from the C-18 carbon of an intermediate isopropyl cation. Its stereostructure has been elucidated by Mosher's method, NOESY/ROESY experiments and computational calculations. The bioactivity on seed germination and root/shoot growth of Lactuca sativa L. of all the isolated compounds is also reported.

Carex Plant↗

Response to ozone in two lettuce varieties on chlorophyll a fluorescence, photosynthetic pigments and lipid peroxidation.

The effect of different O3 concentrations on two lettuce (Lactuca sativa L.) varieties (Valladolid and Morella) was investigated through chlorophyll (Chl) a fluorescence parameters, photosynthetic pigments (Chl a, b and total carotenoid), lipid peroxidation and crop yield. Ozone fumigation caused: a decrease in maximum quantum yield of photosystem II (PSII) photochemistry (Fv/Fm) in mature leaves, a reduction in the non-cyclic electron flow (phiPSII) and a lower capacity to reoxidize the QA pool (qP). These reductions were significant in the Valladolid var. but not in the Morella var. A significant decrease in Chl a, b and in the total carotenoids was observed in the Valladolid var. but not in the Morella var. mainly under O3 fumigation conditions. We observed that the NPQ parameter did not increase in parallel to the qP reduction seen in the Valladolid var. O3 fumigation with respect to air charcoal filtered air conditions. This fact could be associated with a lower capacity for dissipation of non-radiative excess energy and it may be closely correlated with significant decreases in photosynthetic pigment concentration. A decrease in NPQ from air ozone-free to ozone fumigation in the Morella var. can be explained by the need to maintain the photochemical quenching under O3 stress. It may also be associated with a slight increase in photosynthetic pigments. The differences between the two varieties may indicate that the Valladolid var. is more susceptible to O3 damage.

Chlorophyll↗

Extractability of metals and ecotoxicity of soils from two old wood impregnation sites in Finland.

Four metal-contaminated soil samples were classified using physical methods, extracted by selective extraction procedures and analyzed for chemical concentrations. De-ionized water, 0.01 mol/l barium chloride, 1 mol/l ammonium acetate and concentrated nitric acid were used as extraction solutions. Ecotoxicity of water extracts and soil samples was analyzed in order to describe the bioavailability of the contaminants. Samples from old wood impregnation plants contained high amounts of As, Cu, Cr and Zn, which originated from chromated copper arsenate, ammoniacal copper-zinc arsenate, and ammoniacal copper quaternary compound. Total As concentrations of the heavily contaminated samples varied from 752 to 4340 mg/kg, Cu concentrations from 339 to 2330 mg/kg, Cr concentrations from 367 to 2,140 mg/kg and Zn concentrations from 79 to 966 mg/kg. The extractabilities of metals differed according to soil type, extractant and element. Cu and Zn were proposed to cause the highest toxicity in the water extracts of the soils. Ecotoxicity tests displayed rather high differences in sensitivity both for water extracts and for solid soil samples. Reproduction of Enchytraeus sp. was the most sensitive and seed germination of Lactuca sativa the least sensitive and the other tests were in decreasing order of sensitivity: Folsomia candida>reverse electron transport>MetPLATE>Toxichromotest>Allium cepa root growth>Lemna sp. growth. As a conclusion, polluted soils rich in sand retain heavy metals with less firm bindings, particularly in the case of Cu and Zn, than soils rich in clay, indicating that chemical methods for measuring the bioavailability of metals need to be optimized taking into account the soil type, acidity, redox state and the individual contaminants.

Allium↗

Cadmium phytotoxicity: Quantitative sensitivity relationships between classical endpoints and antioxidative enzyme biomarkers.

In this work, cadmium phytotoxicity and quantitative sensitivity relationships between different hierarchical endpoints in plants cultivated in a contaminated soil were studied. Thus, germination rate, biomass growth and antioxidative enzyme activity (i.e. superoxide dismutase, peroxidase, catalase and glutathione reductase) in three terrestrial plants (Avena sativa L., Brassica campestris L. cv. Chinensis, Lactuca sativa L. cv. hanson) were analyzed. Plant growth tests were carried out according to an International Standard Organization method and the results were analyzed by ANOVA followed by Williams' test. The concentration of Cd2+ that had the smallest observed significant negative effect (LOEC) on plant biomass was 6.25, 12.5 and 50 mg Cd/kg dry soil for lettuce, oat and Chinese cabbage, respectively. Activity of all enzymes studied increased significantly compared to enzyme activity in plant controls. For lettuce, LOEC values (mg Cd/kg dry soil) for enzymic activity ranged from 0.05 (glutathione reductase) to 0.39 (catalase). For oat, LOEC values (mg Cd/kg dry soil) ranged from 0.19 (for superoxide dismutase and glutathione reductase) to 0.39 (for catalase and peroxidase). For Chinese cabbage, LOEC values (mg Cd/kg dry soil) ranged from 0.19 (peroxidase, catalase and glutathione reductase) to 0.39 (superoxide dismutase). Classical (i.e. germination and biomass) and biochemical (i.e. enzyme activity) endpoints were compared to establish a sensitivity ranking, which was: enzyme activity>biomass>germination rate. For cadmium-soil contamination, the determination of quantitative sensitivity relationships (QSR) between classical and antioxidative enzyme biomarkers showed that the most sensitive plant species have, generally, the lowest QSR values.

Antioxidants↗

Effect of scale of Cd heterogeneity and timing of exposure on the Cd uptake and shoot biomass, of plants with a contrasting root morphology.

A pot experiment was conducted to investigate the influence of spatial heterogeneity of Cd distribution in soil on shoot biomass, shoot metal concentration and total shoot Cd uptake by lettuce (Lactuca sativa, variety Tom Thumb) and Indian mustard (Brassica juncea). Five different soil treatments had similar overall concentration of Cd per pot, but different scales of heterogeneity and also timing of plant exposure during the growth cycle. The presence and scale of heterogeneity and timing of exposure were found to have significant effects on shoot biomass for both plants (with one exception). The mean values of Cd mass taken up were significantly affected by the presence of heterogeneity and timing only for lettuce. Only the scale of heterogeneity affected the uptake of Cd by Indian mustard, presumably because of its larger root system (approximately 18 cm, compared with approximately 5 cm for lettuce). These findings have important implications for phytoremediation, and for human health risk assessment where leafy vegetables are grown in situations with highly elevated Cd concentrations.

Analysis of Variance↗

Cadmium assimilation in the terrestrial isopod, Porcellio dilatatus--is trophic transfer important?

Terrestrial isopods have become important tools for the ecotoxicological assessment of metal-contaminated soils. Their value as an invertebrate model is partly because of their extraordinary capacity to bioaccumulate toxic metals from the environment. Replication of this accumulation process in the laboratory has in the past relied on the amendment of organic food substrates through the addition of inorganic metal salts. However, the bioavailability of the metals when presented through doping regimes may differ from the bioavailability of metals in nature, because over time metals become biologically compartmentalised and form complexes with organic molecules. This study examines the differential bioavailability of Cd to the terrestrial isopod, Porcellio dilatatus, when presented as either a Cd-amended diet or pre-incorporated biologically into lettuce (Lactuca sativa). Isopods were either provided with lettuce contaminated superficially with Cd(NO(3))(2) or lettuce grown hydroponically in growth media containing 100 microM Cd(NO(3))(2). Assimilation efficiency of Cd was greater among isopods that were fed the amended diet (71%, S.E.=7%), than among isopods feeding on biologically contaminated lettuce (52%, S.E.=5%) and demonstrates that speciation of Cd is likely to influence the rate of Cd assimilation and accumulation in a laboratory test.

Animal Feed↗

Isolation and identification of an allelopathic substance from peel of Citrus junos.

The inhibitory effect of Citrus junos peel on plant growth using lettuce (Lactuca sativa L.) as a bioassay material was investigated, since the powder of the peel had been found to inhibit growth of weeds. Basic, neutral and acidic fractions were separated from the aqueous fraction obtained from the methanol extract of C. junos peel. All fractions inhibited the growth of lettuce seedlings, but by far the greatest inhibition was observed with the neutral fraction. Thus, the latter was further purified and an allelopathically active substance was isolated. The structure of the substance was determined from high-resolution MS and 1H and 13C NMR spectral data as abscisic acid-beta-D-glucopyranosyl ester (ABA-GE). ABA-GE inhibited hypocotyl and root growth of lettuce seedlings at concentrations greater than 0.3 microM, and the concentrations for 50% inhibition of hypocotyl and root growth were 2.3 and 1.4 microM, respectively. The effectiveness of ABA-GE on inhibition of growth and the occurrence of ABA-GE in the peel itself suggested that ABA-GE may play an important role in the allelopathic potential of C. junos peel. The peel may be potentially useful for weed management in a field setting.

Abscisic Acid↗

Isolation and identification of an allelopathic substance in Pisum sativum.

The residue of peas (Pisum sativum L.) has allelopathic activity and the putative compound causing this inhibitory effect was isolated from a methanol extract of pea shoots. Chemical structure of this compound was determined by high-resolution MS, IR and 1H NMR spectral data as pisatin. Pisatin inhibited growth of cress (Lepidium sativum L.) and lettuce (Lactuca sativa L.) seedlings at concentrations greater than 10 and 30 microM, respectively. The doses required for 50% growth inhibition of roots and hypocotyls of cress were 61 and 91 microM, respectively, and those of lettuce were 78 and 115 microM, respectively. The concentration of pisatin in the pea shoots was 32.7 nmol x g(-1) fresh weight. The effectiveness of pisatin on growth inhibition in cress and lettuce, and its occurrence in pea shoots suggest that it may contribute to the growth inhibitory effect of pea residue, and may play an important role in pea allelopathy.

Chromatography, High Pressure Liquid↗

Transfer of the fungicide vinclozolin from treated to untreated plants via volatilization.

Head lettuce plantlets (Lactuca sativa L. var. capitata) were potted, treated with vinclozolin at the six-leaf stage according to application standards and allowed to dry for 24 h. The potted plantlets were then placed in either growth chambers with controlled temperature (20 and 25 degrees C, respectively) or in a greenhouse (approximately 12 degrees C), together with untreated spinach (Spinacia oleracea L.) and standardized grass cultures (Lolium multiflorum Lam. ssp.) While the treated lettuce pots remained in the respective growing compartments until the end of the experiments, spinach and grass were exposed to the compartment air for 24 h and their shoot material was analyzed for vinclozolin by GC-ECD and GC-high resolution mass spectrometry. Exposure and analysis of untreated spinach and grass were carried out at two- or three-day intervals during the course of the experiments. Also, air samples were taken from the compartments at intervals and analyzed for vinclozolin. Maximum vinclozolin concentration in the growth chamber air was about 330 ng m(-3) while vinclozolin contamination of the untreated plants ranged from 50 to 200 microg kg(-1) FW (fresh weight). In the greenhouse atmospheric vinclozolin concentration reached approximately 15 ngm(-3) and maximum contamination of spinach and grass were 30-40 microg kg(-1) FW. Our data clearly show that unintended contamination of plants growing in the vicinity of vinclozolin-treated plants can occur even if the fungicide layer is completely dry. Implications for safety testing and food plants are discussed.

Air Pollutants↗

Uptake of perchlorate in terrestrial plants.

Cucumber (Cucumis sativus L.), lettuce (Lactuca sativa L.), and soybean (Glycine max) were used to determine uptake of the perchlorate anion (100 ppb) from sand. Plants were watered with different ratios of Hydrosol (a diluted solution of Peters All-Purpose Plant Food) to Milli-Q water (18 MOmega) to determine if the presence of other nutrients (such as nitrate) influenced perchlorate uptake. Perchlorate concentrations in sand and plant tissues were determined weekly. Perchlorate uptake was observed in all three plant species. In most experiments, perchlorate was completely depleted from sand in which plants were growing. Perchlorate concentrations in lettuce were also significantly higher than those in cucumber and soybean (P < 0.0001). Perchlorate concentrations in sand decreased at a higher rate at lower ratios of Hydrosol to Milli-Q, indicating that plant (cucumber) uptake of perchlorate is influenced by the presence of external nutrients. The results of an 8-week uptake study in cucumber and a 6-week uptake study in lettuce suggest that a threshold perchlorate concentration is reached: for cucumber, 150 ppm and for lettuce, 750 ppm. Although the presence of external nutrients decreases the rate of perchlorate uptake by plants, significant concentrations of perchlorate occur in aboveground plant tissues even after relatively short periods of growth. The potential for trophic transfer of perchlorate from soil to higher organisms through plants exists.

Biological Availability↗

Effects of Se and Zn supplementation on the antagonism against Pb and Cd in vegetables.

The antagonistic effects of supplementation of Zn and Se to the soil on vegetables were studied in this work. In the pot experiment, Se (Se4+) and Zn (Zn2+) were applied, respectively, to the soil, in which the Chinese cabbage (Brassica rapa) and the lettuce (Lactuca sativa L.) were planted. As a result, Se and Zn were enriched evidently in the two vegetables. The contents of Pb and Cd in the two vegetables were decreased markedly while contents of some healthy mineral elements, like Mn and Mg, were increased to some extent when Se and Zn were applied. The antagonism of Se and Zn against Pb and Cd in plants was suggested. The farmland experiment on the lettuce was conducted to explore further the effect of supplementation of Zn and Se under the actual field conditions. Result came out to be that the enrichment of Zn and Se restrained the accumulation of Pb and Cd in the lettuce remarkably, as well as enhanced the absorption of some other nutritional elements, like Fe, Mn, Cu, Ca and Mg. Therefore, application of Se and Zn was proved to be an effective and feasible method to improve trace elements nutrition in the vegetables.

Brassica↗

Ontogeny of plants under various gravity condition.

The results of experiments performed under conditions of microgravity (MG) or under its simulation on the horizontal clinostat (HC) with the callus, seedlings of various species and embryogenic structures have revealed a definite role of gravity as an ecological factor in the processes of cytomorphogenesis, growth, and development. The transformation of differentiated somatic cells of arabidopsis seed into undifferentiated callus was not inhibited under MG, though modifications of the whole callus morphology and of mean cell and nucleus size were observed. The morphogenesis of polar structures such as root-hair bearing cells of Lactuca primary root has been shown to be modified in the course of differentiation under mass acceleration diminished below 0.1 g. Seed germination and seedling morphogenesis under MG follow their normal course, but a significant stimulation of shoot growth with no effect on primary root growth has been determined. A successful in vitro regeneration of Nicotiana tabacum plantlets from leaf cells and subsequent formation of shoots and roots on a continuously rotating HC as well as the formation of viable seeds during seed-to-seed growth of Arabidopsis plants under MG have indicated that gravity plays but a limited role in the processes of embryogenesis and organogenesis.

Arabidopsis↗

Integrating biological treatment of crop residue into a hydroponic sweetpotato culture.

Residual biomass from hydroponic culture of sweetpotato [Ipomoea batatas (L.) Lam.] was degraded using natural bacterial soil isolates. Sweetpotato was grown for 120 days in hydroponic culture with a nutrient solution comprised of a ratio of 80% modified half Hoagland solution to 20% filtered effluent from an aerobic starch hydrolysis bioreactor. The phytotoxicity of the effluent was assayed with Waldmann's Green' lettuce (Lactuca sativa L.) and the ratio selected after a 60-day bioassay using sweetpotato plants propagated vegetatively from cuttings. Controlled environment chamber experiments were conducted to investigate the impact of filtrate from biological treatment of crop residue on growth and storage root production with plants grown in a modified half Hoagland solution. Incorporation of bioreactor effluent, reduced storage root yield of 'Georgia Jet' sweetpotato but the decrease was not statistically significant when compared with yield for plants cultured in a modified half Hoagland solution without filtrate. However, yield of 'TU-82-155' sweetpotato was significantly reduced when grown in a modified half Hoagland solution into which filtered effluent had been incorporated. Total biomass was significantly reduced for both sweetpotato cultivars when grown in bioreactor effluent. The leaf area and dry matter accumulation were significantly (P < 0.05) reduced for both cultivars when grown in solution culture containing 20% filtered effluent.

Biodegradation, Environmental↗

An ecotoxic risk assessment of residue materials produced by the plasma pyrolysis/vitrification (PP/V) process.

Plasma is the fourth state of matter, following the three states of solid, liquid and gas. Experience has amply demonstrated that solids exposed to the oxygen-deficient plasma flame are converted to liquid, and liquid exposed to the same flame is converted to gas. A low amount of vitrified solid residue material usually remains at the end of this process. Plasma pyrolysis/vitrification (PP/V) has been demonstrated as a safe, efficient, cost-effective technology for the treatment of wastes, including hazardous wastes. Besides the low amounts of gaseous byproducts that PP/V produces, the solid vitrified residue presents a low leachability of pollutants. Studies have been performed in many countries in order to assess the leachability of chemical substances. But from the results of identified studies, none has reported results on the ecotoxicological properties of the leachates. The aim of this study was to contribute to the assessment of ecotoxic risk of four different vitrified materials. Vitrified samples of contaminated soils, municipal solid wastes, and incinerator bottom ashes were submitted to the European leaching pre-standard test number prEN 12457-2. The leachates were analyzed for 22 chemical parameters. The biological characterization comprised the assessment of bioluminescence inhibition of Photobacterium phosphoreum bacterium, growth inhibition of Pseudokirchneriella subcapitata algae and the germination inhibition of Lactuca sativa vegetable. The chemical and ecotoxicological results were analyzed according to the French proposal of Criteria on the Evaluation Methods of Waste Toxicity (CEMWT) and a Toxicity Classification System (TCS). The chemical and ecotoxicological results indicated a low leachability of pollutants and a low toxicity level of leachates. All samples studied were as below the TCS class 1 level (no significant toxicity observed) and as non-ecotoxic for CEMWT. Therefore, the environmental ecotoxic risk of the analyzed vitrified samples was determined to be very low.

Chemical Phenomena↗