Genomic dissection of an extended phenotype: Oak galling by a cynipid gall wasp.

Galls are plant tissues whose development is induced by another organism for the inducer's benefit. 30,000 arthropod species induce galls, and in most cases the inducing effectors and target plant systems are unknown. Cynipid gall wasps are a speciose monophyletic radiation that induce structurally complex galls on oaks and other plants. We used a model system comprising the gall wasp Biorhiza pallida and the oak Quercus robur to characterise inducer and host plant gene expression at defined stages through the development of galled and ungalled plant tissues, and tested alternative hypotheses for the origin and type of galling effectors and plant metabolic pathways involved. Oak gene expression patterns diverged markedly during development of galled and normal buds. Young galls showed elevated expression of oak genes similar to legume root nodule Nod factor-induced early nodulin (ENOD) genes and developmental parallels with oak buds. In contrast, mature galls showed substantially different patterns of gene expression to mature leaves. While most oak transcripts could be functionally annotated, many gall wasp transcripts of interest were novel. We found no evidence in the gall wasp for involvement of third-party symbionts in gall induction, for effector delivery using virus-like-particles, or for gallwasp expression of genes coding for plant hormones. Many differentially and highly expressed genes in young larvae encoded secretory peptides, which we hypothesise are effector proteins exported to plant tissues. Specifically, we propose that host arabinogalactan proteins and gall wasp chitinases interact in young galls to generate a somatic embryogenesis-like process in oak tissues surrounding the gall wasp larvae. Gall wasp larvae also expressed genes encoding multiple plant cell wall degrading enzymes (PCWDEs). These have functional orthologues in other gall inducing cynipids but not in figitid parasitoid sister groups, suggesting that they may be evolutionary innovations associated with cynipid gall induction.

UCE data reveal multiple origins of rose gallers in North America: Global phylogeny of Diplolepis Geoffroy (Hymenoptera: Cynipidae).

Gall wasps in the genus Diplolepis Geoffroy are specialized herbivores that induce galls exclusively on roses. Despite their wide distribution across the Holarctic, little is known about their evolutionary history. Here we present the first phylogenomic tree of global Diplolepis reconstructed using Ultraconserved Elements (UCEs), resulting in a robust phylogeny based on 757 genes. Results support the existence of two principal clades: a Nearctic stem-galler clade, and a Holarctic leaf-galler clade that further splits into two Palearctic groups and one Nearctic group. This topology is congruent with a previous study based on the mitochondrial gene COI, an unexpected result given the common occurrence of mitonuclear discordance in closely related oak gall wasp lineages. Most Diplolepis species were recovered as reciprocally monophyletic, with some notable exceptions such as the D. polita and the D. ignota complex, for which species boundaries remain unresolved. Historical biogeographic reconstruction was unable to pinpoint the origin of Diplolepis, but confirms two independent incursions into the Nearctic. Ancestral state reconstruction analysis highlights the conservatism of gall location on the host plants, as shifts to different host organs are relatively rare. We suggest that Diplolepis were originally leaf gallers, with a Nearctic stem-galler clade undergoing a major plant organ switch onto rose stems. Host organ switch or reversal is uncommon, which suggests a level of conservatism. Our study showcases the resolving power of UCEs at the species level while also suggesting improvements to advance future Cynipoidea phylogenomics. Our results also highlight the additional sampling needed to clarify taxonomic relationships in the Nearctic and eastern Palearctic regions.

Deleterious effects of mild simulated overwintering temperatures on survival and potential fecundity of rose-galling Diplolepis wasps (Hymenoptera: Cynipidae).

Prepupae of the rose galling Diplolepis spinosa from areas with relatively cold winters in southern Canada, and Diplolepis variabilis from a milder locale in western Canada, were used to test the hypothesis that mild winter temperatures are detrimental to the survival and potential fecundity of insects. Prepupae of D. spinosa held within or removed from their galls were exposed to simulated overwintering temperatures (-22, 0, 5, or 10 degrees C) for approximately four months before measuring their survival, body size, and potential fecundity. Similar studies were conducted using prepupae of D. variabilis that were removed from their gall and subjected to 0 degrees C or 10 degrees C treatments. Diplolepis spinosa, with or without their galls, averaged 66% more mortality at 10 degrees C than at 0 degrees C. Female D. spinosa that survived the 10 degrees C treatment had 32% fewer eggs than those held at 0 degrees C. In contrast, there was no difference in survival or numbers of eggs between D. variabilis held at 0 degrees C and 10 degrees C. Body size of adult females and size of eggs did not differ among temperature treatments for either species. We conclude that mild overwintering temperatures may be detrimental for insects by raising their metabolism, and consequently reducing energetic reserves needed for development to the adult stage and subsequent production of eggs the following spring.

Extreme resistance to desiccation and microclimate-related differences in cold-hardiness of gall wasps (Hymenoptera: Cynipidae) overwintering on roses in southern Canada.

Four species of cynipid wasp of the genus Diplolepis that induce galls on roses (Rosa species) in southern Canada and two species of inquiline cynipid associated with these galls were studied for their cold-hardiness and resistance to water loss and for possible links between these adaptations. Mid-winter-acclimated supranivean D. spinosa and Periclistus pirata had lower supercooling points (-38 to -40 degrees C) and higher hemolymph osmolalities (1760-1849 mosmol kg(-1)) than subnivean D. polita, D. gracilis, D. radicum and Periclistus sp. (-31 to -32 degrees C and 977-1464 mosmol kg(-1), respectively). During a simulated transition from summer/fall to mid-winter conditions, the glycerol concentration of D. spinosa more than tripled, reaching a final value of 0.98 moll(-1), while its supercooling point decreased by 13 degrees C from the initial value of -27.4 degrees C; however, glycerol concentration and supercooling point did not change for the subnivean species. The permeability of the cuticle of all species was extremely low (0.33-1.00 microg h(-1) cm(-2) mmHg(-1) at 5 degrees C and 0 % relative humidity; 1 mmHg=0.133 kPa), even compared with that of desert species; however, there was no difference in cuticular permeability between supranivean and subnivean prepupae. Transition temperatures ranged between 32.3 and 34.6 degrees C; below 30 degrees C, temperature had little effect on rates of water loss for all species (Q(10)=1.13-1.87).