An analysis of population genetic differentiation and genotype–phenotype association across the hybrid zone of carrion and hooded crows using microsatellites and MC1R

The all black carrion crow ( Corvus corone corone ) and the grey and black hooded crow ( Corvus   corone cornix ) meet in a narrow hybrid zone across Europe. To evaluate the degree of genetic differentiation over the hybrid zone, we genotyped crows from the centre and edges of the zone, and from allopatric populations in northern (Scotland–Denmark–Sweden) and southern Europe (western–central northern Italy), at 18 microsatellites and at a plumage candidate gene, the MC1R gene. Allopatric and edge populations were significantly differentiated on microsatellites, and populations were isolated by distance over the hybrid zone in Italy. Single-locus analyses showed that one locus, CmeH9, differentiated populations on different sides of the zone at the same time as showing only weak separation of populations on the same side of the zone. Within the hybrid zone there was no differentiation of phenotypes at CmeH9 or at the set of microsatellites, no excess of heterozygotes among hybrids and low levels of linkage disequilibrium between markers. We did not detect any association between phenotypes and nucleotide variation at MC1R , and the two most common haplotypes occurred in very similar frequencies in carrion and hooded crows. That we found a similar degree of genetic differentiation between allopatric and edge populations irrespectively of their location in relation to the hybrid zone, no differentiation between phenotypes within the hybrid zone, and neither heterozygote excess nor consistent linkage disequilibrium in the hybrid zone, is striking considering that carrion and hooded crows are phenotypically distinct and sometimes recognised as separate species.     

Genetic variation of carrion and hooded crows and their hybrids based on RAPD-PCR data

RAPD-PCR analysis of genetic variability of carrion crows, hooded crows and their phenotypic hybrids from the zones of overlap of their areals and hybridization of parental forms in Siberia has been carried out. According to RAPD variability the following parameters were estimated for both species and their hybrids: genetical distances (DN), polymorphism (P95), mean expected heterozygosity (H(e)), gene fixation coefficient (Fst) and the portion of interpopulation differentiation (Gst). It was shown that the rate of genetic variability in carrion crows and hooded crows is less than in the hybrids. It turned out that the hybrids were genetically closer to each other (DN = 0.295) than to both parental species (DN = 0.441 and 0.397 with C. corone and C. cornix, correspondingly). The studied individuals of C. cornix show the minimal level of heterozygosity and polymorphism (H(e) = 0.12 and P95 = = 27.8%) in comparison with C. corone and the crows of hybrid zone (H(e) = 0.18 and 0.20; P95 = 42.6% and 50.4%, correspondingly). Right's indexes of inbreeding (Fst = 0.285) and population spliting (Gst = 0.352) indicate low genetic variability in all the analysed birds. Nevertheless UPGMA and NJ dendrogrammes differentiate the individuals from the areals of carrion crows, hooded crows and from hybrid zone to different clusters. Our data correlate with molecular-genetic investigations of Corvidae carried out with other methods, and do not contradict the conception of carrion crows and hooded crows as semispecies in the context of the conception of overspecies.     

Habitat preferences and positive assortative mating in an avian hybrid zone

The relatively narrow hybrid zone between hooded and carrion crow is characterised by a high proportion of breeding hybrids, no known intrinsic postzygotic costs, and most likely, high gene flow between populations. It is therefore of interest to identify factors preventing a breakdown of the zone. In this study, we investigate the possibility that habitat choice could create assortative mating in the German–Danish part of the zone. In a GIS analysis, we used two approaches to investigate the role of habitat selection in the maintenance of this zone. We tested if there are 1) any correlations between habitat and the shape of the hybrid zone, and 2) any differences in habitat preferences between territorial individuals of the crow phenotypes. The most extensive habitat, non‐irrigated arable land (Habitat 1), which covers almost 50% of the area in the zone was preferred by all three phenotypes (carrion, hooded and hybrid crows), but showed no change in frequency across the zone. Pastures (Habitat 2) and agricultural areas mixed with natural vegetation (Habitat 4) correlated with the shape of the zone, but only H4 differed in preference between the crow phenotypes. H4 was preferred by hybrids and hooded crows, but not by carrion crows. This habitat is present only in 17% of the hooded crow territories, and thus, it is not likely to have a strong influence on the maintenance of the hybrid zone. There was evidence of assortative mating in pure phenotypes, whereas hybrids showed no consistent mating pattern. Because our GIS analyses suggest that species specific habitat preferences are not involved in shaping the assortativeness, we discuss other possible mechanisms.     

Hybrid zone maintenance by non-adaptive mate choice

In a spatial simulation we show that formation of a stable, narrow hybrid zone requires no selection, only that individuals can recognize their own population type since this is a prerequisite for assortative mating. The European crow occurs in two species/subspecies that meet in a long hybrid zone, the black carrion crow Corvus corone and the grey and black hooded crow C. cornix. In a previous study (Brodin A, Haas F (2006) Anim Behav 72:139) we mimicked sexual imprinting in nestling crows with artificial neural networks, using a learning process that simulates proximate retina perception. The networks were trained on 700 images to recognize either carrion, hooded or hybrid phenotypes as their own. After training the networks were exposed to 300 new, unfamiliar pictures of crows. The networks that had been trained on pure subspecies then showed strong preference for their own type. Networks trained on hybrid crows showed weaker preference for their own type. Assuming that these preferences will determine mate choice preferences we have combined them with empirical data on dispersal of young crows to investigate what kind of geographical distribution pattern this would create. We then assume that the two subspecies meet and hybridize along a straight border. In only 60 generations a stable, narrow hybrid zone that resembles the real hybrid zone was formed. This zone remained stable over time. With minor adjustments of parameters the results would not only fit the width but also the shapes of the clines in the field. An erratum to this article can be found at     

Extra‐pair paternity as a strategy to reduce the costs of heterospecific reproduction? Insights from the crow hybrid zone

Within hybrid zones of socially monogamous species, the number of mating opportunities with a conspecific can be limited. As a consequence, individuals may mate with a heterospecific (social) partner despite possible fitness costs to their hybrid offspring. Extra‐pair copulations with a conspecific may thus arise as a possible post hoc strategy to reduce the costs of hybridization. We here assessed the rate of extra‐pair paternity in the hybrid zone between all‐black carrion crows (Corvus (corone) corone) and grey hooded crows (C. (c.) cornix) and tested whether extra‐pair paternity (EPP) was more likely in broods where parents differed in plumage colour. The proportion of broods with at least one extra‐pair offspring and the proportion of extra‐pair offspring were low overall (6.98% and 2.90%, respectively) with no evidence of hybrid broods having higher EPP rates than purebred nests.     

Aprocta matronensis in crows (Corvus corone corone) from northern Italy.

Aprocta matronensis (Nematoda, Spirurida) has been found in the orbital cavities of carrion crows. Hooded crows and crows from the hybrid crow zone in the studied areas were not infected. The parasite population showed the typical morphology described for this species. This is the first record of A. matronensis in crows in Italy and in Corvus corone corone. Some hypotheses about the distribution of the parasites in crow populations are discussed.     

Gene‐flow across the European crow hybrid zone – a spatial simulation

In hybrid zones populations that are otherwise allopatric meet and produce hybrids. One of the most well‐known hybrid zones occurs throughout much of Europe between the hooded and carrion crows. Even though these species (or subspecies) of crows look very different, genetic differentiation is weak, and introgression seems to be common. In a spatial simulation that was based on empirically estimated values, we investigated how introgressing alleles that would confer some fitness advantage would flow across the zone. The advantage was assumed to be some unknown factor that enhanced survival for carriers relative to non‐carriers. We varied the yearly survival advantage between 0 to 10% and recorded how this would affect zone shape and position. In the simulation we cycled ‘yearly’ events such as reproduction, mortality and juvenile dispersal. We started the simulation by equipping all individuals of one crow type outside the zone with a homozygotic set of the beneficial allele. At all levels of advantage the allele first rapidly became fixed in the donator crow type, then transgressed into the receiving type more slowly and finally became fixed in all individuals. The time until fixation varied from around 176 yr for a 10% advantage to around 20 000 yr for 0% advantage. An exciting discovery was that the position of the zone would move during the introgression but then stop. The reason is that the beneficial allele would be common in the donating type but not in the receiving type during the introgression event. At large the crow hybrid zone appears to have been stable for a long time but there have been small recent northward movements of carrion crows in both Scotland and Denmark. We suggest that introgression by itself is a factor that should be considered as an explanation for why hybrid zones move temporarily.     

The exclusion of avian predators from aggregations of nesting lapwings (Vanellus vanellus)

Territorial lapwings in Aberdeenshire, Scotland, largely prevented carrion crows, (Corvus corone), their main egg predators, from approaching close to nest-sites and from entering the area within an aggregation of 11 nests. Predation rates of artifical nests were significantly lower when placed within 10 m of active lapwing nests than when>200 m from nest-sites, indicating that the presence of lapwings afforded protection from crow predation. Further artificial nest experiments showed that the amount of protection declined linearly with increasing distance from the nest-site, and some degree of protection occurred to at least 30–50 m from it. The size of this protected zone was apparently related to the number of lapwing nests present, and the survival time of eggs in artificial nests reflected the lapwings' nesting density. The defended zones around each lapwing nest overlapped appreciably in dense or large nesting aggregations, leading to the observed protection by generally excluding crows, from the nesting area.     

Mesopredators constrain a top predator: competitive release of ravens after culling crows

Although predator control programmes rarely consider complex competitive interactions among predators, it is becoming clear that removal of larger ‘superior’ competitors often releases the ‘inferior’ ones and can precipitate trophic cascades. In contrast, our study indicates that culling hooded crows Corvus cornix appears to release a larger competitor, the common raven Corvus corax. Ravens ranged more widely, and the predation of artificial nests was significantly faster (although total predation was similar), after the removal of crows. Our study provides evidence of a novel reversal of competitive release where a larger species was freed from constraints imposed on its distribution and behaviour by a smaller species, and emphasizes the importance of considering community and ecosystem effects of predator manipulations when undertaken for conservation or game management.     

THE MAINTENANCE OF A CLINE IN THE MARINE SNAIL LITTORINA SAXATILIS: THE ROLE OF HOME SITE ADVANTAGE AND HYBRID FITNESS

Steep clinal transitions in one or several inherited characters between genetically distinct populations are usually referred to as hybrid zones. Essentially two different mechanisms may maintain steep genetic clines. Either selection acts against hybrids that are unfit over the entire zone due to their mixed genetic origin (endogenous selection), or hybrids and parental types attain different fitness values in different parts of the cline (exogenous selection). Survival rate estimates of hybrids and parental forms in different regions of the cline may be used to distinguish between these models to assess how the cline is maintained. We used reciprocal transplants to test the relative survival rates of two parental ecotypes and their hybrids over microscale hybrid zones in the direct-developing marine snail Littorina saxatilis (Olivi) on the rocky shores of Galicia, Spain. One of the parental forms occupies upper and the other lower shores, and the hybrids are found at various proportions (1–38%) along with both parental forms in a midshore zone a few meters wide. The survival rate over one month was 39-52% of the native ecotype on upper shores, but only 2-8% for the lower-shore ecotype. In contrast, only 4-8% of the upper-shore ecotype but 53% of large (> 6 mm) and 8% of small (3-6 mm) native lower-shore ecotype survived in the lower shores. In the midshores, both the two parental ecotypes and the hybrids survived about equally well. Thus there is a considerable advantage for the native ecotypes in the upper and lower shores, while in the hybrid zone none of the morphs, hybrids included, are favored. This indicates that the dimorphism of L. saxatilis is maintained by steep cross-shore selection gradients, thus supporting the selection-gradient model of hybrid zones. We performed field and laboratory experiments that suggest physical factors and predation as important selective agents. Earlier studies indicate assortative mating between the two ecotypes in the midshore. This is unexpected in a hybrid zone maintained by selection gradients, and it seems as if the reproductive barrier compresses the hybrid zone considerably.