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.

A closer look at the spatial architecture of aphid clones.

Nearly 25 years ago, Ellstrand & Roose (1987) reviewed what was known at the time of the genetic structure of clonal plant species. What is the relationship between space and clonal fitness, they asked. What is the best way for a clone to grow within its ecological neighbourhood? The pot had been stirred 10 years previously by Janzen (1977), who pointed out how little we know about the population biology of clonal organisms like dandelions and aphids. He wondered whether, like good curries, outward appearances masked common ingredients. Because in no small part of the advent of molecular ecology, we know more about clonal life histories today, particularly in plants (van Dijk 2003; Vallejo-Marin et al. 2010). Surprisingly, studies of the spatial architecture of aphid clones have been comparably rare. In this issue of Molecular Ecology, Vantaux et al. characterize the finescale distribution of the black bean aphid (Aphis fabae) and in so doing, help to fill that gap. They describe a moderate degree of intermingling between aphid clones over a growing season--A. fabae clones are 'sticky', but only a bit. By mixing, clones directly compete with each other as well. The results of Vantaux et al. (2011) will help to integrate evolutionary patterns in aphids with the appropriate ecological scales out of which those patterns emerge.

Symbiont-mediated phenotypic variation without co-evolution in an insect-fungus association.

Recent studies have shown that symbionts can be a source of adaptive phenotypic variation for their hosts. It is assumed that co-evolution between hosts and symbionts underlies these ecologically significant phenotypic traits. We tested this assumption in the ectosymbiotic fungal associate of the gall midge Asteromyia carbonifera. Phylogenetic analysis placed the fungal symbiont within a monophyletic clade formed by Botryosphaeria dothidea, a typically free-living (i.e. not associated with an insect host) plant pathogen. Symbiont isolates from four divergent midge lineages demonstrated none of the patterns common to heritable microbial symbioses, including parallel diversification with their hosts, substitution rate acceleration, or A+T nucleotide bias. Amplified fragment length polymorphism genotyping of the symbiont revealed that within-lineage genetic diversity was not clustered along host population lines. Culture-based experiments demonstrated that the symbiont-mediated variation in gall phenotype is not borne out in the absence of the midge. This study shows that symbionts can be important players in phenotypic variation for their hosts, even in the absence of a co-evolutionary association.

The cost of conflict in aphid societies.

Most social aphids are found within plant galls, inside of which clonally-derived family groups feed, and specialized larval castes forego reproduction and perform various cooperative tasks, including group defence. When unrelated aphids move between clones, conditions are ripe for conflict because galls and cooperative defence are shared resources that are vulnerable to exploitation. A key unknown is whether conflict is costly in aphid social groups. We show that diversity within groups is negatively correlated with performance in the North American social aphid, Pemphigus obesinymphae. A substantial fraction of productivity is invested into drifting. However, drifting aphids tend to mature and depart non-natal galls prior to the seasonal peak in fecundity. These results suggest that when unrelated individuals move between groups, social aphids may experience conditions consistent with a tragedy of the commons. These results also emphasize the strongly convergent properties associated with conflict across the spectrum of animal and microbial sociality.

Extremely low levels of genetic polymorphism in endosymbionts (Buchnera) of aphids (Pemphigus).

Molecular evolutionary studies have suggested that vertically transmitted endosymbionts are subject to accumulation of deleterious mutations through genetic drift. Predictions of this hypothesis for patterns of intraspecific polymorphism were borne out in the single relevant study available, on the symbiont Buchnera aphidicola of Uroleucon ambrosiae. In order to examine the generality of this result, we surveyed DNA sequence variation in Buchnera of the distantly related aphid, Pemphigus obesinymphae. In contrast to Uroleucon species, Pemphigus species have complex life cycles with few dispersal stages. Despite these differences, P. obesinymphae showed patterns of variation at two Buchnera loci and one mitochondrial locus that were remarkably similar to those reported previously for Buchnera of U. ambrosiae. In the western US, Buchnera was nearly monomorphic, and in the eastern US, synonymous divergence ranged from 0.08 to 0.16%. Most polymorphisms involved rare alleles, consistent with a recent range of ancestral polymorphism, probably due to demographic fluctuations in aphid populations. These results support the generality of small effective population size in Buchnera and their aphid hosts.

Genetic conflict and conditional altruism in social aphid colonies.

Although kin selection is central to the modern study of social evolution, recent studies of social species have revealed that no simple relationship exists between levels of kinship and sociality. The soldier-producing aphids are unique among highly social animals because, barring movement by aphids between colonies, they occur in clonal groups of genetically identical individuals. Potentially, clonality simplifies efforts to understand social evolution in aphids by obviating issues of intragroup conflict. However, we report here high levels of clonal mixing and conflict in an aphid society. The gall-dwelling colonies of a social aphid species (Pemphigus obesinymphae) are not pure clones, but are invaded by large numbers of aphids from other clones. Intruders behave and develop selfishly once they have invaded a colony of nonkin. They refrain from risky defensive behaviors and accelerate their own development into reproductive rather than defensive stages. This conditionality in the social life of P. obesinymphae reveals complex dynamics and a degree of behavioral plasticity not previously known in aphid societies.

Individual and population variation in invertebrates revealed by Inter-simple Sequence Repeats (ISSRs).

PCR-based molecular markers are well suited for questions requiring large scale surveys of plant and animal populations. Inter-simple Sequence Repeats or ISSRs are analyzed by a recently developed technique based on the amplification of the regions between inverse-oriented microsatellite loci with oligonucleotides anchored in microsatellites themselves. ISSRs have shown much promise for the study of the population biology of plants, but have not yet been explored for similar studies of animals. The value of ISSRs is demonstrated for the study of animal species with low levels of within-population variation. Sets of primers are identified which reveal variation in two aphid species, Acyrthosiphon pisum and Pemphigus obesinymphae, in the yellow fever mosquito Aedes aegypti, and in a rotifer in the genus Philodina.

Cospeciation of psyllids and their primary prokaryotic endosymbionts.

Psyllids are plant sap-feeding insects that harbor prokaryotic endosymbionts in specialized cells within the body cavity. Four-kilobase DNA fragments containing 16S and 23S ribosomal DNA (rDNA) were amplified from the primary (P) endosymbiont of 32 species of psyllids representing three psyllid families and eight subfamilies. In addition, 0.54-kb fragments of the psyllid nuclear gene wingless were also amplified from 26 species. Phylogenetic trees derived from 16S-23S rDNA and from the host wingless gene are very similar, and tests of compatibility of the data sets show no significant conflict between host and endosymbiont phylogenies. This result is consistent with a single infection of a shared psyllid ancestor and subsequent cospeciation of the host and the endosymbiont. In addition, the phylogenies based on DNA sequences generally agreed with psyllid taxonomy based on morphology. The 3' end of the 16S rDNA of the P endosymbionts differs from that of other members of the domain Bacteria in the lack of a sequence complementary to the mRNA ribosome binding site. The rate of sequence change in the 16S-23S rDNA of the psyllid P endosymbiont was considerably higher than that of other bacteria, including other fast-evolving insect endosymbionts. The lineage consisting of the P endosymbionts of psyllids was given the designation Candidatus Carsonella (gen. nov.) with a single species, Candidatus Carsonella ruddii (sp. nov.).