Induction of defenses and within-alga variation of palatability in two brown algae from the northern-central coast of Chile: Effects of mesograzers and UV radiation

Macroalgae possess different defense mechanisms in response to herbivory. Some species produce anti-herbivore secondary metabolites, but production of these substances can be costly. Therefore, algae may produce defensive metabolites only in response to herbivory (inducible defense) or defend particular parts of the alga differentially (within-alga variation). In the present study, we examined whether two species of brown algae from the SE-Pacific show evidence of inducible chemical defense (non-polar compounds) or within-alga variation of defense, which we estimated in form of palatability of differently treated algae to amphipod grazers (with live algae and agar-based food containing non-polar algal extracts). In Glossophora kunthii (C. Agardh) J. Agardh, we observed an increase in palatability after algae were acclimated for 12 days without grazers. Subsequent addition of grazers for 12 days then resulted in a reduction of palatability indicating the existence of inducible defense. After removal of grazers for 12 days, these induced effects again disappeared. The reaction of G. kunthii was triggered even by the mere presence of grazers, which suggests that this alga can respond to waterborne cues by reducing palatability. Effects were only found for agar-based food containing non-polar extracts, but not for live algae, suggesting that some parts of the algae are undefended. Our second experiment on within-alga variation confirmed that only apical (growth region) and basal parts (near the holdfast region) of G. kunthii are defended against herbivores. For the second species, Macrocystis integrifolia Bory, the first experiment revealed no induction of defense, while the second experiment on within-alga variation showed that amphipods avoided basal parts and in particular stipes of M. integrifolia but only in live algae. Although both studied algal species differed substantially in their defensive strategies, their reaction was independent of the presence or absence of UV radiation. Thus, it appears that UV effects play only a minor role in anti-herbivore defense, which is in accordance with most previous studies.     

Tetranychus evansi spider mite populations suppress tomato defenses to varying degrees

Plant defense suppression is an offensive strategy of herbivores, in which they manipulate plant physiological processes to increase their performance. Paradoxically, defense suppression does not always benefit the defense‐suppressing herbivores, because lowered plant defenses can also enhance the performance of competing herbivores and can expose herbivores to increased predation. Suppression of plant defense may therefore entail considerable ecological costs depending on the presence of competitors and natural enemies in a community. Hence, we hypothesize that the optimal magnitude of suppression differs among locations. To investigate this, we studied defense suppression across populations of Tetranychus evansi spider mites, a herbivore from South America that is an invasive pest of solanaceous plants including cultivated tomato, Solanum lycopersicum, in other parts of the world. We measured the level of expression of defense marker genes in tomato plants after infestation with mites from eleven different T. evansi populations. These populations were chosen across a range of native (South American) and non‐native (other continents) environments and from different host plant species. We found significant variation at three out of four defense marker genes, demonstrating that T. evansi populations suppress jasmonic acid‐ and salicylic acid‐dependent plant signaling pathways to varying degrees. While we found no indication that this variation in defense suppression was explained by differences in host plant species, invasive populations tended to suppress plant defense to a smaller extent than native populations. This may reflect either the genetic lineage of T. evansi—as all invasive populations we studied belong to one linage and both native populations to another—or the absence of specialized natural enemies in invasive T. evansi populations.     

Pollinator identity and spatial isolation influence multiple paternity in an annual plant

The occurrence and extent of multiple paternity is an important component of variation in plant mating dynamics. However, links between pollinator activity and multiple paternity are generally lacking, especially for plant species that attract functionally diverse floral visitors. In this study, we separated the influence of two functionally distinct floral visitors (hawkmoths and solitary bees) and characterized their impacts on multiple paternity in a self‐incompatible, annual forb, Oenothera harringtonii (Onagraceae). We also situated pollinator‐mediated effects in a spatial context by linking variation in multiple paternity to variation in plant spatial isolation. We documented pronounced differences in the number of paternal sires as function of pollinator identity: on average, the primary pollinator (hawkmoths) facilitated mating with nearly twice as many pollen donors relative to the secondary pollinator (solitary bees). This effect was consistent for both isolated and nonisolated individuals, but spatial isolation imposed pronounced reductions on multiple paternity regardless of pollinator identity. Considering that pollinator abundance and pollen dispersal distance did not vary significantly with pollinator identity, we attribute variation in realized mating dynamics primarily to differences in pollinator morphology and behaviour as opposed to pollinator abundance or mating incompatibility arising from underlying spatial genetic structure. Our findings demonstrate that functionally distinct pollinators can have strongly divergent effects on polyandry in plants and further suggest that both pollinator identity and spatial heterogeneity have important roles in plant mating dynamics.     

Changes in cytokinins are sufficient to alter developmental patterns of defense metabolites in Nicotiana attenuata

Plant defense metabolites are well known to be regulated developmentally. The optimal defense (OD) theory posits that a tssue's fitness values and probability of attack should determine defense metabolite allocations. Young leaves are expected to provide a larger fitness value to the plant, and therefore their defense allocations should be higher when compared with older leaves. The mechanisms that coordinate development with defense remain unknown and frequently confound tests of the OD theory predictions. Here we demonstrate that cytokinins (CKs) modulate ontogeny‐dependent defenses in Nicotiana attenuata. We found that leaf CK levels highly correlate with inducible defense expressions with high levels in young and low levels in older leaves. We genetically manipulated the developmental patterns of two different CK classes by using senescence‐ and chemically inducible expression of CK biosynthesis genes. Genetically modifying the levels of different CKs in leaves was sufficient to alter ontogenic patterns of defense metabolites. We conclude that the developmental regulation of growth hormones that include CKs plays central roles in connecting development with defense and therefore in establishing optimal patterns of defense allocation in plants.     

Variation of chemical compounds in wild Heliconiini reveals ecological factors involved in the evolution of chemical defenses in mimetic butterflies

Evolutionary convergence of color pattern in mimetic species is tightly linked with the evolution of chemical defenses. Yet, the evolutionary forces involved in natural variations of chemical defenses in aposematic species are still understudied. Herein, we focus on the evolution of chemical defenses in the butterfly tribe Heliconiini. These neotropical butterflies contain large concentrations of cyanogenic glucosides, cyanide‐releasing compounds acting as predator deterrent. These compounds are either de novo synthesized or sequestered from their Passiflora host plant, so that their concentrations may depend on host plant specialization and host plant availability. We sampled 375 wild Heliconiini butterflies across Central and South America, covering 43% species of this clade, and quantify individual variations in the different CGs using liquid chromatography coupled with tandem mass spectrometry. We detected new compounds and important variations in chemical defenses both within and among species. Based on the most recent and well‐studied phylogeny of Heliconiini, we show that ecological factors such as mimetic interactions and host plant specialization have a significant association with chemical profiles, but these effects are largely explained by phylogenetic relationships. Our results therefore suggest that shared ancestries largely contribute to chemical defense variation, pointing out at the interaction between historical and ecological factors in the evolution of Müllerian mimicry.     

Host‐associated genomic differentiation in congeneric butterflies: now you see it,now you do not

Ecotypic variation among populations may become associated with widespread genomic differentiation, but theory predicts that this should happen only under particular conditions of gene flow, selection and population size. In closely related species, we might expect the strength of host‐associated genomic differentiation (HAD) to be correlated with the degree of phenotypic differentiation in host‐adaptive traits. Using microsatellite and Amplified Fragment Length Polymorphism (AFLP) markers, and controlling for isolation by distance between populations, we sought HAD in two congeneric species of butterflies with different degrees of host plant specialization. Prior work on Euphydryas editha had shown strong interpopulation differentiation in host‐adapted traits, resulting in incipient reproductive isolation among host‐associated ecotypes. We show here that Euphydryas aurinia had much weaker host‐associated phenotypic differentiation. Contrary to our expectations, we detected HAD in Euphydryas aurinia, but not in E. editha. Even within an E. aurinia population that fed on both hosts, we found weak but significant sympatric HAD that persisted in samples taken 9 years apart. The finding of significantly stronger HAD in the system with less phenotypic differentiation may seem paradoxical. Our findings can be explained by multiple factors, ranging from differences in dispersal or effective population size, to spatial variation in genomic or phenotypic traits and to structure induced by past histories of host‐adapted populations. Other infrequently measured factors, such as differences in recombination rates, may also play a role. Our result adds to recent work as a further caution against assumptions of simple relationships between genomic and adaptive phenotypic differentiation.     

A critical role for Arabidopsis MILDEW RESISTANCE LOCUS O2 in systemic acquired resistance

Members of the MILDEW RESISTANCE LOCUS O (MLO) gene family confer susceptibility to powdery mildews in different plant species, and their existence therefore seems to be disadvantageous for the plant. We recognized that expression of the Arabidopsis MLO2 gene is induced after inoculation with the bacterial pathogen Pseudomonas syringae, promoted by salicylic acid (SA) signaling, and systemically enhanced in the foliage of plants exhibiting systemic acquired resistance (SAR). Importantly, distinct mlo2 mutant lines were unable to systemically increase resistance to bacterial infection after inoculation with P. syringae, indicating that the function of MLO2 is necessary for biologically induced SAR in Arabidopsis. Our data also suggest that the close homolog MLO6 has a supportive but less critical role in SAR. In contrast to SAR, basal resistance to bacterial infection was not affected in mlo2. Remarkably, SAR‐defective mlo2 mutants were still competent in systemically increasing the levels of the SAR‐activating metabolites pipecolic acid (Pip) and SA after inoculation, and to enhance SAR‐related gene expression in distal plant parts. Furthermore, although MLO2 was not required for SA‐ or Pip‐inducible defense gene expression, it was essential for the proper induction of disease resistance by both SAR signals. We conclude that MLO2 acts as a critical downstream component in the execution of SAR to bacterial infection, being required for the translation of elevated defense responses into disease resistance. Moreover, our data suggest a function for MLO2 in the activation of plant defense priming during challenge by P. syringae.     

Host plant and the predator Podisus nigrispinus: when the defense compounds of the plant affect the third trophic level

Many herbivorous insects can overcome chemical plant defenses, using the plant's defensive products for their own good, as a defense against predators. Eucalyptus spp. (Myrtaceae), recently introduced in Brazil, are rich in secondary compounds; however, there are reports that these plants have been suffering from population outbreaks of defoliating Lepidoptera in Brazil. The predator Podisus nigrispinus (Dallas) (Hemiptera: Pentatomidae) has been used against herbivorous insects in eucalyptus plantations, but little is known about its establishment in the field. This study aims to investigate whether the effectiveness of this predator may be affected indirectly by compounds of eucalyptus plants, when compared to guava, Psidium guajava L., a Brazilian native species of Myrtaceae. Thus, we evaluated the performance of P. nigrispinus on larvae of Thyrinteina arnobia (Stoll) (Lepidoptera: Geometridae) reared on eucalyptus (exotic species) or guava plants (native species). Podisus nigrispinus performance (reproduction and survival) was better on larvae fed on guava than on larvae fed on eucalyptus. It is possible that the negative effect on the predator's development occurred because of the plants’ secondary compounds appropriated by caterpillars, due to the short coevolutionary history between eucalyptus and the predator. The data suggest that the chemical compounds that could help the plant's defenses against herbivores may also affect their natural enemies, especially when the interaction between plant and natural enemy involves an exotic plant recently introduced into the insect's habitat.     

Polygyny does not explain the superior competitive ability of dominant ant associates in the African ant‐plant,Acacia (Vachellia) drepanolobium

The Acacia drepanolobium (also known as Vachellia drepanolobium) ant‐plant symbiosis is considered a classic case of species coexistence, in which four species of tree‐defending ants compete for nesting space in a single host tree species. Coexistence in this system has been explained by trade‐offs in the ability of the ant associates to compete with each other for occupied trees versus the ability to colonize unoccupied trees. We seek to understand the proximal reasons for how and why the ant species vary in competitive or colonizing abilities, which are largely unknown. In this study, we use RADseq‐derived SNPs to identify relatedness of workers in colonies to test the hypothesis that competitively dominant ants reach large colony sizes due to polygyny, that is, the presence of multiple egg‐laying queens in a single colony. We find that variation in polygyny is not associated with competitive ability; in fact, the most dominant species, unexpectedly, showed little evidence of polygyny. We also use these markers to investigate variation in mating behavior among the ant species and find that different species vary in the number of males fathering the offspring of each colony. Finally, we show that the nature of polygyny varies between the two commonly polygynous species, Crematogaster mimosae and Tetraponera penzigi: in C. mimosae, queens in the same colony are often related, while this is not the case for T. penzigi. These results shed light on factors influencing the evolution of species coexistence in an ant‐plant mutualism, as well as demonstrating the effectiveness of RADseq‐derived SNPs for parentage analysis.     

Clonal variation in foliar chemistry of aspen: effects on gypsy moths and forest tent caterpillars

Quaking aspen (Populus tremuloides) exhibits striking intraspecific variation in concentrations of phenolic glycosides, compounds that play important roles in mediating interactions with herbivorous insects. This research was conducted to assess the contribution of genetic variation to overall phenotypic variation in aspen chemistry and interactions with gypsy moths (Lymantria dispar) and forest tent caterpillars (Malacosoma disstria). Thirteen aspen clones were propagated from field-collected root material. Insect performance assays, measuring survival, development, growth, and food utilization indices, were conducted with second and/or fourth instars. Leaf samples were assayed for water, nitrogen, total nonstructural carbohydrates, condensed tannins, and phenolic glycosides. Results showed substantial among-clone variation in the performance of both insect species. Chemical analyses revealed significant among-clone variation in all foliar constituents and that variation in allelochemical contents differed more than variation in primary metabolites. Regression analyses indicated that phenolic glycosides were the dominant factor responsible for among-clone variation in insect performance. We also found significant genetic trade-offs between growth and defense among aspen clones. Our results suggest that genetic factors are likely responsible for much of the tremendous phenotypic variation in secondary chemistry exhibited by aspen, and that the genetic structure of aspen populations may play important roles in the evolution of interactions with phytophagous insects. Received: 14 May 1996 / Accepted: 29 January 1997     

Leaf and Soil Nutrients in a Chronosequence of Second‐Growth Forest in Central Amazonia: Implications for Restoration of Abandoned Lands

Subsistence agriculture, cattle ranching, and periodical land abandonment are common land‐use practices in Amazonia. Because changes in land use affect biogeochemical cycles, secondary forests growing after land abandonment develop at varying speeds and spatial patterns, due in part to varying nutrient dynamics. Leaf and soil nutrient concentrations can provide useful information on nutrient cycling processes and strategies of nutrient use by trees that are suitable for introduction to abandoned areas. To understand nutrient dynamics in secondary forests from different regeneration stages, as well as the importance of pioneer species in the regeneration process, we measured the concentration of macronutrients in leaves of three pioneer tree species (Vismia cayennensis, Cecropia sciadophylla, and Bellucia dichotoma) in central Amazon secondary forests. We also measured macronutrients in the topsoil under the trees. We found that type of prior land use, time since abandonment, and number of fire events were significantly correlated with the concentrations of leaf and soil macronutrients, explaining between 10 and 38% of the variation in macronutrient concentrations. The observed patterns suggest that management practices affect the processes involved in N cycling and availability. Of the three pioneer species, C. sciadophylla showed the highest nutrient resorption efficiency and the highest leaf nutritional quality. We suggest that these two features determine a higher potential of C. sciadophylla for natural regeneration and restoration of abandoned lands.     

舍曲林对蚤状溞诱导型反捕食防御代内与代际可塑性的干扰效应

表型可塑性是指生物(尤其是单一基因型)在适应异质环境时表达出不同表型的能力,并且有遗传基础。环境变化调控表型可塑性既可以发生在个体发育进程中(称为代内表型可塑性),也能够以可遗传表型响应的形式持续多个生物世代(称为代际表型可塑性)。浮游动物枝角类常常受到来自鱼类的捕食风险影响而表现出诱导型防御的表型可塑性。诱导型防御的表达在很大程度上受到代谢稳态控制,因此枝角类的诱导型防御的表达容易受到内稳态代谢干扰物(例如抗抑郁药舍曲林)的影响。考虑到舍曲林在水中生物活性高且难以被降解;同时,枝角类世代周期较短,因此需要评估连续多代舍曲林暴露对枝角类反捕食防御代内以及代际可塑性的影响。结果发现：在代内可塑性方面,连续两代的舍曲林暴露放大了鱼类信息素诱导的蚤状溞体长减小趋势以及相对尾刺长度增长趋势,但是对鱼类信息素作用下的种群适合度参数无明显干扰。在代际可塑性方面,随着连续两代舍曲林的浓度上升,亲代经历鱼类信息素产下的子代继续响应鱼类信息素时,体长减小、相对尾刺长度增大,说明形态防御得到加强;同时种群净繁殖量以及种群内禀增长率下降的趋势被放大,由此可能抑制鱼类捕食风险下蚤状溞的种群丰度。上述结果表明,...     

The ITS region of groundwater amphipods: length,secondary structure and phylogenetic information content in Crangonyctoids and Niphargids

The phylogenetic analysis of groundwater amphipods is challenging due to the lack of suitable morphological characters. However, molecular phylogenies based on the 18S and 28S nuclear genes of two Crangonyctoidea species endemic to Iceland, Crymostygius thingvallensis and Crangonyx islandicus, support the taxonomy of these species on the basis of morphological characters. Molecular analyses suggest that the genus Crangonyx is paraphyletic, with the species that is found in Eurasia being highly divergent genetically from the species present in North America and Iceland. Studies of the phylogenetic relationships within the genus Niphargus also warrant further work. The nuclear ITS2 region has recently been proposed as a barcoding marker for plants and animals. In addition, ITS2 has been used to build phylogenies at high taxonomic levels by including its secondary structure. In this study, we want to evaluate the applicability of the ITS region for this group of species and describe its characteristics. The taxonomy of C. thingvallensis, as well as the paraphyly of the genus Crangonyx, is supported herein by phylogenies based on the ITS2 variation. The secondary structure and the length of the ITS2 sequences of the Crangonyctoidea and the Niphargidae species studied are highly variable and are characterized by duplications. The ITS2 sequence of Niphargus plateaui is the longest metazoan sequence deposited in the ITS2 database so far. Although saturation was observed in the nucleotide variation of this marker, the addition of the secondary structure information for the reconstruction of the phylogeny did not add support to the phylogenetic trees. The ITS1 region, which is known to be more variable than ITS2 and bears a large duplication within C. islandicus, was found to be less useful for phylogenetic reconstruction.     

Natural variation in the contribution of microbial density to inducible immune dynamics

Immune responses evolve to balance the benefits of microbial killing against the costs of autoimmunity and energetic resource use. Models that explore the evolution of optimal immune responses generally include a term for constitutive immunity, or the level of immunological investment prior to microbial exposure, and for inducible immunity, or investment in immune function after microbial challenge. However, studies rarely consider the functional form of inducible immune responses with respect to microbial density, despite the theoretical dependence of immune system evolution on microbe‐ versus immune‐mediated damage to the host. In this study, we analyse antimicrobial peptide (AMP) gene expression from seven wild‐caught flour beetle populations (Tribolium spp.) during acute infection with the virulent bacteria Bacillus thuringiensis (Bt) and Photorhabdus luminescens (P.lum) to demonstrate that inducible immune responses mediated by the humoral IMD pathway exhibit natural variation in both microbe density‐dependent and independent temporal dynamics. Beetle populations that exhibited greater AMP expression sensitivity to Bt density were also more likely to die from infection, while populations that exhibited higher microbe density‐independent AMP expression were more likely to survive P. luminescens infection. Reduction in pathway signalling efficiency through RNAi‐mediated knockdown of the imd gene reduced the magnitude of both microbe‐independent and dependent responses and reduced host resistance to Bt growth, but had no net effect on host survival. This study provides a framework for understanding natural variation in the flexibility of investment in inducible immune responses and should inform theory on the contribution of nonequilibrium host‐microbe dynamics to immune system evolution.     

Large‐scale adaptive divergence in Boechera fecunda,an endangered wild relative of Arabidopsis

Many biological species are threatened with extinction because of a number of factors such as climate change and habitat loss, and their preservation depends on an accurate understanding of the extent of their genetic variability within and among populations. In this study, we assessed the genetic divergence of five quantitative traits in 10 populations of an endangered cruciferous species, Boechera fecunda, found in only several populations in each of two geographic regions (WEST and EAST) in southwestern Montana. We analyzed variation in quantitative traits, neutral molecular markers, and environmental factors and provided evidence that despite the restricted geographical distribution of this species, it exhibits a high level of genetic variation and regional adaptation. Conservation efforts therefore should be directed to the preservation of populations in each of these two regions without attempting transplantation between regions. Heritabilities and genetic coefficients of variation estimated from nested ANOVAs were generally high for leaf and rosette traits, although lower (and not significantly different from 0) for water‐use efficiency. Measures of quantitative genetic differentiation, Q_ST, were calculated for each trait from each pair of populations. For three of the five traits, these values were significantly higher between regions compared with those within regions (after adjustment for neutral genetic variation, F_ST). This suggested that natural selection has played an important role in producing regional divergence in this species. Our analysis also revealed that the B. fecunda populations appear to be locally adapted due, at least in part, to differences in environmental conditions in the EAST and WEST regions.     

Genetic factors affecting food‐plant specialization of an oligophagous seed predator

Several ecological and genetic factors affect the diet specialization of insect herbivores. The evolution of specialization may be constrained by lack of genetic variation in herbivore performance on different food‐plant species. By traditional view, trade‐offs, that is, negative genetic correlations between the performance of the herbivores on different food‐plant species favour the evolution of specialization. To investigate whether there is genetic variation or trade‐offs in herbivore performance between different food plants that may influence specialization of the oligophagous seed‐eating herbivore, Lygaeus equestris (Heteroptera), we conducted a feeding trial in laboratory using four food‐plant species. Although L. equestris is specialized on Vincetoxicum hirundinaria (Apocynaceae) to some degree, it occasionally feeds on alternative food‐plant species. We did not find significant negative genetic correlations between mortality, developmental time and adult biomass of L. equestris on the different food‐plant species. We found genetic variation in mortality and developmental time of L. equestris on some of the food plants, but not in adult biomass. Our results suggest that trade‐offs do not affect adaptation and specialization of L. equestris to current and novel food‐plant species, but the lack of genetic variation may restrict food‐plant utilization. As food‐plant specialization of herbivores may have wide‐ranging effects, for instance, on coevolving plant–herbivore interactions and speciation, it is essential to thoroughly understand the factors behind the specialization process. Our findings provide valuable information about the role of genetic factors in food‐plant specialization of this oligophagous herbivore.     

Explosive radiation and spatial expansion across the cold environments of the Old World in an avian family

Our objective was to elucidate the biogeography and speciation patterns in an entire avian family, which shows a complex pattern of overlapping and nonoverlapping geographical distributions, and much variation in plumage, but less in size and structure. We estimated the phylogeny and divergence times for all of the world's species of Prunella based on multiple genetic loci, and analyzed morphometric divergence and biogeographical history. The common ancestor of Prunella was present in the Sino‐Himalayan Mountains or these mountains and Central Asia–Mongolia more than 9 million years ago (mya), but a burst of speciations took place during the mid‐Pliocene to early Pleistocene. The relationships among the six primary lineages resulting from that differentiation are unresolved, probably because of the rapid radiation. A general increase in sympatry with increasing time since divergence is evident. With one exception, species in clades younger than c. 3.7 my are allopatric. Species that are widely sympatric, including the most recently diverged (2.4 mya) sympatric sisters, are generally more divergent in size/structure than allo‐/parapatric close relatives. The distributional pattern and inferred ages suggest divergence in allopatry and substantial waiting time until secondary contact, likely due to competitive exclusion. All sympatrically breeding species are ecologically segregated, as suggested by differences in size/structure and habitat. Colonizations of new areas were facilitated during glacial periods, followed by fragmentation during interglacials—contrary to the usual view that glacial periods resulted mainly in fragmentations.     

Phenological matching rather than genetic variation in host preference underlies geographical variation in host plants used by orange tip butterflies

An insect species that shows variation in host species association across its geographical range may do so either because of local adaptation in host plant preference of the insect or through environmentally or genetically induced differences in the plants, causing variation in host plant suitability between regions. In the present study, we experimentally investigate the host plant preference of Anthocharis cardamines (orange tip butterfly) in two populations from the UK and two from Sweden. Previous reports indicate that A. cardamines larvae are found on different host plant species in different regions of the UK, and some variation has been reported in Sweden. Host plant choice trials showed that females prefer to oviposit on plants in an earlier phenological stage, as well as on larger plants. When controlling for plant phenological stage and size, the host species had no statistically significant effect on the choice of the females. Moreover, there were no differences in host plant species preference among the four butterfly populations. Based on our experiment, the oviposition choice by A. cardamines mainly depends on the phenological stage and the size of the host plant. This finding supports the idea that the geographical patterns of host–plant association of A. cardamines in the UK and Sweden are consequences of the phenology and availability of the local hosts, rather than regional genetic differences in the host species preference of the butterfly.