Ecological and Evolutionary Stochasticity Shape Natural Selection.

The distribution of biodiversity depends on the combined and interactive effects of ecological and evolutionary processes. The joint contribution of these processes has focused almost exclusively on deterministic effects, even though mechanisms that increase the importance of random ecological processes are expected to also increase the importance of random evolutionary processes. Here we manipulate the sizes of old field fragments to generate correlated sampling effects for a focal population (a gall maker) and its enemy community. Traits and communities were more variable in smaller patches. However, because of the preference of some enemies for some trait values (gall sizes), random variation in population mean trait values exacerbated differences in community composition. The random distribution of traits and interactions created predictable but highly variable patterns of natural selection. Our study highlights how stochastic processes can affect ecological and evolutionary processes structuring the strength and direction of selection locally and at larger scales.

Individual and Population Differences Shape Species Interactions and Natural Selection.

Trait variation is central to our understanding of species interactions, and trait variation arising within species is increasingly recognized as an important component of community ecology. Ecologists generally consider intraspecific variation either among or within populations, yet these differences can interact to create patterns of species interactions. These differences can also affect species interactions by altering processes occurring at distinct scales. Specifically, intraspecific variation may shape species interactions simply by shifting a population's position along a trait-function map or by shifting the relationship between traits and their ecological function. I test these ideas by manipulating within- and among-population intraspecific variation in wild populations of a gall-forming insect before quantifying species interactions and phenotypic selection. Within- and among-population differences in gall size interact to affect attack rates by an enemy community, but among-population differences were far more consequential. Intraspecific differences shaped species interactions by both shifting the position of populations along the trait-function map and altering the relationship between traits and their function, with ultimate consequences for patterns of natural selection. I suggest that intraspecific variation can affect communities and natural selection by acting through individual- and population-level mechanisms.

Consumer-resource interactions along urbanization gradients drive natural selection.

Urbanization is an important component of global change. Urbanization affects species interactions, but the evolutionary implications are rarely studied. We investigate the evolutionary consequences of a common pattern: the loss of high trophic-level species in urban areas. Using a gall-forming fly, Eurosta solidaginis, and its natural enemies that select for opposite gall sizes, we test for patterns of enemy loss, selection, and local adaptation along five urbanization gradients. Eurosta declined in urban areas, as did predation by birds, which preferentially consume gallmakers that induce large galls. These declines were linked to changes in habitat availability, namely reduced forest cover in urban areas. Conversely, a parasitoid that attacks gallmakers that induce small galls was unaffected by urbanization. Changes in patterns of attack by birds and parasitoids resulted in stronger directional selection, but loss of stabilizing selection in urban areas, a pattern which we suggest may be general. Despite divergent selective regimes, gall size did not very systematically with urbanization, suggesting but not conclusively demonstrating that environmental differences, gene flow, or drift, may have prevented the adaptive divergence of phenotypes. We argue that the evolutionary effects of urbanization will have predictable consequences for patterns of species interactions and natural selection.

Adaptive Divergence in a Defense Symbiosis Driven from the Top Down.

Most studies of adaptive radiation in animals focus on resource competition as the primary driver of trait divergence. The roles of other ecological interactions in shaping divergent phenotypes during such radiations have received less attention. We evaluate natural enemies as primary agents of diversifying selection on the phenotypes of an actively diverging lineage of gall midges on tall goldenrod. In this system, the gall of the midge consists of a biotrophic fungal symbiont that develops on host-plant leaves and forms distinctly variable protective carapaces over midge larvae. Through field studies, we show that fungal gall morphology, which is induced by midges (i.e., it is an extended phenotype), is under directional and diversifying selection by parasitoid enemies. Overall, natural enemies disruptively select for either small or large galls, mainly along the axis of gall thickness. These results imply that predators are driving the evolution of phenotypic diversity in symbiotic defense traits in this system and that divergence in defensive morphology may provide ecological opportunities that help to fuel the adaptive radiation of this genus of midges on goldenrods. This enemy-driven phenotypic divergence in a diversifying lineage illustrates the potential importance of consumer-resource and symbiotic species interactions in adaptive radiation.

Keystone Individuals Alter Ecological and Evolutionary Consumer-Resource Dynamics.

Intraspecific variation is central to our understanding of evolution and ecology, but these fields generally consider either the mean trait value or its variance. Alternatively, the keystone individual concept from behavioral ecology posits that a single individual with an extreme phenotype can have disproportionate and irreplaceable effects on group dynamics. Here, I generalize this concept to include nonbehavioral traits and broader ecological and evolutionary dynamics. I test for the effects of individuals with extreme phenotypes on the ecology and evolution of a gall-forming fly and its natural enemies that select for opposite gall sizes. Specifically, I introduce a putatively keystone predator-attracting individual gall-maker, hypothesizing that the presence of such an individual should (1) increase gall maker population-level mortality, (2) cause consumer communities to be dominated by species that are most attracted to the keystone individual, (3) increase selection for traits conferring defense against the most common consumer, and (4) weaken patterns of stabilizing selection. I find support for both the ecological and evolutionary consequences of single individuals with extreme phenotypes, suggesting that they can be considered keystone individuals. I discuss the generality of the keystone individual concept, suggesting likely consequences for ecology and evolution.

Identification of an insect-produced olfactory cue that primes plant defenses.

It is increasingly clear that plants perceive and respond to olfactory cues. Yet, knowledge about the specificity and sensitivity of such perception remains limited. We previously documented priming of anti-herbivore defenses in tall goldenrod plants (Solidago altissima) by volatile emissions from a specialist herbivore, the goldenrod gall fly (Eurosta solidaginis). Here, we explore the specific chemical cues mediating this interaction. We report that E,S-conophthorin, the most abundant component of the emission of male flies, elicits a priming response equivalent to that observed for the overall blend. Furthermore, while the strength of priming is dose dependent, plants respond even to very low concentrations of E,S-conophthorin relative to typical fly emissions. Evaluation of other blend components yields results consistent with the hypothesis that priming in this interaction is mediated by a single compound. These findings provide insights into the perceptual capabilities underlying plant defense priming in response to olfactory cues.Plants are able to prime anti-herbivore defenses in response to olfactory cues of insect pests. Here, Helms et al. identify the insect pheromone E,S-conophthorin produced by the goldenrod gall fly as the specific chemical component that elicits this priming response in goldenrod plants.

[Effects of Cuscuta australis parasitism on the growth, reproduction and defense of Solidago canadensis].

In order to find out how parasitic Cuscuta australis influences the growth and reproduction of Solidago canadensis, the effects of the parasitism of C. australis on the morphological, growth and reproductive traits of S. canadensis were examined and the relationships between the biomass and the contents of the secondary metabolites were analyzed. The results showed that the parasitism significantly reduced the plant height, basal diameter, root length, root diameter, root biomass, stem biomass, leaf biomass, total biomass, number of inflorescences branches, axis length of inflorescence, and number of inflorescence. In particular, plant height, number of inflorescence and the stem biomass of parasitized S. canadensis were only 1/2, 1/5 and 1/8 of non-parasitized plants, respectively. There was no significant difference of plant height, root length, stem biomass and total biomass between plants parasitized with high and low intensities. But the basal diameter, root volume, leaf biomass, root biomass, the number of inflorescences branches, axis length of inflorescence and number of inflorescence of S. canadensis parasitized with high intensity were significantly lower than those of plants parasitized with low intensity. The parasitism of C. australis significantly increased the tannins content in the root and the flavonoids content in the stem of S. canadensis. The biomass of S. canadensis was significantly negatively correlated with the tannin content in the root and the flavonoids content in the stem. These results indicated that the parasitism of C. australis could inhibit the growth of S. canadensis by changing the resources allocation patterns as well as reducing the resources obtained by S. canadensis.

Calcium signaling mediates cold sensing in insect tissues.

The ability to rapidly respond to changes in temperature is a critical adaptation for insects and other ectotherms living in thermally variable environments. In a process called rapid cold hardening (RCH), insects significantly enhance cold tolerance following brief (i.e., minutes to hours) exposure to nonlethal chilling. Although the ecological relevance of RCH is well-established, the underlying physiological mechanisms that trigger RCH are poorly understood. RCH can be elicited in isolated tissues ex vivo, suggesting cold-sensing and downstream hardening pathways are governed by brain-independent signaling mechanisms. We previously provided preliminary evidence that calcium is involved in RCH, and here we firmly establish that calcium signaling mediates cold sensing in insect tissues. In tracheal cells of the freeze-tolerant goldenrod gall fly, Eurosta solidaginis, chilling to 0 °C evoked a 40% increase in intracellular calcium concentration as determined by live-cell confocal imaging. Downstream of calcium entry, RCH conditions significantly increased the activity of calcium/calmodulin-dependent protein kinase II (CaMKII) while reducing phosphorylation of the inhibitory Thr306 residue. Pharmacological inhibitors of calcium entry, calmodulin activation, and CaMKII activity all prevented ex vivo RCH in midgut and salivary gland tissues, indicating that calcium signaling is required for RCH to occur. Similar results were obtained for a freeze-intolerant species, adults of the flesh fly, Sarcophaga bullata, suggesting that calcium-mediated cold sensing is a general feature of insects. Our results imply that insect tissues use calcium signaling to instantly detect decreases in temperature and trigger downstream cold-hardening mechanisms.

Rampant host- and defensive phenotype-associated diversification in a goldenrod gall midge.

Natural selection can play an important role in the genetic divergence of populations and their subsequent speciation. Such adaptive diversification, or ecological speciation, might underlie the enormous diversity of plant-feeding insects that frequently experience strong selection pressures associated with host plant use as well as from natural enemies. This view is supported by increasing documentation of host-associated (genetic) differentiation in populations of plant-feeding insects using alternate hosts. Here, we examine evolutionary diversification in a single nominal taxon, the gall midge Asteromyia carbonifera (O.S.), with respect to host plant use and gall phenotype. Because galls can be viewed as extended defensive phenotypes of the midges, gall morphology is likely to be a reflection of selective pressures by enemies. Using phylogenetic and comparative analyses of mtDNA and nuclear sequence data, we find evidence that A. carbonifera populations are rapidly diversifying along host plant and gall morphological lines. At a broad scale, geography explains surprisingly little genetic variation, and there is little evidence of strict co-cladogenesis with their Solidago hosts. Gall morphology is relatively labile, distinct gall morphs have evolved repeatedly and colonized multiple hosts, and multiple genetically and morphologically distinct morphs frequently coexist on a single host plant species. These results suggest that Asteromyia carbonifera is in the midst of an adaptive radiation driven by multitrophic selective pressures. Similar complex community pressures are likely to play a role in the diversification of other herbivorous insect groups.

Differences in spatial distribution, morphology, and communities of herbivorous insects among three cytotypes of Solidago altissima (Asteraceae).

PREMISE OF THE STUDY: Polyploidy in plants can result in genetic isolation, ecological differences among cytotypes, and, ultimately, speciation. Cytotypes should be sympatric only if they are segregated in an ecological niche or through prezygotic isolation. We tested whether sympatric diploid, tetraploid, and hexaploid ramets of Solidago altissima L. (Asteraceae) differ in their ecological niche. METHODS: We measured how cytotypes were distributed within habitats, their morphology, and the composition of their communities of herbivorous insects at 10 natural field sites. We also conducted a common garden experiment to confirm whether observed differences in morphology or communities of herbivores were due to cytotype or environmental effects. KEY RESULTS: Diploid ramets often grew in open areas, relatively far from woody plants, and were associated with a high species richness of herbaceous plants, especially grasses. Hexaploids often grew in heavy shading under woody plants where grasses were scarce. Finally, tetraploids usually grew in transition areas between diploids and hexaploids. Hexaploid ramets also were taller than ramets of the other cytotypes and had larger leaves. Two species of insects, the leaf-galling fly Asteromyia carbonifera and the phloem-tapping aphid Uroleucon nigrotuberculatum, were more abundant on hexaploid ramets than on ramets of other cytotypes in the field. When grown in a common garden, however, cytotypes were similar in morphology and communities of herbivores. CONCLUSIONS: We conclude that cytotypes of S. altissima differ in their spatial distribution within habitats and that spatial variation in environmental factors influence plant morphology and communities of herbivorous insects.

A gall-inducing caterpillar species increases essential fatty acid content of its host plant without concomitant increases in phytohormone levels.

Gall-inducing insects are accomplished plant parasites that can profoundly influence host-plant physiology. We recently reported that the caterpillar Gnorimoschema gallaesolidaginis failed to significantly alter emissions of host-plant volatiles that often recruit natural enemies of insect herbivores, and demonstrated that a caterpillar species feeding on linolenate-deficient plant tissues avoids inducing some of the indirect defenses of its host plant. Here, we investigate whether absence of volatile responses to the galler G. gallaesolidaginis could similarly be explained by a lack of linolenate in galls. We screened interior and exterior tissue of galls and control stems of Solidago altissima for free linolenate, linoleate, 12-oxo-phytodienoate, jasmonate, and salicylate. We found, unexpectedly, that G. gallaesolidaginis strongly increased amounts of linolenic and linoleic acids inside galls without associated increases in two downstream products, 12-oxo-phytodienoic or jasmonic acid. In contrast, the generalist caterpillar Heliothis virescens induced elevated levels of linolenic, linoleic, 12-oxo-phytodienoic, and jasmonic acids in S. altissima. Moreover, these two fatty acids and 12-oxo-phytodienoate were significantly and positively associated with jasmonic acid, suggesting that increased levels of these precursors can lead directly to greater amounts of jasmonic acid. Taken together, these findings suggest that gall insects may be able to nutritionally enhance their food source without inducing concomitant increases in phytohormones and associated defense responses.

Plant genetics shapes inquiline community structure across spatial scales.

Recent research in community genetics has examined the effects of intraspecific genetic variation on species diversity in local communities. However, communities can be structured by a combination of both local and regional processes and to date, few community genetics studies have examined whether the effects of instraspecific genetic variation are consistent across levels of diversity. In this study, we ask whether host-plant genetic variation structures communities of arthropod inquilines within distinct habitat patches--rosette leaf galls on tall goldenrod (Solidago altissima). We found that genetic variation determined inquiline diversity at both local and regional spatial scales, but that trophic-level responses varied independently of one another. This result suggests that herbivores and predators likely respond to heritable plant traits at different spatial scales. Together, our results show that incorporating spatial scale is essential for predicting the effects of genetically variable traits on different trophic levels and levels of diversity within the communities that depend on host plants.

Behavioural, ecological and genetic evidence confirm the occurrence of host-associated differentiation in goldenrod gall-midges.

Host-associated differentiation (HAD) is considered a step towards ecological speciation and an important mechanism promoting diversification in phytophagous insects. Although the number of documented cases of HAD is increasing, these still represent only a small fraction of species and feeding guilds among phytophagous insects, and most reports are based on a single type of evidence. Here we employ a comprehensive approach to present behavioural, morphological, ecological and genetic evidence for the occurrence of HAD in the gall midge Dasineura folliculi (Diptera: Cecidomyiidae) on two sympatric species of goldenrods (Solidago rugosa and S. gigantea). Controlled experiments revealed assortative mating and strong oviposition fidelity for the natal-host species. Analysis of mitochondrial DNA showed an amount of genetic divergence between the two host-associated populations compatible with cryptic species rather than host races. Lower levels of within-host genetic divergence, gall development and natural-enemy attack in the S. gigantea population suggest this is the derived host.

Host-race formation: promoted by phenology, constrained by heritability.

Host-race formation is promoted by genetic trade-offs in the ability of herbivores to use alternate hosts, including trade-offs due to differential timing of host-plant availability. We examined the role of phenology in limiting host-plant use in the goldenrod gall fly (Eurosta solidaginis) by determining: (1) whether phenology limits alternate host use, leading to a trade-off that could cause divergent selection on Eurosta emergence time and (2) whether Eurosta has the genetic capacity to respond to such selection in the face of existing environmental variation. Experiments demonstrated that oviposition and gall induction on the alternate host, Solidago canadensis, were the highest on young plants, whereas the highest levels of gall induction on the normal host, Solidago gigantea, occurred on intermediate-age plants. These findings indicate a phenological trade-off for host-plant use that sets up the possibility of divergent selection on emergence time. Heritability, estimated by parent-offspring regression, indicated that host-race formation is impeded by the amount of genetic variation, relative to environmental, for emergence time.

Gall insects can avoid and alter indirect plant defenses.

Parasitic species can dramatically alter host traits. Some of these parasite-induced changes can be considered adaptive manipulations that benefit the parasites. Gall-inducing insects are parasites well known for their ability to alter host-plant morphology and physiology, including the distribution of plant defensive compounds. Here it was investigated whether gall-inducing species alter indirect plant defenses, involving the release of volatile compounds that are attractive to foraging natural enemies. Using field and factorial laboratory experiments, volatile production by goldenrod (Solidago altissima) plants was examined in response to attack by two gall-inducing species, the tephritid fly Eurosta solidaginis and the gelechiid moth Gnorimoschema gallaesolidaginis, as well as the meadow spittlebug, Philaenus spumarius, and the generalist caterpillar Heliothis virescens. Heliothis virescens elicited strong indirect defensive responses from S. altissima, but the gall-inducing species and spittlebugs did not. More significantly, infestation by E. solidaginis appeared to suppress volatile responses to subsequent attack by the generalist caterpillar. The extensive control that E. solidaginis apparently exerts over host-plant defense responses may reduce the predation risk for the gall inducer and the subsequent herbivore, and could influence community-level dynamics, including the distribution of herbivorous insect species associated with S. altissima parasitized by E. solidaginis.

Aquaporins play a role in desiccation and freeze tolerance in larvae of the goldenrod gall fly, Eurosta solidaginis.

Survival of freezing not only requires organisms to tolerate ice formation within their body, but also depends on the rapid redistribution of water and cryoprotective compounds between intra- and extracellular compartments. Aquaporins are transmembrane proteins that serve as the major pathway through which water and small uncharged solutes (e.g. glycerol) enter and leave the cell. Consequently, we examined freeze-tolerant larvae of the goldenrod gall fly, Eurosta solidaginis, to determine whether aquaporins are present and if their presence promotes freeze tolerance of specific tissues. Immunoblotting with mammalian anti-AQP2, -AQP3 and -AQP4 revealed corresponding aquaporin homologues in E. solidaginis, whose patterns of expression varied depending on acclimation temperature and desiccation treatment. To examine the role of aquaporins in freeze tolerance, we froze fat body, midgut and salivary gland tissues in the presence and absence of mercuric chloride, an aquaporin inhibitor. Survival of fat body and midgut cells was significantly reduced when mercuric chloride was present. In contrast, survival of the salivary gland did not decrease when it was frozen with mercuric chloride. Overall, this study supports our hypothesis that naturally occurring aquaporins in E. solidaginis are regulated during desiccation and promote cell survival during freezing.

Ducking as a means of resistance to herbivory in tall goldenrod, Solidago altissima.

Many populations of goldenrod show a peculiar, genetically controlled stem dimorphism. In Solidago altissima, for instance, while most stems are erect, a sizable minority (the "candy-cane" stems) nod at the apex during growth. We used data from three studies to test the hypothesis that this candy-cane growth form confers resistance to herbivory. In a controlled growth trial, we showed that nodding is a temporary phenomenon that coincides with the oviposition period of at least two common apex-attacking herbivores: the tephritid galler Eurosta solidaginis and the gall midge Rhopalomyia solidaginis. In a large field survey, stems of candy-cane genets were only half as likely to be ovipunctured by E. solidaginis. In a common-garden study, candy-cane stems were less likely to be ovipunctured by E. solidaginis, and they were galled only half as often by R. solidaginis as erect stems. These results suggest that the candy-cane stems of goldenrod possess a resistance strategy that allows them to essentially duck and hide from certain herbivores.

Good mothers, bad mothers, and the nature of resistance to herbivory in Solidago altissima.

Evidence of poor correspondence between an insect herbivore's oviposition preferences and the performance of its offspring has generally been attributed either to maladaptive behavior of the insect mother or inadequate measurement by the researcher. In contrast, we hypothesize that many cases of "bad mothers" in herbivores may be a byproduct of the hierarchical way natural selection works on resistance in host plants. Epistatic selection on the components of resistance (i.e., antixenosis and antibiosis) may generate negative genetic correlations between the resistance components, which could counteract the efforts of herbivores to oviposit on the best hosts for the performance of their offspring. In common garden and greenhouse experiments, we measured aspects of antixenosis and antibiosis resistance in 26 genets of tall goldenrod, Solidago altissima, against two common herbivores: the gall-inducing fly Eurosta solidaginis and the spittlebug Philaenus spumarius. Goldenrod antixenosis and antibiosis were positively correlated against E. solidaginis and negatively correlated against P. spumarius. Analogously, population-wide preference-performance correlations were positive for the gall flies and negative for the spittlebugs. Several natural history differences between the two insects could make gall flies better mothers, including better synchrony of the phenologies of the flies and the host plant, the much narrower host range of the gall flies than the spittlebugs, and the more sedentary lifestyle of the gall fly larvae than the spittlebug nymphs. If these results are typical in nature, then negative genetic correlations in antixenosis and antibiosis in plants may often result in zero or negative population-wide correlations between preference and performance in herbivores, and thus may be an important reason why herbivorous insects often appear to be bad mothers.

Differential attack on diploid, tetraploid, and hexaploid Solidago altissima L. by five insect gallmakers.

Genetic variation among plants can influence host choice and larval performance in insect herbivores. Ploidy (cytotype) variation is a particularly dramatic form of plant genetic variation, and where diploid and polyploid cytotypes of a species occur in sympatry, they may provide herbivores with choices that are distinguished by profound and genome-wide genetic differences. We tested for non-random attack by five gallmaking insect herbivores on diploid, tetraploid, and hexaploid cytotypes of the goldenrod Solidago altissima L., working in seven midwestern US populations where the ploidies co-occur on spatial scales relevant to insect host choice. For four of the five herbivores, attack was non-random with respect to ploidy at one or more sites. Ploidy effects on attack were complex: the ploidy subjected to highest attack varied both across herbivores within sites and (for most herbivores) across sites within herbivores. Ploidy effects on attack will alter rates of encounter between insect herbivores-either increasing or decreasing the likelihood of two herbivores sharing a host plant ramet, compared with the case with no effects of ploidy. Plant ploidy variation appears likely to have a major impact on insect community organization, and perhaps on plant-herbivore coevolution, but that impact is likely to be spatially heterogeneous.