Phenolic Glycosides in <Emphasis Type="Italic">Populus tremuloides</Emphasis> and their Effects on Long-Term Ungulate Browsing

In the aspen-grassland ecotone of Riding Mountain, Manitoba, lightly browsed vigorous clones of trembling aspen (Populus tremuloides Michx.) occur in close proximity to heavily browsed dieback clones. This study examines whether intraspecific variation in the production of phenolic glycosides is correlated with this strong dichotomy in clonal vigor. Individual clones were sampled over four years at three sites located along a gradient of increasing soil moisture stress. At each site, eight aspen clones of similar size and age were sampled: four vigorous and four dieback clones (total of 24 individual clones). The severity of wapiti (elk) browsing was assessed as the ratio of browse-damaged to total branches per aspen ramet. Statistically significant differences in foliar concentrations of the phenolic glycosides salicortin and tremulacin were observed between vigorous and dieback clones: a mean of 14.8% dry mass for lightly browsed (vigorous) clones, versus just 7.0% for heavily browsed (dieback) clones. Mean concentrations of foliar phenolics were also significantly greater in more moisture-stressed sites. These results demonstrate that the strong dichotomy in clonal vigor (vigorous versus dieback clones) is associated with large differences in phenolic glycoside production. Vigorous clone ramets produce high amounts of phenolic glycosides and have low levels of herbivore browsing and low mortality rates, whereas dieback clone ramets have low amounts of phenolic glycosides and much higher herbivore browsing and mortality rates. This suggests that intraspecific variation in phenolic glycosides in trembling aspen is an important predisposing factor leading to ramet mortality, and by extension to the decline of aspen clones.     

Preserving leaves for tannin and phenolic glycoside analyses: A comparison of methods using three willow taxa

Members of the Salicaceae often produce phenolic glycosides and condensed tannins. There is much debate on the best method for the preservation of leaf material prior to chemical analysis. Published results indicate freeze-drying, a method commonly used for tannin analysis, may be inappropriate for phenolic glycosides, unless done in a manner to prevent thawing during the drying process. Another commonly employed method, air-drying, is appropriate for phenolic glycosides but inappropriate for condensed tannins. I present evidence using willow leaves that demonstrates that: (1) leaves freeze-dried in external flasks without temperature control contain lower concentrations of phenolic glycosides (salicortin and 2-cinnamoylsalicortin), (2) air-dried leaves have reduced concentrations of condensed tannins, while (3) vacuum-dried fresh leaves have high concentrations of both phenolic glycosides and condensed tannins. Freeze-drying caused salicortin and 2-cinnamoyl salicortin concentrations to drop by 20 mg/g and 4 mg/g, respectively. Salicin, a product of salicortin and 2-cinnamoyl salicortin degradation, is absent in vacuum-dried leaves, present in air-dried leaves and very high in freeze-dried leaves. Thus, the presence of salicin in this system is an artifact of the preservation technique. Condensed tannin concentrations dropped nearly 20 mg/g when leaves were air-dried. Thus, vacuum-drying fresh leaves allows researchers to quantify phenolic glycosides and condensed tannins from the same leaf material.     

Induction of Phenolic Glycosides by Quaking Aspen (<Emphasis Type="Italic">Populus tremuloides)</Emphasis> Leaves in Relation to Extrafloral Nectaries and Epidermal Leaf Mining

We studied the effect of epidermal leaf mining on the leaf chemistry of quaking aspen, Populus tremuloides, during an outbreak of the aspen leaf miner, Phyllocnistis populiella, in the boreal forest of interior Alaska. Phyllocnistis populiella feeds on the epidermal cells of P. tremuloides leaves. Eleven days after the onset of leaf mining, concentrations of the phenolic glycosides tremulacin and salicortin were significantly higher in aspen leaves that had received natural levels of leaf mining than in leaves sprayed with insecticide to reduce mining damage. In a second experiment, we examined the time course of induction in more detail. The levels of foliar phenolic glycosides in naturally mined ramets increased relative to the levels in insecticide-treated ramets on the ninth day following the onset of leaf mining. Induction occurred while some leaf miner larvae were still feeding and when leaves had sustained mining over 5% of the leaf surface. Leaves with extrafloral nectaries (EFNs) had significantly higher constitutive and induced levels of phenolic glycosides than leaves lacking EFNs, but there was no difference in the ability of leaves with and without EFNs to induce phenolic glycosides in response to mining. Previous work showed that the extent of leaf mining damage was negatively related to the total foliar phenolic glycoside concentration, suggesting that phenolic glycosides deter or reduce mining damage. The results presented here demonstrate that induction of phenolic glycosides can be triggered by relatively small amounts of mining damage confined to the epidermal tissue, and that these changes in leaf chemistry occur while a subset of leaf miners are still feeding within the leaf.     

Transgenic upregulation of the condensed tannin pathway in poplar leads to a dramatic shift in leaf palatability for two tree-feeding Lepidoptera

Transgenic hybrid aspen (Populus tremula x tremuloides) overexpressing the MYB134 tannin regulatory gene show dramatically enhanced condensed tannin (proanthocyanidin) levels, as well as shifts in other phenolic metabolites. A series of insect bioassays with forest tent caterpillars (Malacosoma disstria) and gypsy moth (Lymantria dispar) caterpillars was carried out to determine how this metabolic shift affects food preference and performance of generalist tree-feeding lepidopterans. Both species showed a distinct preference for the high-tannin MYB134 overexpressor plants, and L. dispar performance was enhanced relative to controls. L. dispar reached greater pupal weight and showed reduced time to pupation when reared on the MYB134 overexpressing poplar. These results were unexpected since enhanced condensed tannin levels were predicted to act as feeding deterrents. However, the data may be explained by the observed decrease in the salicinoids (phenolic glycosides) salicortin and tremulacin that accompanied the upregulation of the condensed tannins in the transgenics. We conclude that for these two lepidopteran species, condensed tannin levels are unlikely to be a major determinant of caterpillar food preference or performance. However, our experiments show that overexpression of a single regulatory gene in transgenic aspen can have a significant impact on herbivorous insects.     

Preservation of salicaceae leaves for phytochemical analyses: Further assessment

The chemistry of the plant family Salicaceae has been of interest to researchers as diverse as chemical ecologists, chemosystematists, and paper chemists. Continuing the debate on proper methods for preservation of plant material prior to analysis, vacuum-drying was recently advocated, because freeze-drying may cause degradation of phenolic glycosides. This study was conducted to clarify the consequences of freeze-drying for foliar secondary chemicals and to evaluate the consequences of vacuum-drying for primary compounds (protein and carbohydrates). Leaves of quaking aspen (Populus tremuloides) were flash-frozen in liquid nitrogen and freeze-dried or vacuum-dried at room temperature. We then analyzed samples for levels of salicortin and tremulacin (phenolic glycosides), condensed tannins, nitrogen, soluble protein, sugars, and starch. Freeze-drying did not alter the concentrations of phenolic glycosides or tannins, relative to vacuum-drying. Freeze-drying did cause a small and inexplicable decline in nitrogen and soluble protein. Vacuum-drying, however, reduced starch concentrations by 38%. We suggest that the vacuum-drying method be used in studies in which carbohydrates are of no interest. For studies measuring carbohydrates, however, freeze-drying is a better alternative, and should effect no changes in levels of secondary compounds if samples are not allowed to thaw during the drying process.     

Chemical model for short-term induction in quaking aspen (Populus tremuloides) foliage against herbivores

Simulated large aspen tortrix (Choristoneura conflictana) herbivory of quaking aspen (Populus tremuloides) induces significant increases in concentrations of two phenol glycosides, salicortin and tremulacin, in leaves within 24 hr. Crushing of leaf tissue, as must occur when aspen leaves are eaten by chewing insects such as the large aspen tortrix, results in conversion of salicortin and tremulacin to 6-hydroxy-2-cyclohexenone (6-HCH). Salicortin, tremulacin, 6-HCH, and its degradation product, catechol, are all toxic to the large aspen tortrix when fed on an artificial diet. These damage-induced chemical changes provide a plausible mechanism for short-term resistance induced in aspen leaves by insect herbivory.     

Clonal Saplings of Trembling Aspen Do Not Coordinate Defense Induction

Induction of plant chemical defenses in response to insect feeding may be localized to the site of damage or expressed systemically, mediated by signal transduction throughout the plant. Such systemic induction processes have been widely investigated in plants with single stems, but rarely in clonal plants comprised of multiple ramets with vascular connections. For a clonal tree species such as trembling aspen (Populus tremuloides Michx), integration of induced defense within clones could be adaptive, as clones are spatially extensive and susceptible to outbreak herbivores. We used pairs of aspen saplings with shared roots, replicated from three genotypes, to determine whether defense-induction signals are communicated within clones. One ramet in each pair was subjected to a damage treatment (feeding by Lymantria dispar, followed by mechanical damage), and subsequent changes in leaf defensive chemistry were measured in both ramets. Responses to damage varied by defense type: condensed tannins (CTs) increased in damaged ramets but not in connected undamaged ramets, whereas salicinoid phenolic glycosides (SPGs) were not induced in any ramets. Genotypes varied in their levels of CTs, but not in their levels of SPGs, and responded similarly to damage treatment. These results suggest that, even with both vascular and volatile information available, young aspen ramets do not induce defenses based on signals or metabolites from other ramets. Thus, unlike other clonal plant species, aspen do not appear to coordinate defense induction within clones. Lack of coordinated early induction in aspen may be related to the function of CTs in tolerance, rather than resistance.     

Secondary chemistry of hybrid and parental willows: Phenolic glycosides and condensed tannins inSalix sericea,S. eriocephala,and their hybrids

Salix sericea andS. eriocephala differ markedly in secondary chemistry.S. sericea produces phenolic glycosides, salicortin and 2-cinnamoylsalicortin, and low concentrations of condensed tannin. In contrast,S. eriocephala produces no phenolic glycosides, but high concentrations of condensed tannins. Hybrid chemistry is intermediate for both types of chemicals, suggesting predominantly additive inheritance of these two defensive chemical systems from the parental species. However, there is extensive variation among hybrids. This variation may be due to genetic variation among parental genotypes, which genes were passed on, or to subsequent back-crossing. The differences in chemistry are likely to exert a strong effect on the relative susceptibility of hybrid and parental willows to herbivores.     

Phytochemical Variation in Quaking Aspen: Effects on Gypsy Moth Susceptibility to Nuclear Polyhedrosis Virus

The performance of gypsy moths (Lymantria dispar) feeding on quaking aspen (Populus tremuloides) is strongly influenced by host foliar chemistry and susceptibility to a nuclear polyhedrosis virus (LdNPV), but the relationship of susceptibility to chemistry is poorly understood. We investigated the effects of genetic and resource-mediated variation in phytochemistry on viral pathogenicity. Trees were grown in pots in a common garden. Disks were punched from aspen leaves, inoculated with LdNPV and fed to third instars. Additional leaves were analyzed for levels of nitrogen, starch, phenolic glycosides, and condensed tannins. Despite marked variation among trees in levels of phenolic glycosides and tannins, we observed minimal variation in larval susceptibility to LdNPV. Viral pathogenicity was only weakly (inversely) correlated with tannin concentrations in one of two experiments. These results suggest that differential defoliation of aspen by gypsy moths in the field is due to the direct effects of host chemistry on larval performance rather than to the indirect effects of host chemistry on efficacy of this natural enemy.     

Browse Quality in Quaking Aspen (Populus tremuloides): Effects of Genotype, Nutrients, Defoliation, and Coppicing

The consequences of interactions among genetic, ontogenetic, and environmental factors for the quality of winter-dormant tissues as food for browsing herbivores is poorly understood. We conducted two sequential common garden studies to assess the impacts of intraspecific genetic variation, nutrient availability, prior defoliation, and ontogenetic stage on the chemical quality of winter-dormant tissue in quaking aspen (Populus tremuloides Michx.). In the first study, saplings of 12 aspen genotypes were grown under low and high soil nutrient conditions, with or without two successive seasons of defoliation. Quantity and quality of current year’s twig growth were assessed. Twig production varied among genotypes and declined under low nutrient availability, but showed little response to prior defoliation. Chemical quality of sapling twigs varied substantially among genotypes, and in response to nutrient availability and prior defoliation. Overall, browse quality improved (nitrogen levels increased while phenolic glycoside and condensed tannin levels decreased) after defoliation. Growth and chemical variables exhibited low to moderate clonal repeatability (broad sense heritability) values. Our second study employed the same 12 genotypes, grown under high-nutrient conditions and with or without two seasons of defoliation. The trees were coppiced to produce root sprouts, which were chemically assessed 1 yr later. Rejuvenation via coppicing led to increased levels of nitrogen, phenolic glycosides (salicortin), and tannins in root sprouts, and the magnitude of change varied among aspen genotypes. Signatures of defoliation nearly 2 yr earlier persisted in terms of elevated levels of phenolic glycosides in root sprouts of previously defoliated trees. Aspen forests likely present browsing herbivores with chemically heterogeneous environments because of the interactions of genetic, ontogenetic, and environmental factors that vary over space and time.     

Interactions between Bacteria And Aspen Defense Chemicals at the Phyllosphere – Herbivore Interface

Plant- and insect-associated microorganisms encounter a diversity of allelochemicals, and require mechanisms for contending with these often deleterious and broadly-acting compounds. Trembling aspen, Populus tremuloides, contains two principal groups of defenses, phenolic glycosides (salicinoids) and condensed tannins, which differentially affect the folivorous gypsy moth, Lymantria dispar, and its gut symbionts. The bacteria genus Acinetobacter is frequently associated with both aspen foliage and gypsy moth consuming that tissue, and one isolate, Acinetobacter sp. R7-1, previously has been shown to metabolize phenolic glycosides. In this study, we aimed to characterize further interactions between this Acinetobacter isolate and aspen secondary metabolites. We assessed bacterial carbon utilization and growth in response to different concentrations of phenolic glycosides and condensed tannins. We also tested if enzyme inhibitors reduce bacterial growth and catabolism of phenolic glycosides. Acinetobacter sp. R7-1 utilized condensed tannins but not phenolic glycosides or glucose as carbon sources. Growth in nutrient-rich medium was increased by condensed tannins, but reduced by phenolic glycosides. Addition of the P450 enzyme inhibitor piperonyl butoxide increased the effects of phenolic glycosides on Acinetobacter sp. R7-1. In contrast, the esterase inhibitor S,S,S,-tributyl-phosphorotrithioate did not affect phenolic glycoside inhibition of bacterial growth. Degradation of phenolic glycosides by Acinetobacter sp. R7-1 appears to alleviate the cytotoxicity of these compounds, rather than provide an energy source. Our results further suggest this bacterium utilizes additional, complementary mechanisms to degrade antimicrobial phytochemicals. Collectively, these results provide insight into mechanisms by which microorganisms contend with their environment within the context of plant-herbivore interactions.     

Effects of genotype, nutrient availability, and defoliation on aspen phytochemistry and insect performance

Genetic and environmental variability, and their interactions, influence phytochemical composition and, in turn, herbivore performance. We evaluated the independent and interactive effects of plant genotype, nutrient availability, and defoliation on the foliar chemistry of quaking aspen (Populus tremuloides) and consequences for performance of gypsy moths (Lymantria dispar). Saplings of four genotypes were grown under two conditions of nutrient availability and subjected to three levels of artificial defoliation. Concentrations of all secondary and primary metabolites evaluated responded to at least one or more of the experimental treatments. Of the secondary metabolites, phenolic glycosides were affected strongly by genotype, less so by nutrient availability, and not induced by defoliation. Condensed tannins were strongly dependent upon genotype, soil nutrient availability, and their interaction, and, in contrast to phenolic glycosides, were induced by artificial defoliation. Of the primary metabolites, foliar nitrogen was affected by genotype and soil nutrient availability. Starch concentrations were affected by genotype, nutrient availability, defoliation and interactions among these factors. Foliar water content responded to genotype, nutrient availability, and defoliation, and the effect of nutrient availability depended on genotype. Herbivore performance on these plants was strongly influenced by plant genotype and soil nutrient availability, but much less so by defoliation. Although several of the compound types (condensed tannins, starch, and water) responded to defoliation, quantitative variation in these compounds did not contribute to substantive changes in herbivore performance. Rather, the primary source of variation in insect performance was due to plant genotype (phenolic glycoside levels), while nutrient availability (foliar nitrogen levels) was of secondary importance. These results suggest that genetic variation in aspen plays a major role in determining patterns of insect performance, whereas environmental variation, such as was tested, here is of negligible importance.     

Root Chemistry in Populus tremuloides: Effects of Soil Nutrients,Defoliation, and Genotype

Although genetic, environmental, and G x E effects on aboveground phytochemistry have been well documented in trembling aspen (Populus tremuloides), little work has focused on the same factors affecting tissues underground. Belowground plant defenses are likely important mediators of root-feeding herbivores that can strongly influence plant fitness. We used a common garden of potted aspen trees to explore the individual and interactive effects of soil nutrient availability, foliar damage, genotype, and their interactions, on concentrations of phytochemicals in aspen roots. Our common garden experiment employed 12 aspen genotypes that were planted into either low- or high-nutrient soil environments. Half of the trees were subjected to defoliation for two successive years, while the others were protected from damage. At the end of the growing season after the second defoliation, we harvested the trees to obtain root samples for which we assessed levels of phenolic glycosides, condensed tannins, nitrogen, and starch. Phenolic glycosides were most affected by genotype, while the other root phytochemicals were most responsive to soil nutrient conditions. The effects of defoliation were observed in interaction with soil nutrient environment and/or genotype. Interestingly, the effect of defoliation on phenolic glycosides was mediated by soil nutrients, whereas the effect of defoliation on condensed tannins was observed in concert with effects of both soil nutrients and genotype. Comparison of data from this study with an earlier, related study revealed that concentrations of phenolic glycosides and condensed tannins are lower in roots than leaves, and less responsive to defoliation. That soil nutrient environment affects root phytochemical concentrations is not unexpected given the intimate association of roots and soil, but the complex interactions between soil nutrients, aboveground damage, and genotype, and their effects on root phytochemistry, are intriguing. Variation in root chemistry could have wide-reaching effects on soil microbial communities, nutrient cycling, and herbivores. Additionally, the response of phytochemicals to damage across organs can link different, spatially separated herbivores as they use different parts of the same plant resource.     

Genotypic Differences and Prior Defoliation Affect Re-Growth and Phytochemistry after Coppicing in <Emphasis Type="Italic">Populus tremuloides</Emphasis>

Although considerable research has explored how tree growth and defense can be influenced by genotype, the biotic environment, and their interaction, little is known about how genotypic differences, prior defoliation, and their interactive effects persist in trees that re-grow after damage that severs their primary stem. To address these issues, we established a common garden consisting of twelve genotypes of potted aspen (Populus tremuloides) trees, and subjected half of the trees to defoliation in two successive years. At the beginning of the third year, all trees were severed at the soil surface (coppiced) and allowed to regenerate for five months. Afterwards, we counted the number of root and stump sprouts produced and measured the basal diameter (d) and height (h) of the tallest ramet in each pot. We collected leaves one and two years after the second defoliation and assessed levels of phenolic glycosides, condensed tannins, and nitrogen. In terms of re-growth, we found that the total number of sprouts produced varied by 3.6-fold among genotypes, and that prior defoliation decreased total sprout production by 24%. The size (d²h) of ramets, however, did not differ significantly among genotypes or defoliation classes. In terms of phytochemistry, we observed genotypic differences in concentrations of all phytochemicals assessed both one and two years after the second defoliation. Two years after defoliation, we observed effects of prior defoliation in a genotype-by-defoliation interaction for condensed tannins. Results from this study demonstrate that genotypic differences and impacts of prior defoliation persist to influence growth and defense traits in trees even after complete removal of above-ground stems, and thus likely influence productivity and plant-herbivore interactions in forests affected by natural disturbances or actively managed through coppicing.     

Resistance and Tolerance of Terminalia sericea Trees to Simulated Herbivore Damage Under Different Soil Nutrient and Moisture Conditions

Resource availability, degree of herbivore damage, genetic variability, and their interactions influence the allocation of investment by plants to resistance and tolerance traits. We evaluated the independent and interactive effects of soil nutrients and moisture, and simulated the effects of herbivore damage on condensed tannins (resistance) and growth/regrowth (tolerance) traits of Terminalia sericea, a deciduous tree in the Kalahari desert that constitutes a major component of livestock diet. We used a completely crossed randomized-block design experiment to examine the effects of nutrients, water availability, and herbivore damage on regrowth and resistance traits of T. sericea seedlings. Plant height, number of branches, internode length, leaf area, leaf mass for each seedling, combined weight of stems and twigs, and root mass were recorded. Condensed tannin concentrations were 22.5 and 21.5% higher under low nutrients and low soil moisture than under high nutrient and high water treatment levels. Tannin concentrations did not differ significantly between control and experimental seedlings 2 mo after simulated herbivore damage. Tannin concentrations correlated more strongly with growth traits under low- than under high-nutrient conditions. No trade-offs were detected among individual growth traits, nor between growth traits and condensed tannins. T. sericea appeared to invest more in both resistance and regrowth traits when grown under low-nutrient conditions. Investment in the resistance trait (condensed tannin) under high-nutrient conditions was minimal and, to a lesser degree, correlated with plant growth. These results suggest that T. sericea displays both resistance and tolerance strategies, and that the degree to which each is expressed is resource-dependent.     

Carbon-nutrient balance hypothesis in within-species phytochemical variation ofSalix lasiolepis

Predictions of the carbon-nutrient balance hypothesis were tested using a study of within-species phytochemical variation in the arroyo willow,Salix lasiolepis. The prediction that a balance between nutrients (total protein) and carbon-based secondary metabolites (total phenols) should exist was supported using water treatment and fertilizer experiments and wild willow clones. Leaf nitrogen content and net photosynthetic rates of plants potted in soil in which parental plants grew was low, indicating that wild plants exist under relatively low nutrient status-high carbon balance conditions. The hypothesis also correctly predicted positive relationships between shoot length and phenols in glasshouse plants, wild plants, and plants in the water treatment experiment and negative relationships between shoot length and phenols in the fertilizer treatment experiment. Total phenolic glycosides, fragilin, picein, salicortin, tremulacin, and tremuloidin all correlated positively with shoot length in glasshouse plants on a carbon-biased balance, and male willows had generally lower levels of phenolic glycosides than females. Salicortin and tremulacin showed the strongest positive relationships with shoot length.     

Effects of large mammalian herbivores and ant symbionts on condensed tannins of Acacia drepanolobium in Kenya

Condensed tannins have been considered to be important inducible defenses against mammalian herbivory. We tested for differences in condensed tannin defenses in Acacia drepanolobium in Kenya over two years among different large mammalian herbivore treatments [total exclusion, antelope only, and megaherbivore (elephants and giraffes) + antelope] and with four different ant symbiont species on the trees. We predicted that (1) condensed tannin concentrations would be lowest in the mammal treatment with the lowest level of herbivory (total exclusion), (2) trees occupied by mutualist ants that protect the trees most aggressively would have lower levels of tannins, and (3) if chemical defense production is costly, there would be a trade-off between tannin concentrations, growth, and mechanical defenses. Mean tannin concentrations increased from total exclusion treatments to wildlife-only treatments to megaherbivore + antelope treatments. In 1997, condensed tannin concentrations were significantly lower in trees occupied by the ant Crematogaster nigriceps, the only ant species that actively removed axillary buds. Contrary to our prediction, trees occupied by ant species that protect the trees more aggressively against mammalian herbivores did not have lower overall levels of condensed tannins. There was no consistent evidence of a trade-off between tannin concentrations and growth rate, but there was a positive correlation between mean thorn length and mean tannin concentrations across species of ant inhabitants and across herbivore treatments in 1997. Contrary to our expectation, trees had higher tannin concentrations in the upper parts of the canopy where there is little herbivory by mammals.     

Rapid Herbivore-Induced Changes in Mountain Birch Phenolics and Nutritive Compounds and Their Effects on Performance of the Major Defoliator, Epirrita autumnata

Insect damage changes plant physiology and chemistry, and such changes may influence the performance of herbivores. We introduced larvae of the autumnal moth (Epirrita autumnataBorkh.) on individual branches of its main host plant, mountain birch (Betula pubescens ssp. czerepanovii (Orlova) Hämet-Ahti) to examine rapid-induced plant responses, which may affect subsequent larval development. We measured systemic responses to herbivory by analyzing chemistry, photosynthesis, and leaf growth, as well as effects on larval growth and feeding, in undamaged branches of damaged and control trees. Larvae reared on leaves from intact branches of the herbivore-damaged trees grew faster than those reared on leaves of control trees, indicating systemic-induced susceptibility. Herbivore damage did not lead to systemic changes in levels of primary nutrients or phenolic compounds. The analyses of photosynthetic activity and individual hydrolyzable tannins revealed a reversal of leaf physiology-herbivore defense patterns. On control trees, consumption by E. autumnata larvae was positively correlated with photosynthetic activity; on damaged trees, this correlation was reversed, with consumption being negatively correlated with photosynthetic activity. A similar pattern was found in the relationship between monogalloylglucose, the most abundant hydrolyzable tannin of mountain birch, and leaf consumption. Among the control trees, consumption was positively correlated with concentrations of monogalloylglucose, whereas among herbivore-damaged trees, this correlation was reversed and became negative. Our results suggest that herbivore performance is related to both concentrations of phenolic compounds and photosynthetic activity in leaves. This linkage between herbivore performance, leaf chemistry, and physiology was sensitive to induced plant responses caused by slight herbivore damage.     

Inheritance patterns of phenolics in F1, F2, and back-cross hybrids of willows: implications for herbivore responses to hybrid plants

The aim of this study was to determine the inheritance pattern of phenolic secondary compounds in pure and hybrid willows and its consequences for plant resistance to leaf-feeding insects. F1, F2, and back-cross hybrids along with pure species were produced by hand pollination of pure, naturally-growing Salix caprea (L., Salicaceae) and S. repens (L.) plants. Leaf concentrations of condensed tannins and seven different phenolic glucosides were determined by using butanol-HCl and HPLC analyses. Insect herbivore leaf damage was measured on the same leaves as used for chemical analyses. We found hybrids to be approximately intermediate between the parental species: S. caprea with high levels of condensed tannins and no phenolic glucosides, and S. repens with low levels of condensed tannins and high levels of phenolic glucosides. We also found a negative correlation between concentrations of condensed tannins and phenolic glucosides, suggesting a trade-off in production of these two substances. F2 hybrids and the hybrid back-crossed to S. caprea were significantly more damaged by insect herbivores than the parental species and the F1 hybrid, indicating reduced resistance and possibly a selective disadvantage for these hybrid categories.