Novel oligosaccharides isolated from Fusarium oxysporum L. rapidly induce PAL activity in Rubus cells.
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Activation of the phenolic pathway is known to be part of a defense response against cell wall-derived elicitors from pathogens. Many examples of a defense response by increasing the synthesis of phenolic compound against the elicitor were demonstrated in the past, but the elicitor structure has so far been poorly characterized. Our results indicate that a disaccharide fraction containing the following structure: alpha-D-mannopyranosyl (1-->2)alpha/beta-D-glucopyranosyl and alpha-D-mannopyranosyl (1-->x) inositol, isolated from Fusarium oxysporum L., promotes rapid and transient phenylalanine ammonia lyase activity in Rubus fructicosus cells at nanomolar concentration. The disaccharides were isolated by size-exclusion chromatography directly from extracts obtained by alkaline treatment of F. oxysporum mycelium. Their structure was determined by 500-MHz-1H-NMR spectroscopy combined with methylation analysis and fast atom bombardment mass spectrometry. (+info)
Adhesion of human neutrophils to fibronectin is inhibited by comaruman, pectin of marsh cinquefoil Comarum palustre L., and by its fragments.
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Earlier, we detected antiinflammatory action of comaruman, pectin of the marsh cinquefoil Comarum palustre L. This effect can be explained by new data concerning inhibition of adhesion of human neutrophils to fibronectin by comaruman and its fragments. The galacturonan backbone fragment of molecular mass >10 kD appears to be the active region of the comaruman macromolecule. Comaruman CP (50-200 microg/ml) was found to decrease adhesion of neutrophils stimulated by phorbol 12-myristate 13-acetate (PMA, 1.625 microM) and by dithiothreitol (DTT, 0.5 mM). The fragments of comaruman CP-H (100 kD), CP-H1 (10-50 kD), and CP-H2 (100 kD) obtained by acidic hydrolysis and representing regions of linear polygalacturonan are shown to inhibit neutrophil adhesion more than the crude pectin. A fragment CP-E (<10 kD) obtained using pectinolysis and representing a branched region of the comaruman macromolecule failed to influence cell adhesion. The parent comaruman CP as well as fragments of its polygalacturonan backbone diminish PMA-initiated generation of oxygen radicals in neutrophils. (+info)
Detection and quantification of ligands involved in nickel detoxification in a herbaceous Ni hyperaccumulator Stackhousia tryonii Bailey.
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Field-collected, young plants of Ni hyperaccumulator Stackhousia tryonii, grown in a glasshouse for 20 weeks, were exposed to low- (available Ni concentration in the native serpentine soil, i.e. 60 microg g(-1) dry soil) and high- (external application of 1000 ppm) Ni concentrations in the substrate. Nickel concentration in the freeze-dried leaf tissues increased from 3700 microg g(-1) to 13 700 microg g(-1) with soil Ni supplementation, of which >60% was extracted with dilute acid (0.025 M HCl). Nickel supplementation also elicited a 575%, 211%, and 37% increase in the final concentrations of oxalic, citric, and malic acids, respectively, in leaf tissues. Malic acid was the dominant organic acid, followed by citric and oxalic acids. The molar ratio of Ni to malic acid was 1.0, consistent with a role for malate as a ligand for Ni in hyperaccumulating plants, supporting detoxification/transport and storage of this heavy metal in S. tryonii. The total amino acid concentrations in the xylem sap did not change with Ni supplementation (21.7+/-3.7 mM and 17.9+/-5 mM, respectively, for low- and high-nickel-treated plants). Glutamine was the major amino acid in both the low- and high-Ni-treated plants. The concentration of glutamine decreased by >60%, with a corresponding increase in alanine, aspartic acid, and glutamic acid, on exposure to high Ni. A role of amino acids in Ni complexation and transport in S. tryonii is not immediately apparent. (+info)
Expressed sequence tags (ESTs) and simple sequence repeat (SSR) markers from octoploid strawberry (Fragaria x ananassa).
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BACKGROUND: Cultivated strawberry (Fragaria x ananassa) represents one of the most valued fruit crops in the United States. Despite its economic importance, the octoploid genome presents a formidable barrier to efficient study of genome structure and molecular mechanisms that underlie agriculturally-relevant traits. Many potentially fruitful research avenues, especially large-scale gene expression surveys and development of molecular genetic markers have been limited by a lack of sequence information in public databases. As a first step to remedy this discrepancy a cDNA library has been developed from salicylate-treated, whole-plant tissues and over 1800 expressed sequence tags (EST's) have been sequenced and analyzed. RESULTS: A putative unigene set of 1304 sequences--133 contigs and 1171 singlets--has been developed, and the transcripts have been functionally annotated. Homology searches indicate that 89.5% of sequences share significant similarity to known/putative proteins or Rosaceae ESTs. The ESTs have been functionally characterized and genes relevant to specific physiological processes of economic importance have been identified. A set of tools useful for SSR development and mapping is presented. CONCLUSION: Sequences derived from this effort may be used to speed gene discovery efforts in Fragaria and the Rosaceae in general and also open avenues of comparative mapping. This report represents a first step in expanding molecular-genetic analyses in strawberry and demonstrates how computational tools can be used to optimally mine a large body of useful information from a relatively small data set. (+info)
Do xylem fibers affect vessel cavitation resistance?
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Possible mechanical and hydraulic costs to increased cavitation resistance were examined among six co-occurring species of chaparral shrubs in southern California. We measured cavitation resistance (xylem pressure at 50% loss of hydraulic conductivity), seasonal low pressure potential (P(min)), xylem conductive efficiency (specific conductivity), mechanical strength of stems (modulus of elasticity and modulus of rupture), and xylem density. At the cellular level, we measured vessel and fiber wall thickness and lumen diameter, transverse fiber wall and total lumen area, and estimated vessel implosion resistance using (t/b)(h)(2), where t is the thickness of adjoining vessel walls and b is the vessel lumen diameter. Increased cavitation resistance was correlated with increased mechanical strength (r(2) = 0.74 and 0.76 for modulus of elasticity and modulus of rupture, respectively), xylem density (r(2) = 0.88), and P(min) (r(2) = 0.96). In contrast, cavitation resistance and P(min) were not correlated with decreased specific conductivity, suggesting no tradeoff between these traits. At the cellular level, increased cavitation resistance was correlated with increased (t/b)(h)(2) (r(2) = 0.95), increased transverse fiber wall area (r(2) = 0.89), and decreased fiber lumen area (r(2) = 0.76). To our knowledge, the correlation between cavitation resistance and fiber wall area has not been shown previously and suggests a mechanical role for fibers in cavitation resistance. Fiber efficacy in prevention of vessel implosion, defined as inward bending or collapse of vessels, is discussed. (+info)
Phosphoenolpyruvate carboxykinase and its potential role in the catabolism of organic acids in the flesh of soft fruit during ripening.
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Previous studies of grapes and tomatoes have shown that the abundance of phosphoenolpyruvate carboxykinase (PEPCK) increases in their flesh at the start of ripening, and that this coincides with a decrease in its citrate and/or malate content. Thus, PEPCK might function in the catabolism of organic acid anions during the ripening of these fruits. In the present study, the abundance of PEPCK was determined in the flesh of blueberries, raspberries, red currants, and strawberries at different stages of their development. In addition, changes in the amounts of citrate, malate, soluble sugars, isocitrate lyase, NADP-malic enzyme, phosphoenolpyruvate carboxylase, and pyruvate, orthophosphate dikinase in the flesh were determined. PEPCK was not detected in strawberry flesh, in which there was no dissimilation of malate or citrate. In the flesh of the other fruits, the abundance of PEPCK increased during ripening to an amount that was similar to that in grapes and tomatoes. In the flesh of blueberries and red currants, PEPCK was most abundant when there was dissimilation of malate. In the flesh of raspberries, PEPCK was most abundant when there was dissimilation of malate and citrate. These results are consistent with PEPCK playing a role in the dissimilation of citrate and/or malate in the flesh of these fruits during ripening. However, PEPCK was also present in the flesh of blueberries, raspberries, and red currants when there was no dissimilation of malate or citrate, and this raises the possibility that PEPCK might have additional functions. Dissection of blueberries provided evidence that both PEPCK and phosphoenolpyruvate carboxylase were present in the same cells, and possible functions for this are discussed. (+info)
Anthocyanins exist in the circulation primarily as metabolites in adult men.
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Anthocyanins are reported to have many "health promoting" properties; however, despite numerous reports of their bioactivities, their absorption and metabolism in humans are poorly understood. The objective of this research was to detail the pharmacokinetic parameters of anthocyanins after the administration of a 721-mg oral dose of cyanidin 3-glycosides from chokeberry extract to human subjects. Solid-phase extraction, preparative-HPLC, preparative-TLC, HPLC-diode array detection, HPLC-MS, and NMR were utilized to isolate, identity, and quantify anthocyanins in 0- to 7-h (0, 1, 2, 3, 4, 5, 6, 7 h) serum and 0- to 24-h urine samples (total individual urine voids over 24 h). The cumulative concentration of total anthocyanins (parent and metabolites) detected in the serum (0-7 h) was 376.65 +/- 16.20 (nmol x h)/L (area under the concentration time curve), reaching a maximum concentration (C(max) = 96.08 +/- 6.04 nmol/L) within 2.8 h. The parent anthocyanins represented only 32.0% [120.63 +/- 2.85 (nmol x h)/L] of the total anthocyanins detected with 68.0% [256.02 +/- 5.23 (nmol x h) identified as conjugated metabolites. Additionally, the total urinary excretion of anthocyanins over 24 h was 1071.54 +/- 375.46 microg, reaching a maximal rate of excretion (R(max) = 202.74 +/- 85.06 microg/h) at 3.72 +/- 0.83 h. Parallel to the serum data, only 32.5% (347.85 +/- 60.61 microg) of the anthocyanins excreted in the urine (total 24 h) were the parent compounds with 67.5% (723.69 +/- 92.59 microg) occurring as conjugated metabolites. The metabolites were identified as glucuronidated and methylated derivatives of the parent cyanidin 3-glycosides. The above results indicate that cyanidin 3-glycosides are rapidly absorbed and metabolized extensively following a moderate-to-high oral dose in humans. (+info)
Effects of elevated atmospheric CO2 on soil microbial biomass, activity, and diversity in a chaparral ecosystem.
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This study reports the effects of long-term elevated atmospheric CO2 on root production and microbial activity, biomass, and diversity in a chaparral ecosystem in southern California. The free air CO2 enrichment (FACE) ring was located in a stand dominated by the woody shrub Adenostoma fasciculatum. Between 1995 and 2003, the FACE ring maintained an average daytime atmospheric CO2 concentration of 550 ppm. During the last two years of operation, observations were made on soil cores collected from the FACE ring and adjacent areas of chaparral with ambient CO2 levels. Root biomass roughly doubled in the FACE plot. Microbial biomass and activity were related to soil organic matter (OM) content, and so analysis of covariance was used to detect CO2 effects while controlling for variation across the landscape. Extracellular enzymatic activity (cellulase and amylase) and microbial biomass C (chloroform fumigation-extraction) increased more rapidly with OM in the FACE plot than in controls, but glucose substrate-induced respiration (SIR) rates did not. The metabolic quotient (field respiration over potential respiration) was significantly higher in FACE samples, possibly indicating that microbial respiration was less C limited under high CO2. The treatments also differed in the ratio of SIR to microbial biomass C, indicating a metabolic difference between the microbial communities. Bacterial diversity, described by 16S rRNA clone libraries, was unaffected by the CO2 treatment, but fungal biomass was stimulated. Furthermore, fungal biomass was correlated with cellulase and amylase activities, indicating that fungi were responsible for the stimulation of enzymatic activity in the FACE treatment. (+info)