Tree resistance to Lymantria dispar caterpillars: importance and limitations of foliar tannin composition

The ability of foliar tannins to increase plant resistance to herbivores is potentially determined by the composition of the tannins; hydrolyzable tannins are much more active as prooxidants in the guts of caterpillars than are condensed tannins. By manipulating the tannin compositions of two contrasting tree species, this work examined: (1) whether increased levels of hydrolyzable tannins increase the resistance of red oak (Quercus rubra L.), a tree with low resistance that produces mainly condensed tannins, and (2) whether increased levels of condensed tannins decrease the resistance of sugar maple (Acer saccharum Marsh.), a tree with relatively high resistance that produces high levels of hydrolyzable tannins. As expected, when Lymantria dispar L. caterpillars ingested oak leaves coated with hydrolyzable tannins, levels of hydrolyzable tannin oxidation increased in their midgut contents. However, increased tannin oxidation had no significant impact on oxidative stress in the surrounding midgut tissues. Although growth efficiencies were decreased by hydrolyzable tannins, growth rates remained unchanged, suggesting that additional hydrolyzable tannins are not sufficient to increase the resistance of oak. In larvae on condensed tannin-coated maple, no antioxidant effects were observed in the midgut, and levels of tannin oxidation remained high. Consequently, neither oxidative stress in midgut tissues nor larval performance were significantly affected by high levels of condensed tannins. Post hoc comparisons of physiological mechanisms related to tree resistance revealed that maple produced not only higher levels of oxidative stress in the midgut lumen and midgut tissues of L. dispar, but also decreased protein utilization efficiency compared with oak. Our results suggest that high levels of hydrolyzable tannins are important for producing oxidative stress, but increased tree resistance to caterpillars may require additional factors, such as those that produce nutritional stress.     

Feeding on poplar leaves by caterpillars potentiates foliar peroxidase action in their guts and increases plant resistance

Peroxidases (PODs) are believed to act as induced and constitutive defenses in plants against leaf-feeding insects. However, little work has examined the mode of action of PODs against insects. Putative mechanisms include the production of potentially antinutritive and/or toxic semiquinone free radicals and quinones (from the oxidation of phenolics), as well as increased leaf toughness. In this study, transgenic hybrid poplar saplings (Populus tremula × Populus alba) overexpressing horseradish peroxidase (HRP) were produced to examine the impact of elevated HRP levels on the performance and gut biochemistry of Lymantria dispar caterpillars. HRP-overexpressing poplars were more resistant to L. dispar than wild-type (WT) poplars when the level of a phenolic substrate of HRP (chlorogenic acid) was increased, but only when leaves had prior feeding damage. Damaged (induced) leaves produced increased amounts of hydrogen peroxide, which was used by HRP to increase the production of semiquinone radicals in the midguts of larvae. The decreased growth rates of larvae that fed on induced HRP-overexpressing poplars resulted from post-ingestive mechanisms, consistent with the action of HRP in their midguts. The toughness of HRP-overexpressing leaves was not significantly greater than that of WT leaves, whether or not they were induced. When leaves were coated with ellagitannins, induced HRP leaves also produced elevated levels of semiquinone radicals in the midgut. Decreased larval performance on induced HRP leaves in this case was due to post-ingestive mechanisms as well as decreased consumption. The results of this study provide the first demonstration that a POD is able to oxidize phenolics within an insect herbivore’s gut, and further clarifies the chemical conditions that must be present for PODs to function as antiherbivore defenses.     

Tannins in plant-herbivore interactions

Tannins are the most abundant secondary metabolites made by plants, commonly ranging from 5% to 10% dry weight of tree leaves. Tannins can defend leaves against insect herbivores by deterrence and/or toxicity. Contrary to early theories, tannins have no effect on protein digestion in insect herbivores. By contrast, in vertebrate herbivores tannins can decrease protein digestion. Tannins are especially prone to oxidize in insects with high pH guts, forming semiquinone radicals and quinones, as well as other reactive oxygen species. Tannin toxicity in insects is thought to result from the production of high levels of reactive oxygen species. Tannin structure has an important effect on biochemical activity. Ellagitannins oxidize much more readily than do gallotannins, which are more oxidatively active than most condensed tannins. The ability of insects to tolerate ingested tannins comes from a variety of biochemical and physical defenses in their guts, including surfactants, high pH, antioxidants, and a protective peritrophic envelope that lines the midgut. Most work on the ecological roles of tannins has been correlative, e.g., searching for negative associations between tannins and insect performance. A greater emphasis on manipulative experiments that control tannin levels is required to make further progress on the defensive functions of tannins. Recent advances in the use of molecular methods has permitted the production of tannin-overproducing transgenic plants and a better understanding of tannin biosynthetic pathways. Many research areas remain in need of further work, including the effects of different tannin types on different types of insects (e.g., caterpillars, grasshoppers, sap-sucking insects).     

Semiquinone and ascorbyl radicals in the gut fluids of caterpillars measured with EPR spectrometry

The biological activity of phenolic compounds ingested by caterpillars is commonly believed to result from their oxidation, although the products of oxidation have been well-characterized in only a few cases. The initial oxidation products of phenols (semiquinone or phenoxyl radicals) can be measured with electron paramagnetic resonance (EPR) spectrometry. In this study semiquinone radicals formed from tannic acid and gallic acid in the gut fluids of two species of caterpillars were measured. In Orgyia leucostigma, in which ingested phenols are not oxidized, semiquinone radicals were absent or at very low intensities. By contrast, in Malacosoma disstria, in which ingested phenols are oxidized, high semiquinone radical intensities were measured. In the absence of detectable levels of semiquinone radicals, ascorbyl radicals were detected in the EPR spectra instead. High molar ratios of ascorbate to phenols in an artificial diet produced ascorbyl radicals in the midgut fluids of both species, while diets containing low molar ratios produced semiquinone radicals. Similar results were obtained in M. disstria fed the leaves of red oak or sugar maple. The results of this study provide further evidence that ascorbate is an essential antioxidant that prevents the oxidation of phenols in the gut fluids of caterpillars, and demonstrate that EPR spectrometry is a valuable method for determining the degree of oxidative activation of phenols ingested by herbivorous insects.     

Fenton-type reactions and iron concentrations in the midgut fluids of tree-feeding caterpillars

Peroxides are formed in the midgut fluids of caterpillars when ingested tannins and other phenolic compounds oxidize. If these peroxides broke down in the presence of redox-active metal ions, they would form damaging free radicals (Fenton-type reactions). Elemental iron is present in relatively large amounts in leaves and artificial diets, but little is known about its concentration and redox state in midgut fluids, or the extent of Fenton-type reactions in these conditions. This study compared the levels of hydroxyl radicals and iron in the midgut fluids of two species of caterpillars: Orgyia leucostigma, in which phenol oxidation is limited, and Malacosoma disstria, in which phenol oxidation is more extensive. We tested two hypotheses: (1) higher levels of hydroxyl radicals are formed in M. disstria (consistent with the higher concentrations of hydrogen peroxide in this species), and (2) lower concentrations of iron are present in O. leucostigma (providing greater protection of its midgut fluids from oxidative damage). Hydroxyl radical levels increased greatly in M. disstria, but not in O. leucostigma, when they consumed a tannin-containing diet, supporting the first hypothesis. Protein oxidation was also significantly increased in the midgut fluids of M. disstria that ingested tannic acid, consistent with hydroxyl radical damage. Contrary to the second hypothesis, similar concentrations of iron (70 microM) remained in solution or suspension in both species of caterpillars on an artificial diet. Over 90% of this iron appeared to be in the reduced (catalytically active) state in both species. We conclude that tree-feeding caterpillars protect their midgut fluids from oxidative damage caused by Fenton-type reactions by limiting the formation of peroxides, rather than by limiting the availability of reduced iron.     

Gut pH, redox conditions and oxygen levels in an aquatic caterpillar: potential effects on the fate of ingested tannins

Larvae of the freshwater moth Acentria ephemerella (Pyralidae, Lepidoptera) can fully develop on Myriophyllum spicatum, a submerged macrophyte containing 7-10% of its dry mass as tannins. We investigated the physicochemical gut parameters of larvae fed with M. spicatum or Potamogeton perfoliatus, a food plant lacking tannins, and the chemical fate of ingested polyphenols. Microelectrode studies revealed that larval midguts were slightly alkaline (pH 8) and had a positive redox potential. Whole guts were oxygen sinks owing to the oxygen demand of the gut contents. Oxygen penetrated the midgut up to 100 microm, but the centres of the foregut and midgut were always anoxic. The physicochemical parameters of the guts did not change with the food plant. The major tannin from M. spicatum, tellimagrandin II, was significantly depleted in the midgut and was not detected in faeces. In vitro studies indicated that tellimagrandin II is rapidly depleted mainly through oxidation, and hydrolysis might also occur. Our findings for A. ephemerella are compared with those for terrestrial Lepidoptera, and possible mechanisms for adaptations to tannin-rich food plants are discussed.     

Comparison of midgut bacterial diversity in tropical caterpillars (Lepidoptera: Saturniidae) fed on different diets

As primary consumers of foliage, caterpillars play essential roles in shaping the trophic structure of tropical forests. The caterpillar midgut is specialized in plant tissue processing; its pH is exceptionally alkaline and contains high concentrations of toxic compounds derived from the ingested plant material (secondary compounds or allelochemicals) and from the insect itself. The midgut, therefore, represents an extreme environment for microbial life. Isolates from different bacterial taxa have been recovered from caterpillar midguts, but little is known about the impact of these microorganisms on caterpillar biology. Our long-term goals are to identify midgut symbionts and to investigate their functions. As a first step, different diet formulations were evaluated for rearing two species of tropical saturniid caterpillars. Using the polymerase chain reaction (PCR) with primers hybridizing broadly to sequences from the bacterial domain, 16S rRNA gene libraries were constructed with midgut DNA extracted from caterpillars reared on different diets. Amplified rDNA restriction analysis indicated that bacterial sequences recovered from the midguts of caterpillars fed on foliage were more diverse than those from caterpillars fed on artificial diet. Sequences related to Methylobacterium sp., Bradyrhizobium sp., and Propionibacterium sp. were detected in all caterpillar libraries regardless of diet, but were not detected in a library constructed from the diet itself. Furthermore, libraries constructed with DNA recovered from surface-sterilized eggs indicated potential for vertical transmission of midgut symbionts. Taken together, these results suggest that microorganisms associated with the tropical caterpillar midgut may engage in symbiotic interactions with these ecologically important insects.     

Condensed tannins and sugars in the diet of chimpanzees (Pan troglodytes schweinfurthii) in the Budongo Forest, Uganda

Fruits, leaves and bark forming part of the diet of chimpanzees were collected and it was noted whether samples were of a kind being eaten or not eaten. Samples were dried and analysed for condensed tannin content and for three sugars, glucose, sucrose and fructose. It was found that chimpanzees did not select foods according to the level of tannins but did so according to the levels of sugars, preferring the higher levels. Fig seeds contained higher tannin levels than fig pulp, and the chimpanzees made oral boli (“wadges”) of fig seeds which they spat out. Two fig species were compared: the one with lower tannin and higher sugar content was preferred. The bark of one tree species often eaten contained high levels of tannins but also contained sugars. Young leaves with lower tannin levels were preferred to mature leaves with higher levels. Chimpanzees appear to be able to tolerate higher tannin levels than three monkey species in this forest, and considerably higher levels than marmosets (Callitrichidae). Received: 20 October 1997 / Accepted: 1 March 1998     

Oligomeric hydrolyzable tannins from Monochaetum multiflorum

Four hydrolyzable tannins, nobotanins Q, R, S, and T, were isolated from the aqueous acetone extract of the dried leaves of Monochaetum multiflorum (Melastomataceae), a plant indigenous to Colombia. Their dimeric and tetrameric structures were elucidated by spectral and chemical methods. Eight known hydrolyzable tannin monomers and eight ellagitannin oligomers characteristic of melastomataceous plants were also characterized as tannin constituents of the plant.     

Induced sink strength as a prerequisite for induced tannin biosynthesis in developing leaves of Populus

Induced defenses occur predominately in young, developing plant tissues that rely upon carbohydrate import to support their growth and development. To test the hypothesis that the induced production of carbon-based defenses is dependent upon photoassimilate import, we examined the response of developing leaves of hybrid poplar (Populus deltoides 2 P. nigra) saplings to wounding by gypsy moth caterpillars (Lymantria dispar L.) and exogenous jasmonic acid (JA). Growth rates, condensed tannin contents and acid invertase activities were measured for individual leaves and the translocation of ¹³C-labeled resources between orthostichous source-sink pairs was quantified. Results showed a substantial increase in the activity of cell wall invertase in sink leaves wounded by gypsy moth caterpillars and treated with JA. JA-induced sink leaves also imported 3-4 times as much ¹³C-labeled carbon from orthostichous source leaves relative to controls and allocated a significant portion of this imported ¹³C to condensed tannin biosynthesis. Reduced carbohydrate flow to these leaves, caused by source leaf removal, resulted in reduced condensed tannin levels and the emergence of a growth-defense tradeoff. These results indicate that (1) induced sink strength is elicited by insect wounding and JA application in hybrid poplar foliage, (2) imported resources are allocated to the production of carbon-based defenses, and (3) the level of induced defense in leaves can be constrained by the ability of leaves to import carbohydrates from source tissues. Together, these results suggest that within-canopy variations in induced resistance may arise in part because of uneven distribution of resources to induced foliage.     

Effects of tree age and stand thinning related variations in balsam fir secondary compounds on spruce budworm Choristoneura fumiferana development,growth and food utilization

• 1 The effect of tannins and monoterpenes on the development, mortality and food utilization of spruce budworm Choristoneura fumiferana (Clem.) (Lepidoptera: Tortricidae) was investigated under laboratory conditions using an artificial diet. Tannins were extracted from balsam fir foliage of thinned and unthinned stands to reproduce stand thinning related variations in tannins. A mixture of synthetic monoterpenes was utilized to simulate the concentration found in young and old balsam fir trees.
• 2 Longer development time and lower pupal weight were observed for insects fed on diets with a lower nitrogen concentration and a higher tannin concentration (unthinned treatment). Tannins induced higher insect mortality at a low nitrogen concentration compared with the diet with a higher nitrogen concentration.
• 3 Approximate digestibility was higher for larvae fed on diets with high concentrations of nitrogen at both low and high concentrations of tannins. Efficiency of conversion of digested food (ECD) decreased with an increase in tannin concentration. Tannins reduced both the relative consumption and growth rate (RCR and RGR).
• 4 Monoterpenes increased spruce budworm mortality and this mortality reached almost 50% under concentrations of monoterpene typical of the young trees compared with 20% under monoterpene concentrations found in old trees.
• 5 A higher digestibility was observed for larvae fed on diet with a higher concentration of monoterpenes, whereas efficiency of conversion of ingested food (ECI), ECD, RCR, and RGR decreased with an increase in monoterpenes in the diet.
• 6 The results obtained in the present study are consistent with the defensive role of secondary compounds such as tannins and monoterpenes in the spruce budworm–balsam fir system.

     

Physiological factors affecting the rapid decrease in protein assimilation efficiency by a caterpillar on newly‐mature tree leaves

Lymantria dispar L. caterpillars have a decreased ability to assimilate protein from mature leaves of red oak (Quercus rubra) compared with young, expanding leaves. The present study determines whether the drop in protein assimilation efficiency (PAE) occurs during the rapid phase of leaf maturation. Several mechanisms that might account for decreased PAE are also examined: mature leaf tissues could resist being chewed efficiently, protein in mature leaf tissues could become difficult to extract, and other nutrients in mature leaves might become growth limiting. The entire seasonal decrease in PAE occurs rapidly (in less than 2 weeks), when the leaves finished expanding. The maturation process is characterized by increased levels of fibre and decreased levels of water but no significant changes in the levels of protein or carbohydrates. Despite increased fibre in mature leaves, they are not chewed into larger food particles than are immature leaves. Carbohydrate assimilation efficiencies remain high on mature leaves, and signs of limiting water levels in larvae of L. dispar on mature leaves are not observed. The most important finding in the present study is the decreased extractability of protein in food particles from mature leaves, which plays a major role in explaining the rapid decrease in PAE. It is hypothesized that structural changes in cell walls during the rapid process of leaf maturation decrease protein extractability, which, in turn, greatly decreases the nutritional quality of mature oak leaves for caterpillars. The results of the present study therefore suggest a general mechanism to help explain the widely documented decrease in the nutritional quality of the mature leaves of many tree species for herbivorous insects.     

Stem galls affect oak foliage with potential consequences for herbivory

Abstract.   1. On two dates, foliar characteristics of pin oak, Quercus palustris , infested with stem galls caused by the horned oak gall, Callirhytis cornigera , were investigated, and the consequences for subsequent herbivory assessed.
2. Second-instar caterpillars of the gypsy moth, Lymantria dispar , preferred foliage from ungalled trees.
3. Ungalled trees broke bud earlier than their galled counterparts.
4. Galled trees produced denser leaves with higher nitrogen and tannin concentrations, but foliar carbohydrates did not differ among galled and ungalled trees.
5. Concentrations of foliar carbohydrates in both galled and ungalled trees increased uniformly between the two assay dates. Nitrogen concentrations were greater in leaves from galled trees, and decreased uniformly in galled and ungalled trees over time. Foliar tannins were also greater in foliage from galled trees early in the season; however, foliar tannins declined seasonally in galled tissue so that by the second assay date there was no difference in tannin concentrations between galled and ungalled foliage.
6. In spite of differences in foliar characteristics, performance of older, fourth instar gypsy moth caterpillars did not differ between galled and ungalled trees.     

Importance of protein quality versus quantity in alternative host plants for a leaf-feeding insect

The nutritional value of alternative host plants for leaf-feeding insects such as caterpillars is commonly measured in terms of protein quantity. However, nutritional value might also depend on the quality of the foliar protein [i.e., the composition of essential amino acids (EAAs)]. A lack of comparative work on the EAA compositions of herbivores and their host plants has hampered the testing of this hypothesis. We tested the “protein quality hypothesis” using the tree-feeding caterpillars of Lymantria dispar (gypsy moth) and two taxonomically unrelated host plants, red oak (Quercus rubra) and sugar maple (Acer saccharum). Because L. dispar has higher fitness on oak than on maple, support for the hypothesis would be found if protein were of higher quality from oak than from maple. The whole-body EAA composition of L. dispar larvae was measured to estimate its optimum dietary protein composition, which was compared with the EAA compositions of oak and maple leaves. Contrary to the protein quality hypothesis, the EAA compositions of oak and maple were not significantly different in the spring. The growth-limiting EAAs in both tree species were histidine and methionine. Similar results were observed in the summer, with the exception that the histidine composition of oak was between 10 and 15 % greater than in maple leaves. The two main factors that affected the nutritional value of protein from the tree species were the quantities of EAAs, which were consistently higher in oak, and the efficiency of EAA utilization, which decreased from 80 % in May to <50 % in August. We conclude that the relative nutritional value of red oak and sugar maple for L. dispar is more strongly affected by protein quantity than quality. Surveys of many wild herbaceous species also suggest that leaf-feeding insects would be unlikely to specialize on plants based on protein quality.     

Different mutation patterns of atovaquone resistance to Plasmodium falciparum in vitro and in vivo : rapid detection of codon 268 polymorphisms in the cytochrome b as potential in vivo resistance marker

Background

Anopheles gambiae is the main vector of Plasmodium falciparum in Africa. The mosquito midgut constitutes a barrier that the parasite must cross if it is to develop and be transmitted. Despite the central role of the mosquito midgut in the host/parasite interaction, little is known about its protein composition. Characterisation of An. gambiae midgut proteins may identify the proteins that render An. gambiae receptive to the malaria parasite.

Methods

We carried out two-dimensional gel electrophoresis of An. gambiae midgut proteins and compared protein profiles for midguts from males, sugar-fed females and females fed on human blood.

Results

Very few differences were detected between male and female mosquitoes for the approximately 375 silver-stained proteins. Male midguts contained ten proteins not detected in sugar-fed or blood-fed females, which are therefore probably involved in male-specific functions; conversely, female midguts contained twenty-three proteins absent from male midguts. Eight of these proteins were specific to sugar-fed females, and another ten, to blood-fed females.

Conclusion

Mass spectrometry analysis of the proteins found only in blood-fed female midguts, together with data from the recent sequencing of the An. gambiae genome, should make it possible to determine the role of these proteins in blood digestion or parasite receptivity.     

Antioxidant capacity of flavanols and gallate esters: pulse radiolysis studies

Reactivities of several proanthocyanidins (monomers of condensed tannins) and gallate esters (representing hydrolyzable tannins) with hydroxyl radicals, azide radicals, and superoxide anions were investigated using pulse radiolysis combined with kinetic spectroscopy. We determined the scavenging rate constants and the decay kinetics of the aroxyl radicals both at the wavelength of the semiquinone absorption (275 nm) and the absorption band of the gallate ester ketyl radical (400-420 nm). For most compounds second-order decay kinetics were observed, which reflect disproportionation of the semiquinones. In the case of the oligomeric hydrolysable tannins, pentagalloyl glucose and tannic acid, the decay kinetics were more complex involving sequential first-order and second-order reactions, which could only be resolved by kinetic modeling. A correlation of the reaction rates with hydroxyl radicals (k*OH) with the number of adjacent aromatic hydroxyl groups (i.e., representing catechol and/or pyrogallol structures) was obtained for both condensed and hydrolyzable tannins. Similar correlation for the reactions with azide radicals and superoxide anions are less obvious, but exist as well. We consider proanthocyanidins superior radical scavenging agents as compared with the monomeric flavonols and flavones and propose that these substances rather than the flavonoids proper represent the antioxidative principle in red wine and green tea.     

Tannins from mimosoid legumes of Texas and Mexico

The percentage of tannins in leaves, bark, wood, and immature fruits of several species of Acacia and related mimosoid legumes from the southwestern U.S. and Mexico, along with a few from Costa Rica and Argentina, was determined by a modified hide powder procedure and by precipitation with casein. The relative percentages of hydrolyzable and condensed tannins were determined by the iodate and the vanillin-HCl methods, respectively. Gallotannins of selected samples were also determined by the rhodanine method. Although the amount of total tannins was similar for the first two methods, values for condensed tannins by the vanillin-HCl method were frequently two to four times greater than the total tannin values.     

Effect of defoliation on concentrations of allelochemicals and soluble sugars in leaves of weeping birch (<Emphasis Type="Italic">Betula pendula</Emphasis> roth)

Effect of strong (75%) and complete (100%) artificial defoliation of weeping birch Betula pendula Roth on the dynamics of soluble sugars and phenols—flavonols, catechins, and tannins in leaves of damaged plants was investigated. Within the first 15 days after strong defoliation of birch, no changes were found in leaf contents of flavonol, catechin, and tannin. The concentration of sugars first increased but, on the 10th day after defoliation, it returned to the normal level. One year after strong defoliation, the lead concentrations of catechins and tannins in damaged trees increased, while the concentrations of flavonols and sugars did not differ from that in leaves of control trees. In two years after strong damage, the increased concentration of tannins was retained, while catechins and sugars remained at the control level. One year after complete (100%) artificial defoliation, the leaf concentrations of flavonols and sugars in damaged plants did not differ from that in control plants, while the leaf concentrations of catechins and tannins exceeded those in control plants. Two years after complete damage, the leaves contained an increased amount of tannins, whereas the amounts of catechins, flavonols, and sugars did not differ from the control levels.     

Effects of artificial grassland establishment on soil nutrients and carbon properties in a black-soil-type degraded grassland

Despite a growing knowledge of nutrient limitation for mangrove species and how mangroves adapt to low nutrients, there is scant information about the relative importance of N:P ratio and leaf phenolics variability in determining nutrient conservation. In this study, we evaluated possible nutrient conservation strategies of a mangrove Rhizophora stylosa under nutrient limitation. 1. The leaf nutrient concentrations of R. stylosa changed with season, with the highest N concentration in winter and the highest P concentration in spring for both mature and senescent leaves. Leaf N and P concentrations decreased significantly during leaf senescence. Based on N:P ratios R. stylosa forest was N-limited. Accordingly, the nitrogen resorption efficiency (NRE) was significantly higher than phosphorus resorption efficiency (PRE) for the R. stylosa leaves during leaf senescence. The NRE and PRE both reached the highest in the autumn. Average N and P concentrations in the senescent leaves were 0.15% and 0.06% for R. stylosa, respectively, indicating a complete resorption of N and an incomplete resorption of P. There was a significant negative correlation between nitrogen resorption proficiency (NRP) and NRE, meanwhile phosphorus resorption proficiency (PRP) and PRE correlation was also highly significantly. 2. R. stylosa leaves contained relatively high tannin level. Total phenolics, extractable condensed tannins and total condensed tannins contents increased during leaf senescence, and changed between seasons. The lowest concentrations of total phenolics, extractable condensed tannins and total condensed tannins occurred in summer, total phenolics concentrations were inversely related to nitrogen or phosphorus concentrations. 3. Our results confirmed that resorption efficiency during leaf senescence depends on the type of nutrient limitation, and NRE was much higher than PRE under N-limited conditions. R. stylosa forest developed several nutrient conservation strategies in the intertidal coastline surroundings, including high nitrogen resorption efficiency, low nutrient losses and high tannins level.     

Genetic determination of tannins and herbivore resistance in <Emphasis Type="Italic">Quercus ilex</Emphasis>

Genetic variability of trees influences the chemical composition of tissues. This determines herbivore impact and, consequently, herbivore performance. We evaluated the independent effects of plant genotype and provenance on the tannin content of holm oak (Quercus ilex) and their consequences for herbivory and performance of gypsy moth (Lymantria dispar) larvae. Oak seedlings of 48 open-pollinated families from six populations were grown in a common garden in central Spain. Half the plants were subjected to defoliation by gypsy moth larvae and the other half were destructively sampled for chemical analysis. Tannin content of leaves did not differ significantly among populations but differed significantly among families. Estimates of heritability (h ²) and quantitative genetic differentiation among populations for tannin content (Q _ST) were 0.83 and 0.12, respectively. Defoliation was not related to the tannin content of plants nor to spine and trichome densities of leaves, although positive family–mean associations were observed between defoliation and both seed weight and plant height (P < 0.003). Among the oak populations, differential increase in larval weight gain with defoliation was observed. Leaf tannin content in Q. ilex is genetically controlled but does not influence defoliation or predict performance of the larvae. Different efficiencies of food utilisation depending on the oak genotypes indicate that other plant traits are influencing the feeding patterns and fitness of L. dispar and consequent population dynamics.