COEVOLUTION BETWEEN HISPANIOLAN CROSSBILLS AND PINE: DOES MORE TIME ALLOW FOR GREATER PHENOTYPIC ESCALATION AT LOWER LATITUDE?

Crossbills (Aves: Loxia) and several conifers have coevolved in predator-prey arms races over the last 10,000 years. However, the extent to which coevolutionary arms races have contributed to the adaptive radiation of crossbills or to any other adaptive radiation is largely unknown. Here we extend our previous studies of geographically structured coevolution by considering a crossbill-conifer interaction that has persisted for a much longer time period and involves a conifer with more variable annual seed production. We examined geographic variation in the cone and seed traits of two sister species of pines, Pinus occidentalis and P. cubensis, on the islands of Hispaniola and Cuba, respectively. We also compared the Hispaniolan crossbill (Loxia megaplaga) to its sister taxa the North American white-winged crossbill (Loxia leucoptera leucoptera). The Hispaniolan crossbill is endemic to Hispaniola whereas Cuba lacks crossbills. In addition and in contrast to previous studies, the variation in selection experienced by these pines due to crossbills is not confounded by the occurrence of selection by tree squirrels (Tamiasciurus and Sciurus). As predicted if P. occidentalis has evolved defenses in response to selection exerted by crossbills, cones of P. occidentalis have scales that are 53% thicker than those of P. cubensis. Cones of P. occidentalis, but not P. cubensis, also have well-developed spines, a known defense against vertebrate seed predators. Consistent with patterns of divergence seen in crossbills coevolving locally with other conifers, the Hispaniolan crossbill has evolved a bill that is 25% deeper than the white-winged crossbill. Together with phylogenetic analyses, our results suggest that predator-prey coevolution between Hispaniolan crossbills and P. occidentalis over approximately 600,000 years has caused substantial morphological evolution in both the crossbill and pine. This also indicates that cone crop fluctuations do not prevent crossbills and conifers from coevolving. Furthermore, because the traits at the phenotypic interface of the interaction apparently remain the same over at least several hundred thousand years, divergence as a result of coevolution is greater at lower latitude where crossbill-conifer interactions have been less interrupted by Pleistocene events.     

The influence of a competitor on the geographic mosaic of coevolution between crossbills and lodgepole pine

The geographic mosaic theory of coevolution posits that the form of selection between interacting species varies across a landscape with coevolution important and active in some locations (i.e., coevolutionary hotspots) but not in others (i.e., coevolutionary coldspots). We tested the hypothesis that the presence of red squirrels (Tamiasciurus hudsonicus) affects the occurrence of coevolution between red crossbills (Loxia curvirostra complex) and Rocky Mountain lodgepole pine (Pinus contorta ssp. latifolia) and thereby provides a mechanism giving rise to a geographic mosaic of selection. Red squirrels are the predominant predispersal seed predator and selective agent on lodgepole pine cones. However, in four isolated mountain ranges east and west of the Rocky Mountains, red squirrels are absent and red crossbills are the main predispersal seed predator. These isolated populations of pine have apparently evolved without Tamiasciurus for about 10,000 to 12,000 years. Based on published morphological, genetic, and paleobotanical studies, we infer that cone traits in these isolated populations that show parallel differences from cones in the Rocky Mountains have changed in parallel. We used data on crossbill and conifer cone morphology and feeding preferences and efficiency to detect whether red crossbills and lodgepole pine exhibit reciprocal adaptations, which would imply coevolution. Cone traits that act to deter Tamiasciurus and result in high ratios of cone mass to seed mass were less developed in the isolated populations. Cone traits that act to deter crossbills include larger and thicker scales and perhaps increased overlap between successive scales and were enhanced in the isolated populations. In the larger, isolated mountain ranges crossbills have evolved deeper, shorter, and therefore more decurved bills to exploit these cones. This provides crossbills with higher feeding rates, and the change in bill shape has improved efficiency by reducing the concomitant increases in body mass and daily energy expenditures that would have resulted if only bill size had increased. These parallel adaptations and counter adaptations in red crossbills and lodgepole pine are interpreted as reciprocal adaptations and imply that these crossbills and pine are in coevolutionary arms races where red squirrels are absent (i.e., coevolutionary hotspots) but not where red squirrels are present (i.e., coevolutionary cold-spots).     

Replicated population divergence caused by localized coevolution? A test of three hypotheses in the red crossbill‐lodgepole pine system

Several lines of evidence support the hypothesis that local populations of red crossbills (Loxia curvirostra complex) enter into a predator-prey arms race with lodgepole pine (Pinus contorta latifolia) in the absence of competing pine squirrels (Tamiasciurus hudsonicus). Nevertheless, the alternative hypotheses that neutral evolution or factors other than squirrels have caused crossbill population differentiation have not been thoroughly tested. We compared crossbill and pine cone morphology between island populations where squirrels are absent or present, and mainland sites where squirrels are present, in order to distinguish among these hypotheses. All comparisons supported an effect of squirrel absence, not island status, on crossbill and cone morphology. Hence our results provide further evidence that strong localized coevolutionary interactions in a geographic mosaic have driven adaptive population differentiation. In addition, vocal differentiation of crossbills was related to the absence of squirrels, but not to island status. As morphological and vocal differentiation is correlated with reproductive isolation in crossbills, the geographic mosaic of coevolution also seems to promote ecological speciation.     

The geographic selection mosaic for ponderosa pine and crossbills: a tale of two squirrels

Recent research demonstrates how the occurrence of a preemptive competitor (Tamiasciurus) gives rise to a geographic mosaic of coevolution for crossbills (Loxia) and conifers. We extend these studies by examining ponderosa pine (Pinus ponderosa), which produces more variable annual seed crops than the conifers in previous studies and often cooccurs with tree squirrels in the genus Sciurus that are less specialized than Tamiasciurus on conifer seed. We found no evidence of seed defenses evolving in response to selection exerted by S. aberti, which was apparently overwhelmed by selection resulting from inner bark feeding that caused many developing cones to be destroyed. In the absence of S. aberti, defenses directed at crossbills increased, favoring larger-billed crossbills and causing stronger reciprocal selection between crossbills and ponderosa pine. However, crossbill nomadism in response to cone crop fluctuations prevents localized reciprocal adaptation by crossbills. In contrast, evolution in response to S. griseus has incidentally defended cones against crossbills, limiting the geographic range of the interaction between crossbills and ponderosa pine. Our results suggest that annual resource variation does not prevent competitors from shaping selection mosaics, although such fluctuations likely prevent fine-scale geographic differentiation in predators that are nomadic in response to resource variability.     

The geographic selection mosaic for squirrels, crossbills and Aleppo pine

The interactions between many species are structured in a geographic mosaic of populations among which selection is divergent. Here we tested the hypothesis that such a geographic selection mosaic arises for common crossbills (Loxia curvirostra) feeding on seeds in the cones of Aleppo pine (Pinus halepensis) because of geographic variation in the occurrence of European red squirrels (Sciurus vulgaris). On the Iberian Peninsula, Sciurus exerted directional selection favouring larger cones with larger scales, which has caused cones there to be larger than in the Balearic Islands where Sciurus are absent. Moreover, cones on the Iberian Peninsula are so large that they are apparently little used by the relatively small-billed crossbills on the Peninsula; selection by Sciurus seems to have made the cones so difficult to feed on that crossbills rely mostly on the seeds of other conifers. Where crossbills are present but Sciurus are absent (Mallorca Island), cones were smaller as a result of relaxation of selection by Sciurus. However, cones on Mallorca had proportionally thicker scales in comparison to where both Sciurus and crossbills are absent (Ibiza Island), presumably as an adaptation against crossbill predation. Here crossbills specialize on Aleppo pine, have relatively large bills and have apparently coevolved in an arms race with Aleppo pine. These results suggest that Sciurus has influenced both the geographic selection mosaics for crossbills and conifers and the adaptive radiation of crossbills in Eurasia much like Tamiasciurus has done in the North America.     

Coevolution between crossbills and black pine: the importance of competitors,forest area and resource stability

Studies of predator‐prey interactions have found that geographically structured coevolution has played an important role in the adaptive diversification of crossbills (Loxia spp.). We extend those studies by considering common crossbills (L. curvirostra) in the Mediterranean where they rely on seeds in the cones of black pine (Pinus nigra). On the continent, where tree squirrels (Sciurus vulgaris) are present, enhanced defenses against crossbills were most evident in larger areas of pine forest. On islands in the absence of tree squirrels, crossbills and black pine have coevolved in a predator‐prey arms race on Cyprus but not Corsica. In contrast to other conifers that island endemic crossbills rely upon, black pine does not hold seeds in its cones year round. Consequently, key to the strong crossbill–pine interaction on Cyprus is likely the presence of an alternative conifer that provides seeds during early summer when black pine seeds are scarce.     

Diversifying Coevolution between Crossbills and Conifers

Coevolution between granivorous crossbills (Loxia spp.) and conifers has been a prominent process in the diversification of crossbills. A striking example occurs in western North America where coevolution between crossbills and Rocky Mountain lodgepole pine (Pinus contorta latifolia) is ongoing in isolated ranges without the crossbill’s dominant competitor for seeds, the red squirrel (Tamiasciurus hudsonicus). Preferential foraging by crossbills on lodgepole pine cones in the South Hills and Albion Mountains, two small mountain ranges in southern Idaho where red squirrels are absent, has led to the evolution of larger, thicker-scaled cones than in nearby ranges where red squirrels are present. This in turn has favored the evolution of larger-billed crossbills that have diverged from other crossbills in the region. However, such diversifying coevolution, resulting from geographic variation in the distribution of strongly interacting species, is vulnerable to species introductions. For example, the introduction of red squirrels caused the precipitous decline and perhaps extinction of the Newfoundland crossbill and perhaps a crossbill endemic to the Cypress Hills, Canada. In general, species introductions act to reduce the geographic variation in species interactions, which may be critical for the diversification of many taxa.     

Interactions among moths, crossbills, squirrels, and lodgepole pine in a geographic selection mosaic

Repeated patterns among biological communities suggest similar evolutionary and ecological forces are acting on the communities. Conversely, the lack of such patterns suggests that similar forces are absent or additional ones are present. Coevolution between a seed predator, the red crossbill (Loxia curvirostra complex), and lodgepole pine (Pinus contorta var. latifolia) exemplifies the ecological and evolutionary predictions for coevolving systems. In the absence of another seed predator and preemptive competitor (pine squirrels Tamiasciurus hudsonicus), natural selection by crossbills results in the evolution of larger cones with thicker distal scales, while relaxation of selection by squirrels results in the evolution of cones with more seeds and a greater ratio of seed mass to cone mass. However, in one range, the Little Rocky Mountains, distal scale thickness has diverged as expected but cone size has not. In these mountains seed predation by lodgepole pine cone borer moths (Eucosma recissoriana) was about 10 times greater than in other ranges lacking squirrels. We quantified moth predation and cone traits and found that moths select for smaller cones with fewer seeds. Thus, selection by moths in the Little Rocky Mountains counters both selection by crossbills for large cone size and relaxation of selection by squirrels favoring more seeds per cone and accounts for the relatively small and few-seeded cones in these mountains. It is also apparent that selection by crossbills changes seed defenses in a manner that favors seed predation by moths, whereas selection by squirrels likely reduces such predation. These results demonstrate the importance of considering the evolutionary consequences of community context in locally evolved (coevolved) traits and interactions.     

A role for habitat area in the geographic mosaic of coevolution between red crossbills and lodgepole pine

Understanding how resource abundance limits adaptive evolution and influences species interactions is an important step towards developing insight into the role of microevolutionary processes in establishing macroevolutionary patterns. We examined how variation in resource abundance (forest area of lodgepole pine Pinus contorta ssp. latifolia) influenced patterns of co-adaptation and coevolution between red crossbill (Loxia curvirostra complex) and lodgepole pine populations. First, we found that crossbill abundance increased logarithmically as forest area increased in mountain ranges lacking a preemptive competitor (pine squirrels Tamiasciurus hudsonicus). Second, seed defences against predation by crossbills increased with increases in crossbill density, suggesting that seed defences have likely evolved in proportion to the intensity of selection that crossbills exert. Third, the average bill size of crossbill populations increased with increasing seed defences, which implies that crossbill offenses increased with increases in seed defences. The large bill size on the largest range is the result of coevolution with lodgepole pine with this crossbill population perhaps speciating. Local adaptation of crossbill populations on smaller ranges, however, is more likely the result of resident crossbills representing a subset of the potential colonists (phenotypic sorting) than of local evolution. In the smallest range, migration and possibly more frequent extinction likely impede local adaptation and may result in maladaptation.     

Temporal variation in breeding and cone size selection by three species of crossbills Loxia spp. in a native Scots pinewood

The breeding biology and cone size selection of crossbills was studied mainly during 1995 to 2002 at Abernethy Forest, Scotland, an ancient native Scots pine Pinus sylvestris wood, where only a single crossbill species, the Scottish crossbill Loxia scotica, was assumed to occur and to be adapted to feed on seeds in Scots pine cones. However, three crossbill species (common Loxia curvirostra, Scottish and parrot crossbills Loxia pytyopsittacus) nested in some years, with the parrot crossbill being the most abundant. Most nests were in old large pines, with the three crossbill species not differing in their use of tree size or stand density for nesting. The mean clutch and brood sizes were 3.8 and 2.9, and their mean survivals were 86 and 74%, respectively, with no significant differences among species. The timing of breeding differed between species, with parrot crossbills breeding earliest (median date 21 March, including second attempts) and common crossbills breeding last (median date 21 April), probably in response to the differing accessibility of Scots pine seeds to these species. The difference in the time of breeding may reduce mixed mating. Crossbills foraged preferentially on trees with small cones when the cones were closed. Small cones had thinner scales than large cones, suggesting that the preference for small cones was related to higher feeding rates on these cones when cones are closed. Such a preference was also found for captive crossbills with the Scottish crossbill showing a more pronounced preference for smaller cones than the larger‐billed parrot crossbill. However, crossbills selected larger cones within trees and trees with larger cones once the cones opened in April. Such a shift occurred presumably because variation in scale thickness has little impact on seed accessibility once cones open, and larger cones have larger and more seeds. The greater ability of parrot crossbills to exploit seeds in closed Scots pine cones allowed parrot crossbills to start breeding earlier and to have young when seeds were most accessible. Only after the cones opened were the smaller‐billed common crossbills able to easily access seeds and to start breeding. The time of breeding of Scottish crossbills was intermediate between common and parrot crossbills, and they probably had an intermediate ability to exploit Scots pine cones. The reason why there were few Scottish crossbills nesting in Abernethy Forest remains a puzzle, considering that native pine wood is assumed to be the ancestral habitat to which the Scottish crossbill is adapted. The breeding season for all crossbills ended in June, when most of the seed from a given cone cohort was shed. This is when starved broods were found, not associated with bad weather.     

Reciprocal selection causes a coevolutionary arms race between crossbills and lodgepole pine

Few studies have shown both reciprocal selection and reciprocal adaptations for a coevolving system in the wild. The goal of our study was to determine whether the patterns of selection on Rocky Mountain lodgepole pine (Pinus contorta spp. latifolia) and red crossbills (Loxia curvirostra complex) were concordant with earlier published evidence of reciprocal adaptations in lodgepole pine and crossbills on isolated mountain ranges in the absence of red squirrels (Tamiasciurus hudsonicus). We found that selection (directional) by crossbills on lodgepole pine where Tamiasciurus are absent was divergent from the selection (directional) exerted by Tamiasciurus on lodgepole pine. This resulted in divergent selection between areas with and without Tamiasciurus that was congruent with the geographic patterns of cone variation. In the South Hills, Idaho, where Tamiasciurus are absent and red crossbills are thought to be coevolving with lodgepole pine, crossbills experienced stabilizing selection on bill size, with cone structure as the agent of selection. These results show that crossbills and lodgepole pine exhibit reciprocal adaptations in response to reciprocal selection, and they provide insight into the traits mediating and responding to selection in a coevolutionary arms race.     

Are the ratios of bill crossing morphs in crossbills the result of frequency-dependent selection?

Summary The direction the lower mandible curves in crossbills (Loxia) is an example of a discrete polymorphism. The lower mandible crosses with equal frequency to the left and to the right in several crossbill populations. I hypothesize that the 1:1 ratio results from negative frequency-dependent selection favouring the rarer morph. A crossbill always orients toward closed conifer cones so that its lower mandible is directed towards the cone axis. Thus, only part of the cone can be reached easily when crossbills have few perch sites and the cone cannot be removed from the branch or otherwise turned round. Since crossbills may visit cones which have previously been foraged on by other individuals, an equal frequency of left-to-right mandible crossings may minimize overlap in the use of cones and enhance foraging efficiency. Experimental data support this hypothesis. Moreover, the variation in the ratio of mandible crossing directions among crossbill populations is consistent with this hypothesis.     

Patterns of genetic variation in the adaptive radiation of New World crossbills (Aves: Loxia)

Incipient species groups or young adaptive radiations such as crossbills (Aves: Loxia) present the opportunity to investigate directly the processes occurring during speciation. New World crossbills include white-winged crossbills (Loxia leucoptera), Hispaniolan crossbills (Loxia megaplaga), and red crossbills (Loxia curvirostra complex), the last of which is comprised of at least nine morphologically and vocally differentiated forms ('call types') where divergent natural selection for specialization on different conifer resources has been strongly implicated as driving diversification. Here we use amplified fragment length polymorphism (AFLP) markers to investigate patterns of genetic variation across populations, call types, and species of New World crossbills. Tree-based analyses using 440 AFLP loci reveal strongly supported clustering of the formally recognized species, but did not separate individuals from the eight call types in the red crossbill complex, consistent with recent divergence and ongoing gene flow. Analyses of genetic differentiation based on inferred allele frequency variation however, reveal subtle but significant levels of genetic differentiation among the different call types of the complex and indicate that between call-type differentiation is greater than that found among different geographic locations within call types. Interpreted in light of evidence of divergent natural selection and strong premating reproductive isolation, the observed genetic differentiation suggests restricted gene flow among sympatric call types consistent with the early stages of ecological speciation.     

When directional selection reduces geographic variation in traits mediating species interactions

Although we often focus on the causes of geographic variation, understanding processes that act to reduce geographic variation is also important. Here, we consider a process whereby adaptive foraging across the landscape and directional selection exerted by a conifer seed predator, the common crossbill (Loxia curvirostra), potentially act to homogenize geographic variation in the defensive traits of its prey. We measured seed predation and phenotypic selection exerted by crossbills on black pine (Pinus nigra) at two sites in the Pindos Mountains, Greece. Seed predation by crossbills was over an order of magnitude higher at the site where cone scale thickness was significantly thinner, which was also the cone trait that was the target of selection at the high predation site. Additional comparisons of selection differentials demonstrate that crossbills exert selection on black pine that is consistent in form across space and time, and increases in strength with increasing seed predation. If predators distribute themselves in relation to the defensive traits of their prey and the strength of selection predators exert is proportional to the amount of predation, then predators may act to homogenize trait variation among populations of their prey in a process analogous to coevolutionary alternation with escalation.     

Divergent selection drives the adaptive radiation of crossbills

Abstract Knowledge of how phenotype influences fitness is necessary if we are to understand the basis of natural selection and how natural selection contributes to adaptive radiations. Here I quantify selection on a wild population of red crossbills ( Loxia curvirostra complex) in the South Hills, Idaho. Bill depth is the target of selection and selection on bill depth is stabilizing. I then show how fitness is related to both bill depth and performance. I use these and previously published relationships to estimate a fitness surface for five species of red crossbills that are part of an ongoing adaptive radiation in western North America. The fitness surface for crossbills has distinct peaks and valleys, with each crossbill species residing on or very near the summits. This work strongly supports a key tenet of the ecological theory of adaptive radiations; namely, divergent selection for utilizing alternative resources is the ultimate cause of adaptive radiations.     

Assortative flocking in crossbills and implications for ecological speciation

How reproductive isolation is related to divergent natural selection is a central question in speciation. Here, we focus on several ecologically specialized taxa or 'call types' of red crossbills (Loxia curvirostra complex), one of the few groups of birds providing much evidence for ecological speciation. Call types differ in bill sizes and feeding capabilities, and also differ in vocalizations, such that contact calls provide information on crossbill phenotype. We found that two call types of red crossbills were more likely to approach playbacks of their own call type than those of heterotypics, and that their propensity to approach heterotypics decreased with increasing divergence in bill size. Although call similarity also decreased with increasing divergence in bill size, comparisons of responses to familiar versus unfamiliar call types indicate that the decrease in the propensity to approach heterotypics with increasing divergence in bill size was a learned response, and not a by-product of calls diverging pleiotropically as bill size diverged. Because crossbills choose mates while in flocks, assortative flocking could lead indirectly to assortative mating as a by-product. These patterns of association therefore provide a mechanism by which increasing divergent selection can lead to increasing reproductive isolation.     

CAN SELECTION BY AN ECTOPARASITE DRIVE A POPULATION OF RED CROSSBILLS FROM ITS ADAPTIVE PEAK?

The bill structures of different call types of red crossbills (Loxia curvirostra complex) in western North America usually approximate the predicted optima for foraging on single species of conifers. One clear exception is the call type in the South Hills, Idaho, that is coevolving in an evolutionary arms race with Rocky Mountain lodgepole pine (Pinus contorta ssp. latifolia). Although South Hills crossbills forage only on the cones of these lodgepole pines, their average bill depth is smaller than that predicted to be optimal. Because preliminary data showed that large-billed males were more likely to exhibit symptoms of ectoparasitic mite (Knemidokoptes jamaicensis) infestation, the goal of our study was to further quantify the incidence of mite infestation and determine whether selection by mites may have favored smaller-billed crossbills and thus driven crossbills away from the foraging optimum. We estimated annual survival of both infected and uninfected South Hills crossbills using program MARK, which allows for auxiliary variables such as bill size and sex to be included in survival analyses. Mite infestation depressed crossbill survival and, especially for males, caused directional selection against larger-billed individuals. Such selection may explain why South Hills crossbills have smaller bills than the optimum and why average bill size for males has decreased from 1998 to 2003. This selection may also explain why the degree of sexual size dimorphism has decreased by nearly 50% since 1998.     

Vocal types in crossbill populations (Loxia spp.) of Southwest Europe

The evolution of crossbills is one of the most fascinating topics in evolutionary ecology. Recent studies have shown an astonishing divergence in terms of vocalisation between morphologically quite similar crossbill populations in the Red/Common Crossbill complex (Loxia curvirostra) of North America and Europe. Some evidence even indicates the existence of “cryptic” species with different vocal types and bill sizes, which are adapted to different conifer species. However, there is so far no strong genetic evidence for the existence of separate species, although assortative mating occurs with respect to bill size. To understand the role of vocalisation in the speciation process of crossbill taxa, basic studies that assess the distribution of vocal types of crossbills and the use of different habitats and resources are needed. In our study, we investigated the occurrence of crossbill vocal types in Southwest Europe. In addition to the well-known vocal types described first by Robb (Dutch Birding 22:61–107, 2000) for the Benelux and Great Britain, we discovered at least six more vocal types in the Mediterranean area. Some vocal types were found exclusively in rather small areas, e.g. in the Pyrenees, the Sierra de Cazorla, Sierra de Javalambre and on Corsica, and appeared to be tightly linked to certain habitat types and pine species. Overall, vocal types in the Mediterranean had a more local occurrence than vocal types from northern populations, which were more widely distributed. This might reflect the nomadic behaviour of northern European crossbills, which feed, in contrast to Mediterranean crossbills, mostly on rather unstable food sources, especially spruce seeds. Furthermore, the vocal types of Mediterranean crossbills show at least some similarities to the vocal types of the rather sedentary crossbills of North Europe (L. pytyopsittacus, L. scotica), which are as well adapted to pine seeds. This might reflect a common ancestry of crossbills adapted for pines. We therefore suggest the existence of two main groups of crossbills in Europe: one group that is rather sedentary and feeds mainly on pine seeds (L. pytyopsittacus, L. scotica and the Mediterranean forms), and another group in Central, Northern and Eastern Europe that is highly nomadic and mostly feeds on spruce seeds (L. curvirostra). Further studies are needed to unravel the consistency of vocal types and the genetic relationship between the different forms, and to provide more evidence for the degree of assortative mating of crossbills with distinct vocalisation breeding in sympatry.     

Beak morphology predicts apparent survival of crossbills: due to selective survival or selective dispersal?

Dozens of morphologically differentiated populations, subspecies and species of crossbills (genus Loxia) exist. It has been suggested that this divergence is due to variation in the conifer cones that each population specialises upon, requiring a specific beak size to efficiently separate the cone scales. If so, apparent survival should depend on beak size. To test this hypothesis, we undertook multievent capture–recapture modelling for 6844 individuals monitored during 27 years in a Pyrenean common crossbill L. curvirostra population in a forest of mountain pine Pinus uncinata. Apparent survival was indeed related to beak width, resulting in stabilizing selection around an optimum that was close to the observed mean beak width, indicating that local crossbill beak morphology is adapted to the conifer they feed upon. Both natural selection (selective mortality) and selective emigration of maladapted individuals may explain our findings. As is often the case in capture–recapture analyses but rarely recognised, we could not formally decompose apparent survival into selective mortality versus selective permanent emigration. Nonetheless, there are several indications that selective permanent emigration should not be fully excluded. First, natural selection by itself would have to be unusually strong compared to other empirical estimates to create the observed pattern of apparent survival. Second, the observed mean beak width was a bit lower than the estimated optimum beak width. This can be explained by immigration of crossbills with smaller beaks originating from southern populations, which may subsequently have left the study area permanently in response to low food intake. This is in line with a detected transient effect in the data, yet apparently little influx from crossbills from northern Europe. When permanent emigration is phenotypically selective this will have ecological and evolutionary consequences, so this possibility deserves more attention in general.     

Assortative mating and patterns of inheritance indicate that the three crossbill taxa in Scotland are species

There are three breeding taxa of crossbills in highland Scotland, the common crossbill Loxia curvirostra , parrot crossbill L. pytyopsittacus and endemic Scottish crossbill L. scotica . These taxa show no genetic differentiation in neutral DNA (microsatellites and mitochondrial DNA), but do show some differentiation in morphology and have distinct calls. Mating patterns and bill size inheritance were examined to test four different hypotheses for their genetic similarity: (1) the three taxa belong to the same species and differences in bill sizes are caused by phenotypic plasticity, (2) the taxa are a result of genetic polymorphism at a single locus, (3) the taxa largely act as species but occasionally cross-breed, such that gene flow results in genetic homogenisation of part of the genome, and (4) the taxa are fully reproductively isolated species but diverged only recently, such that insufficient time has passed for neutral DNA to differentiate by genetic drift. Hypotheses 1 and 2 could be excluded because wild crossbills mated assortatively with respect to bill depth (a measure of bill size) and bill depth was highly heritable; narrow-sense heritabilities of 0.58 for female and 0.71 for male Scottish crossbills. Only two of 46 pairs (4.3%) were not assortative by bill size. Pairing was strongly assortative by flight calls (50 out of 52 pairs, 96.2%) and excitement calls (88 out of 93 pairs, 94.6%). Therefore, the three crossbill taxa do not cross-mate freely, nor do they seem totally reproductively isolated (excluding hypothesis 4). Thus, the results mostly favour hypothesis 3. The small amount of gene flow among crossbill taxa may contribute to the lack of genetic differentiation in neutral DNA. However, this appears to be insufficient not to classify them as species, as long as the calls can be regarded as diagnostic.