Biogeography of Alaska paper birch (Betula neoalaskana): latitudinal patterns in chemical defense and plant architecture.

The latitudinal herbivory-defense hypothesis (LHDH) predicts that plants near the equator will be more heavily defended against herbivores than are plants at higher latitudes. Although this idea is widely found in the literature, recent studies have called this biogeographic pattern into question. We sought to evaluate the LHDH in a high-latitude terrestrial ecosystem where fire and mammalian herbivores may contribute to selection for higher levels of defensive chemistry. To address this objective, we collected seeds of Alaska paper birch (Betula neoalaskana) from nine locations along two north-south transects between 55 degrees N and 62 degrees N latitudes in western, interior Canada. The birch seeds were planted in pots in a common garden in Madison, Wisconsin, USA. From the resulting seedlings, we determined levels of chemical defense by assessing the density of resin glands, which have been shown to be negatively correlated with browsing. To assess plant architectural traits such as height, mean individual leaf area, and root-to-shoot ratio, we harvested a subset of the birch seedlings. Further, we used these traits to examine growth-defense trade-offs. Contrary to the LHDH, we found a positive correlation between chemical defense and latitude. Investigating relationships with fire, we found a strong positive correlation between resin gland density and percentage of area annually burned (PAAB) around each collection location and also between PAAB and latitude. Additionally, birch seedlings originating from higher latitudes were shorter, smaller-leaved, and rootier than their lower-latitude counterparts. Growth-defense trade-offs were observed in negative correlations between resin gland density and height and leaf size. Seedlings with higher resin gland densities also allocated less biomass to shoots and more to roots. These results further call into question the LHDH and provide specific information about latitudinal trends in plant defense at high, northern latitudes where fire is a major ecosystem driver and mammals are the main herbivores of deciduous trees such as birches. We propose that these interconnected relationships are the key drivers of the positive correlation between defense and latitude in B. neoalaskana. Understanding patterns of boreal plant defense and growth is especially important because high latitude ecosystems are particularly susceptible to climate change. Key words: Alaska paper birch; Betula neoalaskana; biogeography; chemical defense; grotvth-defense trade-offs; inherent growth rate; latitude; latitudinal herbivory-defense hypothesis; papyr'feric acid; plant ar- chitecture; resin glands; root-to-shoot ratio.

Modeling the dynamics of woody plant-herbivore interactions with age-dependent toxicity.

In this paper we study the effects that woody plant chemical defenses may have on interactions between boreal hares that in winter feed almost entirely on twigs. We focus particularly on the fact that toxin concentration often varies with the age of twig segments. The model incorporates the fact that the woody internodes of the youngest segments of the twigs of the deciduous angiosperm species that these hares prefer to eat are more defended by toxins than the woody internodes of the older segments that subtend and support the younger segments. Thus, the per capita daily intake of the biomass of the older segments of twigs by hares is much higher than their intake of the biomass of the younger segments of twigs. This age-dependent toxicity of twig segments is modeled using age-structured model equations which are reduced to a system of delay differential equations involving multiple delays in the woody plant-hare dynamics. A novel aspect of the modeling was that it had to account for mortality of non-consumed younger twig segment biomass when older twig biomass was bitten off and consumed. Basic mathematical properties of the model are established together with upper and lower bounds on the solutions. Necessary and sufficient conditions are found for the linear stability of the equilibrium in which the hare is extinct, and sufficient conditions are found for the global stability of this equilibrium. Numerical simulations confirmed the analytical results and demonstrated the existence of limit cycles over ranges of parameters reasonable for hares browsing on woody vegetation in boreal ecosystems. This showed that age dependence in plant chemical defenses has the capacity to cause hare-plant population cycles, a new result.

Dynamics of the volatile defense of winter "dormant" balsam poplar (Populus balsamifera).

6-Hydroxycylohex-2-en-1-one (6-HCH) has been reported as a major chemical defense of the winter-dormant internodes of balsam poplar (Populus balsamifera) against feeding by herbivores such as the snowshoe hare (Lepus americanus). We report that the concentration of 6-HCH in the fall internodes is triggered by a single hard frost, and then undergoes an exponential decline through volatilization over the winter that results in barely detectable quantities by early spring. We conclude that the role of 6-HCH in the defense of mature balsam poplar is more complex than simply acting as a toxin. Rather, 6-HCH's role as a defensive agent must evolve over the course of the winter from being a co-toxin to a cue for a conditioned flavor aversion (CFA) to finally having no role by late spring.

Fire drives transcontinental variation in tree birch defense against browsing by snowshoe hares.

Fire has been the dominant disturbance in boreal America since the Pleistocene, resulting in a spatial mosaic in which the most fire occurs in the continental northwest. Spatial variation in snowshoe hare (Lepus americanus) density reflects the fire mosaic. Because fire initiates secondary forest succession, a fire mosaic creates variation in the abundance of early successional plants that snowshoe hares eat in winter, leading to geographic variation in hare density. We hypothesize that fire is the template for a geographic mosaic of natural selection: where fire is greatest and hares are most abundant, hare browsing has most strongly selected juvenile-phase woody plants for defense. We tested the hypothesis at multiple spatial scales using Alaska birch (Betula neoalaskana) and white birch (Betula papyrifera). We also examined five alternative hypotheses for geographic variation in antibrowsing defense. The fire-hare-defense hypothesis was supported at transcontinental, regional, and local scales; alternative hypotheses were rejected. Our results link transcontinental variation in species interactions to an abiotic environmental driver, fire. Intakes of defense toxins by Alaskan hares exceed those by Wisconsin hares, suggesting that the proposed selection mosaic may coincide with a geographic mosaic of coevolution.

Papyriferic acid, an antifeedant triterpene from birch trees, inhibits succinate dehydrogenase from liver mitochondria.

Papyriferic acid (PA) is a triterpene that is secreted by glands on twigs of the juvenile ontogenetic phase of resin producing tree birches (e.g., Betula neoalaskana, B. pendula) and that deters browsing by mammals such as the snowshoe hare (Lepus americanus). We investigated the pharmacology of PA as a first step in understanding its antifeedant effect. After oral administration to rats, PA and several metabolites were found in feces but not urine, indicating that little was absorbed systemically. Metabolism involved various combinations of hydrolysis of its acetyl and malonyl ester groups, and hydroxylation of the terpene moiety. The presence of a malonyl group suggested a possible interaction with succinate dehydrogenase (SDH), a mitochondrial enzyme known to be competitively inhibited by malonic acid. The effect of PA on the oxidation of succinate by SDH was examined in mitochondrial preparations from livers of ox, rabbit, and rat. In all three species, PA was a potent inhibitor of SDH. Kinetic analysis indicated that, unlike malonate, PA acted by an uncompetitive mechanism, meaning that it binds to the enzyme-substrate complex. The hydrolysis product of PA, betulafolienetriol oxide, was inactive on SDH. Overall, the evidence suggests that PA acts as the intact molecule and interacts at a site other than the succinate binding site, possibly binding to the ubiquinone sites on complex II. Papyriferic acid was potent (K(iEIS) ranged from 25 to 45 microM in the three species) and selective, as malate dehydrogenase was unaffected. Although rigorous proof will require further experiments, we have a plausible mechanism for the antifeedant effect of PA: inhibition of SDH in gastrointestinal cells decreases mitochondrial energy production resulting in a noxious stimulus, 5-HT release, and sensations of nausea and discomfort. There is evidence that the co-evolution of birches and hares over a large and geographically-diverse area in Northern Europe and America has produced marked differences in the formation of PA by birches, and the tolerance of hares to dietary PA. The present findings on the metabolic fate and biochemical effects of PA provide a rational basis for investigating the mechanisms underlying differences among populations of hares in their tolerance of a PA-rich diet.

Troublesome toxins: time to re-think plant-herbivore interactions in vertebrate ecology.

Earlier models of plant-herbivore interactions relied on forms of functional response that related rates of ingestion by herbivores to mechanical or physical attributes such as bite size and rate. These models fail to predict a growing number of findings that implicate chemical toxins as important determinants of plant-herbivore dynamics. Specifically, considerable evidence suggests that toxins set upper limits on food intake for many species of herbivorous vertebrates. Herbivores feeding on toxin-containing plants must avoid saturating their detoxification systems, which often occurs before ingestion rates are limited by mechanical handling of food items. In light of the importance of plant toxins, a new approach is needed to link herbivores to their food base. We discuss necessary features of such an approach, note recent advances in herbivore functional response models that incorporate effects of plant toxins, and mention predictions that are consistent with observations in natural systems. Future ecological studies will need to address explicitly the importance of plant toxins in shaping plant and herbivore communities.

Instability of the snowshoe hare and woody plant interaction.

We show that Alaskan woody plants respond to browsing in two ways that might destabilize a plantherbivore interaction and account for snowshoe hare population 'cycles'. (1) Browse production of preferred, earliersuccessional woody plants increases in response to moderate levels of browsing. Such yield increases are potentially destabilizing. Later successional woody plants show decreases in yield after moderate browsing, which is consistent with the persistence of snowshoe hares in late successional 'refuge' habitats (Keith 1966, Wolff 1980). (2) Many woody plants are destructively overbrowsed or girdled at the peak of the snowshoe hare cycle. The more palatable and plastic, early to mid successional plants respond by sprouting accompanied by juvenile reversion. Sprouts are markedly less palatable than mature shoots. We show here that sprout palatability and twig biomass are restored in 2-3 years for earlier successional plants, but palatability may not recover for 4-10 years in sprouts of some mid to late successional plants. The decrease in palatability helps to account for the snowshoe hare 'crash' (assuming that damage to more palatable plants is widespread during the 'peak'), and the 2-3 year time lag for recovery of more palatable species could account for (May 1974) the observed 8-11 year period of the hare cycles. Browse yield increases acting during the snowshoe hare population nadir and increase, and sprouting with juvenile reversion acting during the hare peak and decline can in principle account for the oscillatory nature and the observed 8-11 year periodicity of the snowshoe hare cycle.

Effects of above-ground browsing by mammals on mycorrhizal infection in an early successional taiga ecosystem.

Using an exclosure experiment in the willow stage of primary succession on the floodplain of the Tanana River, we tested the hypothesis that browsing can reduce mycorrhizal infection. We measured the effects winter browsing by moose (Alcesalces) and snowshoe hare (Lepusamericanus) had on mycorrhizal infection and fine root biomass of willow (Salix spp.) and balsam poplar (Populusbalsamifera). We found that protection from winter browsing increased ectomycorrhizal infection by 10% in the top 5 cm of the soil profile, by 23% at 5-10 cm, and by 42% at the 10-15 cm depth. Mammal browsing in taiga forests is now recognized as a major cause of the shift from palatable deciduous species such as willow and balsam poplar to less palatable species such as alder and spruce. We suggest that browsing-induced reduction in ectomycorrhizal infection of salicaceous species plays a central role in this shift in plant community composition.

Foliage acceptability to browsing ruminants in relation to seasonal changes in the leaf chemistry of woody plants in a South African savanna.

We investigated seasonal changes in food selection by hand-reared kudus and impalas in savanna vegetation in northern Transvaal, South Africa. The acceptability of the leaves of woody plants to these animals was compared with leaf concentrations of nutrients, fibre components and old leaf phenophases. No consistently significant correlation was found between acceptability and any single chemical factor. Based on an a priori palatability classification, discriminant function analysis separated relatively palatable species from unpalatable species in terms of a linear combination of protein and condensed tannin concentrations. The high acceptability of certain otherwise unpalatable species during the new leaf phenophase was related to elevation of protein levels relative to condensed tannin contents. Species were added to the diet during the dry season approximately in the order of their relative protein-condensed tannin difference.

Lack of induced chemical defense in juvenile Alaskan woody plants in response to simulated browsing.

Juvenile individuals of five species of Alaskan trees and shrubs were clipped at six levels of intensity and sampled for resin, tannin, nutrient, and carbohydrate content. Clipping caused no induction of putative defensive compounds (resins or tannins), indicating that the high level of resin found in many Alaskan woody plants in response to browsing is best explained as a reversion to a juvenile growth stage rather than an induction of a specific chemical defense.

Defense of winter-dormant Alaska paper birch against snowshoe hares.

Mature growth-phase internodes of Alaska paper birch (Betula resinifera) are preferred by the snowshoe hare (Lepus americanus) over juvenile growth-phase internodes due to the low food value of the latter. While the mature over juvenile preferencec cannot be explained by the levels of inorganic nutrients or gross chemical fractions (resins or phenols), it can be explained by the striking differences in secondary metabolites of the two growth phases. The principle compound which renders the juvenile phase internodes unpalatable is papyriferic acid, a triterpene which is a demonstrated feeding deterrent to snowshoe hares and which is present in juvenile internodes at concentrations 25 times greater than those in mature internodes.