Acyl-homoserine lactones produced by Pantoea sp. isolated from the "maize white spot" foliar disease

The "maize white spot" foliar disease is a problem of increasing importance to Brazilian maize crops. A bacterium isolated from water-soaked lesions from infected maize leaves was pathogenic in biological assays in vivo. It was identified as a Gram-negative, nonsporulating, facultative anaerobic bacterium, belonging to the genus Pantoea. Chemical study of the extracts from bacterial cultivation media allowed the identification of (S)-(-)-N-butanoyl-homoserine lactone and trace amounts of N-hexanoyl-homoserine lactone, widely recognized quorum-sensing signaling substances employed in cell-to-cell communication systems. The absolute configuration of natural (S)-(-)-N-butanoyl-homoserine lactone was determined by gas chromatography-flame ionization detection with a chiral stationary phase and by comparison of circular dichroism spectroscopic data with enantiopure synthetic substances. Biological evaluations with reporter Agrobacterium tumefaciens NTL4(pZLR4) were carried out with synthetic and natural products and also with extracts from maize leaves contaminated with the isolated bacterium, as well as from healthy leaves.     

Soybean (Glycine max) Extracts Impact on Plant and Soil Biology

The aim of the present investigation was to determine the phytotoxic effect of aqueous extract from fresh and oven-dried leaves and roots of soybean plants on maize growth and on rhizosphere soil. The extract prepared from fresh leaves of drought-stressed soybean plants significantly increased the accumulation of proline, soluble sugar, and endogenous abscisic acid (ABA) content of maize-treated plants. Increase in antioxidant activity like superoxide dismutase (SOD), peroxidase (POD), ascorbate peroxidase (APX), catalase (CAT), and endogenous ABA was recorded in treated plants. The soybean leaf extracts prepared from drought-stressed plants significantly decreased the soil pH and electrical conductivity (EC) of maize cultivated soil. The soybean drought-stressed leaf extracts prepared plants were found more effective in modulating the physiology of maize, indicating the higher allelopathic potential of soybean.     

Microwave-assisted efficient extraction of different parts of Hippophae rhamnoides for the comparative evaluation of antioxidant activity and quantification of its phenolic constituents by reverse-phase high-performance liquid chromatography (RP-HPLC)

The outcome of different extraction procedures (microwave, ultrasound, Soxhlet, and maceration) on the antioxidant activity of seeds, leaves, pulp, and fruits of Hippophae rhamnoides (sea buckthorn or SBT) was investigated by two different bioassays: 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) and 2,2'-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assays. The SBT extracts were found to possess strong antioxidant activity measured in terms of TEAC (2.03-182.13 and 6.97-282.75 mg/g) with ABTS and DPPH assays, respectively. In general, the antioxidant capacity of microwave-assisted extracts was found to be significantly higher than those obtained by ultrasound-assisted extraction (UAE) and maceration while being slightly higher than Soxhlet extracts. Further, microwave extracts of seeds were found to possess maximum antioxidant capacity followed by leaves, fruits, and pulp. Also, the chemical composition of extracts, studied in terms of the total phenolic content, was found to be in the range of 1.9-23.5 mg/g Gallic acid equivalent (GAE), which indicates a strong correlation between antioxidant activity and phenolic content present in the SBT. In addition, some of its bioactive phenolic constituents, such as rutin ( 1), quercetin-3- O-galactoside ( 2), quercetin ( 3), myricetin ( 4), kaempferol ( 5), and isorhamnetin ( 6), were also quantified in different extracts by reverse-phase high-performance liquid chromatography (RP-HPLC).     

根系互作对玉米大豆间作作物磷吸收的影响

【目的】间作作物的养分吸收与根系相互作用有着密切关系。研究玉米大豆间作后根系互作对磷含量、磷积累动态的影响,旨在为以玉米大豆为主的间作体系中磷的高效利用提供科学依据。【方法】本文通过盆栽试验,采用玉米和大豆根系无分隔(NB)、尼龙网分隔(MB)、塑料膜分隔(PB)3种分隔方式,研究了间作玉米(苗期、大喇叭口期、孕穗期、成熟期)和大豆(苗期、分枝期、鼓粒期、成熟期)在不同生育期对土壤中磷的吸收。【结果】与PB相比,MB处理的玉米茎中磷含量在孕穗期提高了16.8%(P0.05);叶中磷含量在苗期、大喇叭口期分别提高了6.6%、14.2%(P0.05);NB处理玉米茎中磷含量在大喇叭口期、孕穗期分别提高了25.4%、55.0%(P0.05);玉米叶中磷含量在大喇叭口期、成熟期分别提高了21.3%、22.2%(P0.05);与PB相比,NB处理大豆茎中磷含量在分枝期、成熟期分别提高了21.3%、24.7%(P0.05);叶中磷含量在分枝期提高了11.4%(P0.05),MB处理大豆叶中磷含量在分枝期提高了4.8%(P0.05)。与PB相比,NB处理玉米茎中磷积累量在大喇叭口期、孕穗期提高了24.6%、32.3%(P0.05),MB处理玉米茎中磷积累量在大喇叭口期提高了50.6%(P0.05),叶中磷积累量在苗期提高了33.6%(P0.05)。与PB相比,NB处理大豆茎中磷积累量在分枝期、鼓粒期分别提高了36.3%、51.8%(P0.05),叶中磷积累量在分枝期、成熟期分别提高了27.3%、110.6%(P0.05);MB处理大豆茎中磷积累量在鼓粒期提高了35.7%(P0.05)。【结论】根系互作系统中的玉米和大豆茎、叶中磷的累积量大于根系分离生长系统。因此,玉米大豆间作具有明显的磷吸收利用优势。     

黄顶菊水提液对玉米幼苗株高及叶片生理生化指标的影响

以黄顶菊新鲜植株为材料,用蒸馏水浸提活性物质,得到0.1、0.05和0.01g/mL三种不同干物质含量的黄顶菊水提液.以蒸馏水为对照,利用不同含量的黄顶菊水提液处理无土栽培的玉米幼苗,每4d测定幼苗叶片中MDA(丙二醛)、POD(过氧化物酶)、SOD(超氧化歧化酶)、CAT(过氧化氢酶)和CHL(叶绿素)含量.结果表明,黄顶菊水提液处理对玉米幼苗株高产生了明显的抑制作用,且每个处理与对照相比差异达显著水平(P＜0.05).黄顶菊水提液中含量越高,抑制作用越显著.幼苗叶片中MDA含量、POD、SOD、CAT活性均高于对照,而CHL含量低于对照.研究还发现,不同含量处理和不同处理时间对玉米幼苗MDA含量、抗氧化酶和CHL含量的影响之间存在极显著的交互作用.结果预示黄顶菊水提液能够抑制幼苗生长,含量越大,作用效果越明显,可为进一步研究黄顶菊总体化感提供参考.     

Rapid quantification of proanthocyanidins (condensed tannins) with a continuous flow analyzer

Proanthocyanidins (condensed tannins) frequently need to be quantified in large numbers of samples in food, plant, and environmental studies. An automated colorimetric method to quantify proanthocyanidins with sulfuric acid (H(2)SO(4)) was therefore developed for use in a continuous flow analyzer. Assay conditions were optimized using 50% methanol extracts of paper birch, sugar maple, and quaking aspen leaves. Short extraction times and centrifugation of samples prevented proanthocyanidin degradation that otherwise occurred in 50% methanol extracts of aspen leaves. Extraction of birch and maple proanthocyanidins with 50% methanol was comparable to or better than that with 70% acetone. Proanthocyanidin levels in aspen were lower when extracted with aqueous methanol, but relative differences among samples were consistent with those found in aqueous acetone extracts. Results from the automated sulfuric acid assay were highly correlated with those of the conventional BuOH-HCl method for proanthocyanidins and, except for birch, with the Folin--Denis assay for total phenolics. This new technique significantly improves assay processing rate and repeatability compared to conventional colorimetric proanthocyanidin assays.     

Inhibitory effects of feeding with carrots or (-)-falcarinol on development of azoxymethane-induced preneoplastic lesions in the rat colon

The effects of intake of dietary amounts of carrot or corresponding amounts of (-)-(3R)-falcarinol from carrots on development of azoxymethane (AOM)-induced colon preneoplastic lesions were examined in male BDIX rats. Three groups of eight AOM-treated rats were fed the standard rat feed Altromin supplemented with either 10% (w/w) freeze-dried carrots with a natural content of 35 mug falcarinol/g, 10% maize starch to which was added 35 mug falcarinol/g purified from carrots, or 10% maize starch (control). After 18 weeks, the animals were euthanized and the colon was examined for tumors and aberrant crypt foci (ACF), which were classified into four size classes. Although the number of small ACF was unaffected by the feeding treatments, the numbers of lesions as a function of increasing size class decreased significantly in the rats that received one of the two experimental treatments, as compared with the control treatment. This indicates that the dietary treatments with carrot and falcarinol delayed or retarded the development of large ACF and tumors. The present study provides a new perspective on the known epidemiological associations between high intake of carrots and reduced incidence of cancers.     

Metabolism of fenitrothion and conjugation of 3-methyl-4-nitrophenol in tomato plant (Lycopersicon esculentum)

The metabolism of (14)C-labeled fenitrothion (Sumithion, [O,O-dimethyl-O-(3-methyl-4-nitrophenyl)phosphorothioate]) in tomato plant (Lycopersicon esculentum Mill., cv. Ponderosa) grown in the greenhouse equipped with quartz glass was conducted to investigate the effect of sunlight on the behavior of fenitrothion and to elucidate the detailed structure of conjugated metabolites. Tomato plants (BBCH 85) were topically treated with (14)C-labeled fenitrothion twice with a 2 week interval between applications. At 15 days after the second application, more than half of the recovered (14)C was detected as unaltered fenitrothion, glucose, and cellobiose esters of 3-methyl-4-nitrophenol (NMC) in extracts from tomato fruit. The photoinduced formation of the S-methyl isomer of fenitrothion via thiono-thiolo rearrangement was detected only in the surface rinse but at trace amounts. In the whole tomato fruit, fenitrothion, the S isomer, NMC-beta-glucoside, and NMC cellobioside were detected at 34.16, 1.28, 7.47, and 15.07% of the recovered (14)C, respectively. Trace amounts of the oxon analogue of fenitrothion were detected only on tomato leaves. The chemical structure of the cellobiose conjugate of NMC, 1-O-beta-d-glucopyranosyl-(1-->4)-beta-d-glucopyranosyl-3-methyl-4-nitrophenol, was determined by spectroscopic analyses (liquid chromatography-mass spectrometry, NMR), using the metabolite obtained from leaves and stems of tomato plant hydroponically grown with (14)C-labeled NMC.     

Volatile components and key odorants of fennel (Foeniculum vulgare Mill.) and thyme (Thymus vulgaris L.) oil extracts obtained by simultaneous distillation-extraction and supercritical fluid extraction

Volatile oil extracts of fennel seeds (Foeniculum vulgare Mill.) and thyme leaves (Thymus vulgaris L.) were obtained by simultaneous distillation-extraction (SDE) and supercritical fluid extraction (SFE) and analyzed by gas chromatography-mass spectrometry (GC-MS). In general, fennel oil extracted by SDE and SFE showed similar compositions, with trans-anethole, estragole, and fenchone as the main components. In contrast, thymol and p-cymene, the most abundant compounds in thyme leaves, showed big differences, with generally higher amounts of monoterpenes obtained by SDE. However, in this case, the differences between the extracts were higher. Key odorants of fennel seeds determined by gas chromatography-olfactometry (GC-O) showed similar patterns when applying SDE and SFE. trans-Anethole (anise, licorice), estragole (anise, licorice, sweet), fenchone (mint, camphor, warm), and 1-octen-3-ol (mushroom) were the most intense odor compounds detected in fennel extracts. Thymol and carvacrol, with oregano, thyme, and spicy notes, were identified as key compounds contributing to the aroma of thyme leaves.     

Immunostimulating activity of a crude polysaccharide derived from green tea (Camellia sinensis) extract

Green tea extract is well-known to reduce the risk of a variety of diseases. Here, we investigated the immunostimulating activity of tea polysaccharide (TPS), one of the main components in green tea extract. The water extracts from mature or immature tea leaves were precipitated by using ethanol at room temperature. The sediment was washed with ethanol and acetone alternately and then dried. We used the phagocytic activity of macrophage-like cells as an indicator of immune function activation. Chemical components were analyzed by HPLC. The immunostimulating activity of TPS from immature leaves extract was higher than that of TPS from mature leaves, and its activities were dependent on the content of strictinin in the leaf extract. Futhermore, a mixture of catechin and TPS that removed polyphenols did not increase the immunostimulating activity. These results suggest that the catechin-polysaccharide complex is a very important molecule in the immunomodulating activity of tea extracts.     

Determination of tea polyphenols and caffeine in tea flowers (Camellia sinensis) and their hydroxyl radical scavenging and nitric oxide suppressing effects

The native occurrence of tea polyphenols, namely, (-)-epicatechin, (+)-catechin, (-)-epigallocatechin 3-gallate, (-)-epicatechin, and (-)-epicatechin 3-gallate, and caffeine in tea flowers was assessed by an isocratic HPLC procedure. The levels of total catechins and caffeine were determined in tea flowers collected from 10 different species of Camellia sinensis. The results showed the levels of total catechin ranged from 10 to 38 mg/g, whereas the level of caffeine ranged from 3 to 8 mg/g. Levels of catechins and caffeine in tea leaves and various teas were also determined and ranged from 2 to 126 mg/g and from 23 to 49 mg/g, respectively. Both tea flower and tea leaf extracts exert their strong hydroxyl radical scavenging effects in the Fenton reaction system and nitric oxide suppressing effects in LPS-induced RAW 264.7 cells. Most tea flowers contain less caffeine, but comparable amounts of total catechins, compared to tea leaves and teas. The present study demonstrates that both tea flowers and tea leaves contain appreciable amounts of catechins and caffeine. It is likely that tea flowers might be useful for making alternative tea beverages.     

Uptake and distribution of manganese applied to leaves of Vicia faba (cv. Herzfreya) and Zea mays (cv. Regent) plants

Uptake and transport of Mn applied to leaves was studied in maize (Zea mays cv. Regent) and horse bean (Vicia faba cv. Herzfreya), two important crops in Egypt. Under controlled conditions in a growth chamber, maize and horse bean plants were grown in solution culture without Mn. After 20 days, Mn was applied to one older leaf by submerging part of the leaf blade into 0.1 mM MnSO₄ or MnEDTA solution for 48 h. At harvest (24 h later), plants were divided into fractions and Mn concentrations and contents determined. Plants without Mn application served as control. Only 1% of the Mn supply was taken up. Most of it remained at the application zone. However, part of the Mn moved out of the leaf of application and was preferentially transported to the shoot apex. This was indicated by a up to 2 times higher Mn concentration of the youngest leaf. When Mn was applied as MnEDTA, Mn uptake was lower but translocation enhanced compared to MnSO₄. There were no consistent differences between the plant species although mobility of Mn seemed to be higher in maize. Although the amounts of Mn taken up and translocated were low, the results suggest that in these plant species, leaf-applied Mn may contribute to the Mn nutrition of new growth.     

Anthocyanins from maize (Zea mays) and reed canarygrass (Phalaris arundinacea)

Flowers of maize, Zea mays, and reed canarygrass, Phalaris arundinacea, contain the same anthocyanins: cyanidin 3-glucoside, cyanidin 3-(6' '-malonylglucoside), cyanidin 3-(3' ',6' '-dimalonylglucoside), peonidin 3-glucoside, peonidin 3-(6' '-malonylglucoside), and peonidin 3-(dimalonylglucoside). The latter pigment has previously not been reported to occur in plants. Structure elucidations were primarily based on homo- and heteronuclear two-dimensional NMR and electrospray MS. During the isolation procedure using various mixtures of H(2)O, CF(3)CO(2)H, and CH(3)OH, and during storage in NMR solvent (CF(3)CO(2)D/CD(3)OD; 1:19, v/v) methyl esterification of the free acid function of the malonyl units of the pigments occurs. The acylated anthocyanins constitute more than 80% and 40% of the anthocyanins in P. arundinacea and Z. mays, respectively. Flowers and leaves of maize, Zea mays, contain the same anthocyanins in nearly equal relative proportions.     

Stabilization of carbon from maize in a sandy soil in a long-term experiment

The sequestration of carbon (C) in soil is not completely understood, and quantitative information about the amounts of organic carbon in the various fractions and their rates of turnover could improve understanding. We aimed (i) to quantify the amounts of C derived from maize at various depths in the soil in a long‐term field experiment with and without fertilization using ¹³C/¹²C analysis, (ii) to model changes in the organic C, and (iii) to compare measured and modelled pools of C. The organic C derived from the maize was measured in soil samples collected to a depth of 65 cm from four plots, two of which had been under continuous maize and two under continuous rye during long‐term field experiments with NPK and without fertilization. The fractionation procedures included particle‐size fractionation and extractions in water and in pyrophosphate solution. We used the Rothamsted Carbon Model to model the dynamics of the carbon from ¹³C data. The amounts of C derived from maize in the Ap horizon after 39 years of continuous maize cropping were 9.5% of the total organic C (where unfertilized) and 14.0% where NPK had been applied. Fertilization did not affect the residence time of carbon in the soil. The amounts of C derived from maize in water extracts were 21% of the total organic C (where unfertilized) and 22% where NPK had been applied. The extracts that were soluble in pyrophosphate and insoluble in acid were depleted in C from maize (the amounts were 5% and 7% of the total organic C, respectively). The results of the ¹³C natural abundance technique were used to model the dynamics of the organic C. Both the total organic C and the C derived from maize in the particle‐size fraction 0–63 μm agreed well with the total and maize‐derived sums of the model pools ‘inert organic matter’, ‘humified organic matter’ and ‘microbial biomass’. The model suggested that 64% (unfertilized) or 53% (NPK) of the organic C in the Ap horizon were inert. Only one of three published equations to determine the size of the inert pool agreed well with these model results.     

Aroma chemicals isolated and identified from leaves of Aloe arborescens Mill. Var. Natalensis Berger.

Extracts from leaves of aloe (Aloe arborescens Mill. var. natalensis Berger) were obtained using two methods: steam distillation under reduced pressure followed by dichloromethane extraction (DRP) and simultaneous purging and extraction (SPE). A total of 123 aroma chemicals were identified in the extracts obtained by both methods using gas chromatography and gas chromatography/mass spectrometry. There were 42 alcohols, 23 terpenoids, 21 aldehydes, 9 esters, 8 ketones, 6 acids, 5 phenols, and 9 miscellaneous compounds. The major aroma constituents of this extract by DRP were (Z)-3-hexenol (29.89%), (Z)-3-hexenal (18.86%), (E)-hexenal (7.31%), 4-methyl-3-pentenol (5.66%), and butanol (4.29%). The major aroma constituents of this extract by SPE were (E)-2-hexenal (45.46%), (Z)-3-hexenal (32.12%), hexanal (9.14%), (Z)-3-hexenol (1.60%), and 3-pentanone (1.41%). Terpenoids were also found as one of the major constituents. The fresh green note of aloe leaves is due to the presence of these C(6) alcohols and aldehydes as well as terpenoids.     

Antioxidant capacity and phenolic content in leaf extracts of tree spinach (Cnidoscolus spp.)

Total phenolic content and antioxidant capacity of two tree spinach species (Cnidoscolus chayamansa McVaugh and C. aconitifolius Miller.) were determined in raw and cooked leaf extracts. Antioxidant capacity was assessed by the oxygen radical absorbance capacity (ORAC) assay, and flavonoid glycoside composition was quantified by HPLC and identified by GC. Total phenolics and antioxidant capacity were higher in raw than in cooked leaf extracts. The ORAC values were strongly correlated with total phenolic content (r = 0.926) in all leaf extracts. The major flavonoids isolated from the leaf extracts were kaempferol-3-O-glycosides and quercetin-3-O-glycosides. C. aconitifolius leaves contained more varieties of the flavonoid glycosides than C. chayamansa. Cooking reduced antioxidant activity and phenolic content and resulted in losses of some kaempferol glycoside and quercetin glycoside residues in leaf extracts. The results of this study indicate that tree spinach leaves are a rich source of natural antioxidants for foods.     

Extraction and Characterization of Lipids from the Kernels,Leaves, and Stalks of Nine Grain Sorghum Parent Lines

The objectives of this study were to identify and quantify lipid classes extracted from the stalk, leaves, and the whole kernels of nine different sorghum parent lines. All extractions were performed with a Soxtec using n-hexane as the extraction solvent. Analytical methods including high-performance liquid chromatography (HPLC), gas chromatography (GC), and thin-layer chromatography (TLC) were used to identify and quantify the extracted lipid classes: triacylglycerides (TAG), diglycerides (DG), policosanols (PA), free sterols (FS), phytosterols (PS), free fatty acids (FFA), tocopherols (T), and fatty aldehydes. Overall, extracts from the leaves had higher amounts of soluble matter. Extracts from leaves contained more alcohols and sterols, whereas extracts from the stalks and the whole kernels contained the greatest amount of free fatty acids and fatty aldehydes, respectively. Trace amounts of tocopherol were detected by HPLC in all 27 samples. The lipid profiles of extracts from the whole kernels, leaves, and stalks from the same plant differed. The lipid profiles of extracts from each anatomical part of the sorghum plant differed between the nine parent lines.     

Genotypic differences in uptake and translocation of cadmium in bean and maize inbred lines

For better understanding of mechanisms responsible for genotypic differences in uptake and translocation of cadmium (Cd) in different plant species, two maize (Zea mays L.) inbred lines (B37 and F2) and a bean (Phaseolus vulgaris L.) cultivar (Saxa) were grown in a complete nutrient solution with additional 0.5 μM Cd and 250 μM buthionine sulfoximine (BSO), an inhibitor of PC synthesis, alone or in combination. The maize line B37 had a much higher Cd content in shoots (116.2 mg Cd kg^?1 dry wt.) than F2 (32.7 mg Cd kg^?1 dry wt.) and bean (1.83 in leaves, and 2.85 mg Cd kg^?1 dry wt. in stems), whereas in roots the Cd content was much higher in bean (602.6 mg Cd kg^?1 dry wt.) than in maize (427.1 mg Cd kg^?1 dry wt. in B37, and 428.2 mg Cd kg^?1 dry wt. in F2). Application of BSO markedly decreased Cd contents in roots of bean and maize lines, and also Cd contents in shoots and stem basis of both maize lines, while Cd contents in leaves, stems and stem basis of bean were not reduced by BSO. In root extracts (Tris-HCl buffer, pH 8.0) the proportion of Cd in the soluble fraction was much lower in bean (29.6%) than in the maize lines B37 (58.6%) and F2 (60.1%). Compared with the whole root tissue, Cd contents in the stele of the roots were much lower, especially in bean, and decreased by BSO in both maize lines, but not in bean. Gel-chromatography of root extracts strongly suggested that in the soluble fraction about 80% of the Cd was present as Cd-phytochelatin (PC) complexes in B37, whereas in F2 this Cd fraction accounted for about 50%, and in bean only for a few percent in the soluble fraction, Our results suggest that Cd-PC complexes constitute a mobile form in plants. The lower proportion of Cd in the soluble fraction as well as lower PC production in roots of bean compared to maize lines may be the main reasons for the very low Cd translocation from roots to shoots in bean plants.     

Antioxidant properties of various solvent extracts of total phenolic constituents from three different agroclimatic origins of drumstick tree (Moringa oleifera Lam.) leaves

Water, aqueous methanol, and aqueous ethanol extracts of freeze-dried leaves of Moringa oleifera Lam. from different agroclimatic regions were examined for radical scavenging capacities and antioxidant activities. All leaf extracts were capable of scavenging peroxyl and superoxyl radicals. Similar scavenging activities for different solvent extracts of each collection were found for the stable 1,1-diphenyl 2-picrylhydrazyl (DPPH(*)) radical. Among the three different moringa samples, both methanol and ethanol extracts of Indian origins showed the highest antioxidant activities, 65.1 and 66.8%, respectively, in the beta-carotene-linoleic acid system. Nonetheless, increasing concentration of all the extracts had significantly (P < 0.05) increased reducing power, which may in part be responsible for their antioxidant activity. The major bioactive compounds of phenolics were found to be flavonoid groups such as quercetin and kaempferol. On the basis of the results obtained, moringa leaves are found to be a potential source of natural antioxidants due to their marked antioxidant activity. This is the first report on the antioxidant properties of the extracts from freeze-dried moringa leaves. Overall, both methanol (80%) and ethanol (70%) were found to be the best solvents for the extraction of antioxidant compounds from moringa leaves.