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Identification of the main phenolic compounds in wood of Ceratonia siliqua by GC-MS.

The tannin composition of wood of Ceratonia siliqua (carob) was studied using GC-MS and classical chemical assays. Aqueous methanolic extracts of carob heartwood and sapwood were fractionated using organic solvents of increasing polarity, and GC-MS analyses were performed before and after hydrolysis. Prior to hydrolysis, gallic acid, catechin and its derivatives, methyl inositol and chalcone were determined as the major compounds found in the free form. Aqueous fractions of both wood types were hydrolysed with hydrochloric acid in methanol and extracted with organic solvents and water. These fractions were rich in methyl inositol, gallic acid, glucose and other monosaccharides. The results show that carob wood contains predominantly gallotannins and proanthocyanidins. The technique employed is shown to be a valuable tool and an alternative method to HPLC determination of hydrolysable tannin composition.

Fabaceae↗

Screening South Indian medicinal plants for antifungal activity against cutaneous pathogens.

In this study, twenty-eight South Indian medicinal plants were screened for their anti-fungal activity against six species of fungi (Trichophyton mentagrophytes, T. rubrum, T. soudanense, Candida albicans, Torulopsis glabrata, and C. krusei). Three plant species extracts, Celastrus paniculatus, Eriodendron anfractuosum and Ficus glomerata showed inhibitory activity. An aqueous extract of galls of Terminalia chebula showed inhibitory effects on three dermatophytes (Trichophyton spp.) and three yeasts (Candida spp.). Seeds extract of T. chebula inhibited only the growth of T. glabrata. An aqueous extract of T. chebula showed inhibitory effects higher than those measured in ethanol extracts. It is therefore suggested that tannins are plausible candidates for the anti-dermatophytic effects of T. chebula. Chebulinic acid, a known tannin of T. chebula was tested and found not inhibitory, thus a search for the active compound is needed.

Antifungal Agents↗

Phytochemicals from traditional medicinal plants used in the treatment of diarrhoea: modes of action and effects on intestinal function.

Medicinal plants have been used as traditional treatments for numerous human diseases for thousands of years. Diarrhoeal diseases continue to be a major cause of morbidity and mortality throughout the world and there is renewed interest in the discovery of novel compounds that can be used to fight these diseases. Numerous studies have validated the traditional use of antidiarrhoeal medicinal plants by investigating the biological activity of extracts of such plants, which have antispasmodic effects, delay intestinal transit, suppress gut motility, stimulate water adsorption or reduce electrolyte secretion. Of the numerous phytochemicals (such as alkaloids, tannins, flavonoids and terpenes) present in active extracts, tannins and flavonoids are thought to be responsible for antidiarrhoeal activity by increasing colonic water and electrolyte reabsorption. Others act by inhibiting intestinal motility. As some of the active ingredients are potentially toxic, there is a need to evaluate the safety of plant preparations. A few clinical trials have evaluated the safety and tolerability of traditional and herbal medicine preparations used to treat diarrhoea and generally indicate that minimal side effects are observed. However, with the increased popularity of plant-derived and herbal medicines in Western society, the benefits and potential dangers of these medicines must be considered.

Animals↗

Preservation of 2-D crystals of tubulin for electron crystallography.

Zinc-induced sheets of tubulin are two-dimensional crystalline polymers that constitute an ideal sample for high resolution studies of tubulin by electron crystallography. We show that these 2-dimensional tubulin crystals can be stabilized by taxol against low-temperature depolymerization and degradation with time, easing the way for the preparation of electron microscopy samples. The preservation of the crystals to high resolution has been tested with different embedding media. While glucose-embedded samples diffract poorly, samples embedded in tannin consistently diffract to a resolution of at least 3.5 A. Even better results are obtained by embedding with a combination of tannin and glucose, which improves the flatness of the crystals and allows the collection of isotropic high-resolution data from tilted specimens.

Animals↗

Improved methods for analysis and biological characterization of fiber.

Dietary fibers are not uniform, chemically or in their nutritive and biological properties, the only common ground being their resistance to mammalian digestive enzymes. The AOAC method for total fiber is subject to inferences from ash, protein, tannins and resistant starches. These interferences can be reduced by urea enzymatic dialysis. The measurement of soluble and insoluble fiber is nutritionally relevant, since physical properties greatly modify dietary effects of fiber. Insoluble fiber is conveniently measured as neutral-detergent fiber. This procedure has been improved by reducing the starch interference and the time of analysis. Physical and biological properties of dietary fiber can be measured by using relevant procedures for hydration capacity, metal ion exchange capacity and rate of fermentation. The lignin and tannin content modify the characteristics of dietary fiber.

Amylases↗

Putting the insect into the birch-insect interaction.

Leaf maturation in mountain birch (Betula pubescens ssp. czerepanovii) is characterized by rapid shifts in the types of dominant phenolics: from carbon-economic flavonoids aglycons in flushing leaves, via hydrolysable tannins and flavonoid glycosides, to carbon-rich proanthocyanidins (condensed tannins) in mature foliage. This shift accords with the suggested trade-offs between carbon allocation to plant defense and growth, but may also relate to the simultaneous decline in nutritive leaf traits, such as water, proteins and sugars, which potentially limit insect growth. To elucidate how birch leaf quality translates into insect growth, I introduce a simple model that takes into account defensive compounds but also acknowledges insect demand for nutritive compounds. The effects of defensive compounds on insect growth depend strongly on background variation in nutritive leaf traits: compensatory feeding on low nutritive diets increases the intake of defensive compounds, and the availability of growth-limiting nutritive compounds may modify the effects of defenses. The ratio of consumption to larval growth (both in dry mass) increases very rapidly with leaf maturation: from 2.9 to 9.8 over 2 weeks in June-July, and to 15 by August. High concentrations in mature birch leaves of "quantitative" defenses, such as proanthocyanidins (15-20% of dry mass), presumably prevent further consumption. If the same compounds had also protected half-grown leaves (which supported the same larval growth with only one third of the dry matter consumption of older leaves), the same intake of proanthocyanidins would have demanded improbably high concentrations (close to 50%) in young leaves. The model thus suggests an adaptive explanation for the high levels of "quantitative" defenses, such as proanthocyanidins, in low-nutritive but not in high-nutritive leaves because of the behavioral responses of insect feeding to leaf nutritive levels.

Adaptation, Physiological↗

Effects of nutrient variability on the genetic-based resistance of Eucalyptus globulus to a mammalian herbivore and on plant defensive chemistry.

Plant resistance to herbivores can be influenced not only by the independent effects of plant genotype and environmental variation, but by interactions between the two. The main aim of this study was to assess the effects of environmental variability (nutrient treatment) on the known genetic-based expression of resistance and defensive chemistry of Eucalyptus globulus to browsing by the generalist mammalian herbivore Trichosurus vulpecula. In a captive feeding trial, we measured intake of seedlings from one relatively resistant (Blue Gum Hill) and one relatively susceptible (St Helens) population of E. globulus grown under two nutrient treatments (no fertiliser, plus fertiliser). There was a significant genotypexfertiliser interaction effect on intake of E. globulus foliage by T. vulpecula, and the predicted genetic-based resistance of the two populations was expressed only for the non-fertilised treatment. Expression of resistance largely reflected the combined and inverse effects of nitrogen and condensed tannin concentrations. The expression of plant secondary metabolite concentration differed between compounds, but in all cases the effects of plant genotype and fertiliser treatment were independent. The formylated phloroglucinol compounds differed significantly between genotypes but not between fertiliser treatments. In contrast, the effect of plant genotype on the expression of condensed tannins was weak but they were significantly reduced by fertiliser. Essential oils were influenced by both plant genotype and fertiliser treatment and were significantly higher in the fertilised seedlings than in the non-fertilised seedlings. This study highlights interactive effects of plant genotype and environment in influencing the phenotypic expression of resistance in a eucalypt species to a mammalian browser. It also demonstrates that this interactive effect is the net result of independent effects of genotype and environment on plant chemistry and finally, that different groups of compounds within a plant can respond very differently to variation in environmental conditions.

Adaptation, Physiological↗

Induced resistance in the indeterminate growth of aspen (Populus tremuloides).

Studies of induction in trees have examined rapid induced resistance (RIR) or delayed induced resistance (DIR), but have not examined induction that occurs in leaves produced by indeterminately growing trees subsequent to, but in the same season as, damage. We refer to induction that occurs during this time period as intermediate-delayed induced resistance (IDIR). We assessed the influences of genetic and environmental factors, and their interactions, on temporal and spatial variation in induction and on tradeoffs between induced and constitutive levels of resistance in indeterminately growing saplings of aspen (Populus tremuloides). We utilized a common garden of 12 aspen genotypes experiencing two levels of defoliation and two levels of soil nutrients. We assessed concentrations of phenolic glycosides and condensed tannins in damaged leaf remnants collected 1 week after defoliation to examine rapid and local induction, and in undamaged leaves produced 8 weeks after defoliation to assess intermediate-delayed and systemic induction. In general, tannins showed RIR, while phenolic glycosides expressed IDIR. For both classes of allelochemicals, we found high estimates of broad-sense heritability and genetic variation in both induced and constitutive levels. Genetic variation may be maintained by both direct costs of allelochemicals and by costs of inducibility (phenotypic plasticity). Such costs may drive the tradeoff exhibited between induced and constitutive levels of phenolic glycosides. IDIR may be important in reducing total-season tissue loss by providing augmented resistance against late summer herbivores in trees that have experienced damage earlier in the season. Herbivore-resistant compensatory growth is especially beneficial to young trees growing in competitive environments.

Animals↗

Fine root chemistry and decomposition in model communities of north-temperate tree species show little response to elevated atmospheric CO2 and varying soil resource availability.

Rising atmospheric [CO2] has the potential to alter soil carbon (C) cycling by increasing the content of recalcitrant constituents in plant litter, thereby decreasing rates of decomposition. Because fine root turnover constitutes a large fraction of annual NPP, changes in fine root decomposition are especially important. These responses will likely be affected by soil resource availability and the life history characteristics of the dominant tree species. We evaluated the effects of elevated atmospheric [CO2] and soil resource availability on the production and chemistry, mycorrhizal colonization, and decomposition of fine roots in an early- and late-successional tree species that are economically and ecologically important in north temperate forests. Open-top chambers were used to expose young trembling aspen (Populus tremuloides) and sugar maple (Acer saccharum) trees to ambient (36 Pa) and elevated (56 Pa) atmospheric CO2. Soil resource availability was composed of two treatments that bracketed the range found in the Upper Lake States, USA. After 2.5 years of growth, sugar maple had greater fine root standing crop due to relatively greater allocation to fine roots (30% of total root biomass) relative to aspen (7% total root biomass). Relative to the low soil resources treatment, aspen fine root biomass increased 76% with increased soil resource availability, but only under elevated [CO2]. Sugar maple fine root biomass increased 26% with increased soil resource availability (relative to the low soil resources treatment), and showed little response to elevated [CO2]. Concentrations of N and soluble phenolics, and C/N ratio in roots were similar for the two species, but aspen had slightly higher lignin and lower condensed tannins contents compared to sugar maple. As predicted by source-sink models of carbon allocation, pooled constituents (C/N ratio, soluble phenolics) increased in response to increased relative carbon availability (elevated [CO2]/low soil resource availability), however, biosynthetically distinct compounds (lignin, starch, condensed tannins) did not always respond as predicted. We found that mycorrhizal colonization of fine roots was not strongly affected by atmospheric [CO2] or soil resource availability, as indicated by root ergosterol contents. Overall, absolute changes in root chemical composition in response to increases in C and soil resource availability were small and had no effect on soil fungal biomass or specific rates of fine root decomposition. We conclude that root contributions to soil carbon cycling will mainly be influenced by fine root production and turnover responses to rising atmospheric [CO2], rather than changes in substrate chemistry.

Atmosphere↗

Long-term experimental warming, shading and nutrient addition affect the concentration of phenolic compounds in arctic-alpine deciduous and evergreen dwarf shrubs.

Environmental changes are likely to alter the chemical composition of plant tissues, including content and concentrations of secondary compounds, and thereby affect the food sources of herbivores. After 10 years of experimental increase of temperature, nutrient levels and light attenuation in a sub-arctic, alpine ecosystem, we investigated the effects on carbon based secondary compounds (CBSC) and nitrogen in one dominant deciduous dwarf shrub, Salix herbacea x polaris and two dominant evergreen dwarf shrubs, Cassiope tetragona and Vaccinium vitis-idaea throughout one growing season. The main aims were to compare the seasonal course and treatment effects on CBSC among the species, life forms and leaf cohorts and to examine whether the responses in different CBSC were consistent across compounds. The changes in leaf chemistry both during the season and in response to the treatments were higher in S. herbacea x polaris than in the corresponding current year's leaf cohort of the evergreen C. tetragona. The changes were also much higher than in the 1-year-old leaves of the two evergreens probably due to differences in dilution and turnover of CBSC in growing and mature leaves paired with different rates of allocation. Most low molecular weight phenolics in the current year's leaves decreased in all treatments. Condensed tannins and the tannin-to-N ratio, however, either increased or decreased, and the strength and even direction of the responses varied among the species and leaf cohorts, supporting views of influential factors additional to resource-based or developmental controls, as e.g. species specific or genetic controls of CBSC. The results indicate that there is no common response to environmental changes across species and substances. However, the pronounced treatment responses imply that the quality of the herbivore forage is likely to be strongly affected in a changing arctic environment, although both the direction and strength of the responses will be different among plant species, tissue types and substances.

Arctic Regions↗

Do carbon-based defences reduce foliar damage? Habitat-related effects on tree seedling performance in a temperate rainforest of Chiloé Island, Chile.

Carbon-based secondary compounds (CBSCs), such as phenols or tannins, have been considered as one of the most important and general chemical barriers of woody plants against a diverse array of herbivores. Herbivory has been described as a critical factor affecting the growth and survival of newly established tree seedlings or juveniles then, the presence of secondary metabolites as defences against herbivores should be a primary strategy to reduce foliar damage. We examined whether light-induced changes in leaf phenolic chemistry affected insect herbivory on seedlings of two rainforest tree species, Drimys winteri (Winteraceae) and Gevuina avellana (Proteaceae). Seedlings of both species were planted under closed canopy and in a canopy gap within a large remnant forest patch. Half of the seedlings in each habitat were disinfected with a wide-spectrum systemic insecticide and the other half were used as controls. Seedling growth, survival, and foliar damage (estimated by an herbivory index) due to insect herbivores were monitored over a period of 16 months (December 2001-April 2003). The total leaf content of phenols and condensed tannins were assessed in seedlings from both habitats. As expected, access to light induced a greater production of CBSCs in seedlings of both tree species, but these compounds did not seem to play a significant defensive role, as seedlings grown in gaps suffered greater leaf damage than those planted in forest interior. In addition, in both habitats, seedlings without insecticide treatment suffered a greater foliar damage than those with insecticide, especially 16 months after the beginning of the experiment. Canopy openness and herbivory had positive and negative effects, respectively, on seedling growth and survival in both tree species. In conclusion, despite the higher levels of defence in tree-fall gap, the higher densities of herbivore override this and lead to higher damage levels.

Animals↗

Changes in resource concentration and defence during leaf development in a tough-leaved (Nothofagus moorei) and soft-leaved (Toona ciliata) species.

Developing leaves that are soft, with high concentrations of resources, can be particularly vulnerable to herbivore damage. Since a developing leaf cannot be very tough, given the constraints of cell expansion, the major form of protection is likely to be chemical defence. We investigated changes in concentration of herbivore resources (protein, carbohydrates and water) and putative defences (total phenolics, tannin activity, cyanogenic glycosides, alkaloids, cell wall, and leaf mechanics) across five leaf development stages of the soft-leaved Toona ciliata M. Roem. and the tough-leaved Nothofagus moorei (F. Muell.) Krasser. Chemical defences were predicted to be more highly developed in young than expanded leaves of both species, and to decline more in expanded leaves of N. moorei, which become tough and strong at maturity, than in the softer expanded leaves of T. ciliata. Resources and defences were dynamic within the developing leaves. Highest concentrations of protein were recorded in young leaves in both species, and highest levels of non-structural carbohydrate were recorded in young leaves of T. ciliata. Allocation to defence varied in both amount and type across leaf stages. In T. ciliata, there was an increase in chemical defence in expanded leaves (tannin activity, alkaloids). However, in N. moorei, increasing strength and toughness of developing leaves coincided with decreasing chemical defence, consistent with our hypothesis. For phenolics, this decrease was partly due to dilution by cell wall, but cyanogenic glycosides were present in young leaves and absent in fully mature leaves. These results are consistent with leaf toughness acting as an effective anti-herbivore defence, thereby reducing the need for investment in chemical defence.

Chlorophyll↗

Genotype and environment determine allocation to and costs of resistance in quaking aspen.

Although genetic variability and resource availability both influence plant chemical composition, little is known about how these factors interact to modulate costs of resistance, expressed as negative correlations between growth and defense. We evaluated genotype x environment effects on foliar chemistry and growth of quaking aspen (Populus tremuloides) by growing multiple aspen genotypes under variable conditions of light and soil nutrient availability in a common garden. Foliage was analyzed for levels of nitrogen, phenolic glycosides and condensed tannins. Bioassays of leaf quality were conducted with fourth-stadium gypsy moth (Lymantria dispar) larvae. Results revealed strong effects of plant genotype, light availability and nutrient availability; the importance of each factor depended upon compound type. For example, tannin concentrations differed little among genotypes and across nutrient regimes under low light conditions, but markedly so under high light conditions. Phenolic glycoside concentrations, in contrast, were largely determined by genotype. Variation in phenolic glycoside concentrations among genotypes was the most important factor affecting gypsy moth performance. Gypsy moth biomass and development time were negatively and positively correlated, respectively, with phenolic glycoside levels. Allocation to phenolic glycosides appeared to be costly in terms of growth, but only under resource-limiting conditions. Context-dependent trade-offs help to explain why costs of allocation to resistance are often difficult to demonstrate.

Animals↗

Tall herb herbivory resistance reflects historic exposure to leaf beetles in a boreal archipelago age-gradient.

In this paper, we introduce the coevolution-by-coexistence hypothesis which predicts that the strength of a coevolutionary adaptation will become increasingly apparent as long as the corresponding selection from an interacting counterpart continues. Hence, evolutionary interactions between plants and their herbivores can be studied by comparing discrete plant populations with known history of herbivore colonization. We studied populations of the host plant, Filipendula ulmaria (meadow sweet), on six islands, in a Bothnian archipelago subject to isostatic rebound, that represent a spatio-temporal gradient of coexistence with its two major herbivores, the specialist leaf beetles Galerucella tenella and Altica engstroemi. Regression analyses showed that a number of traits important for insect-plant interactions (leaf concentrations of individual phenolics and condensed tannins, plant height, G. tenella adult feeding and oviposition) were significantly correlated with island age. First, leaf concentrations of condensed tannins and individual phenolics were positively correlated with island age, suggesting that plant resistance increased after herbivore colonization and continued to increase in parallel to increasing time of past coexistence, while plant height showed a reverse negative correlation. Second, a multi-choice experiment with G. tenella showed that both oviposition and leaf consumption of the host plants were negatively correlated with island age. Third, larvae performed poorly on well-defended, older host populations and well on less-defended, younger populations. Thus, no parameter assessed in this study falsifies the coevolution-by-coexistence hypothesis. We conclude that spatio-temporal gradients present in rising archipelagos offer unique opportunities to address evolutionary interactions, but care has to be taken as abiotic (and other biotic) factors may interact in a complicated way.

Animals↗

Effects of elevated CO2 on foliar quality and herbivore damage in a scrub oak ecosystem.

Atmospheric CO2 concentrations have increased exponentially over the last century and continuing increases are expected to have significant effects on ecosystems. We investigated the interactions among atmospheric CO2, foliar quality, and herbivory within a scrub oak community at the Kennedy Space Center, Florida. Sixteen plots of open-top chambers were followed; eight of which were exposed to ambient levels of CO2 (350 ppm), and eight of which were exposed to elevated levels of CO2 (700 ppm). We focused on three oak species, Quercus geminata, Quercus myrtifolia, Quercus chapmanii, and one nitrogen fixing legume, Galactia elliottii. There were declines in overall nitrogen and increases in C:N ratios under elevated CO2. Total carbon, phenolics (condensed tannins, hydrolyzable tannins, total phenolics) and fiber (cellulose, hemicellulose, lignin) did not change under elevated CO2 across plant species. Plant species differed in their relative foliar chemistries over time, however, the only consistent differences were higher nitrogen concentrations and lower C:N ratios in the nitrogen fixer when compared to the oak species. Under elevated CO2, damage by herbivores decreased for four of the six insect groups investigated. The overall declines in both foliar quality and herbivory under elevated CO2 treatments suggest that damage to plants may decline as atmospheric CO2 levels continue to rise.

Atmosphere↗

Effects of elevated co2 and herbivore damage on litter quality in a scrub oak ecosystem.

Atmospheric CO(2) concentrations have increased dramatically over the last century and continuing increases are expected to have significant, though currently unpredictable, effects on ecosystems. One important process that may be affected by elevated CO(2) is leaf litter decomposition. We investigated the interactions among atmospheric CO(2), herbivory, and litter quality within a scrub oak community at the Kennedy Space Center, Florida. Leaf litter chemistry in 16 plots of open-top chambers was followed for 3 years; eight were exposed to ambient levels of CO(2), and eight were exposed to elevated levels of CO(2) (ambient + 350 ppmV). We focused on three dominant oak species, Quercus geminata, Quercus myrtifolia, and Quercus chapmanii. Condensed tannin concentrations in oak leaf litter were higher under elevated CO(2). Litter chemistry differed among all plant species except for condensed tannins. Phenolic concentrations were lower, whereas lignin concentrations and lignin/nitrogen ratios were higher in herbivore-damaged litter independent of CO(2) concentration. However, changes in litter chemistry from year to year were far larger than effects of CO(2) or insect damage, suggesting that these may have only minor effects on litter decomposition.

Animals↗

Resource availability and quality influence patterns of diet mixing by sheep.

In grazing systems, forage availability is a function of herbivore density, which can influence an animal's ability to be selective. In turn, the influence of food availability on selectivity has the potential to influence plant biodiversity. We hypothesized that the ability of herbivores to mix toxin-containing foods in their diets is a function of the availability of nontoxic foods and the nutritional characteristics of the toxin-containing foods. We evaluated this hypothesis in two trials simulating different diet qualities (high-quality foods in trial 1, low-quality foods in trial 2). Within each trial, the four treatment groups were offered with different amounts of nutritious, familiar foods-10, 30, 50, and 70% of ad libitum intake-but were offered with ad libitum access to toxin-containing foods. Each lamb was presented with five foods, including three toxin-containing unfamiliar foods (terpenes, tannins, and oxalates) and two nutritious familiar foods (alfalfa and barley). In trial 1, as the availability of nutritious familiar foods decreased, animals ate more of all three toxin-containing foods. As the availability of nutritious alternatives increased, the pattern of selection shifted from terpenes to tannins and oxalates. In trial 2, animals also ate more toxins as the availability of nutritious alternatives decreased, but their pattern of diet mixing changed. Low availability of nutritious alternatives resulted in the animals eating more oxalates. During preference tests when all five foods were offered ad libitum, animals fed with 10, 30, 50, and 70% of ad libitum intake from trial 1 ate 49, 47, 41, and 38% of the three toxin-containing foods, respectively. The lower diet quality in trial 2 affected intake of the toxin-containing foods such that animals fed with 10, 30, 50, and 70% of ad libitum intake ate 37, 36, 29, and 27%, respectively, of the three toxin-containing foods. Thus, the quality of toxin-containing foods and the availability of nutritious alternatives interacted to modify the pattern of diet mixing by lambs.

Animals↗

An unusual case of black teeth.

Loss of enamel and a deep black stain of the teeth in a 40-year-old diabetic patient are strongly suggested to be caused by the daily consumption of a cheap white wine and, possibly, by the chewing of cayenne. The wine proved to be rather acid, thereby promoting abrasion as a result of gnashing, and to contain a high concentration of tannin. The exact role of the tannins is described.

Adult↗