Insect egg-induced physiological changes and transcriptional reprogramming leading to gall formation.

Gall-inducing insects and their hosts present some of the most intricate plant-herbivore interactions. Oviposition on the host is often the first cue of future herbivory and events at this early time point can affect later life stages. Many gallers are devastating plant pests, yet little information regarding the plant-insect molecular interplay exists, particularly following egg deposition. We studied the physiological and transcriptional responses of Eucalyptus following oviposition by the gall-inducing wasp, Leptocybe invasa, to explore potential mechanisms governing defence responses and gall development. RNA sequencing and microscopy were used to explore a susceptible Eucalyptus-L. invasa interaction. Infested and control material was compared over time (1-3, 7 and 90 days post oviposition) to examine the transcriptional and morphological changes. Oviposition induces accumulation of reactive oxygen species and phenolics which is reflected in the transcriptome analysis. Gene expression supports phytohormones and 10 transcription factor subfamilies as key regulators. The egg and oviposition fluid stimulate cell division resulting in gall development. Eucalyptus responses to oviposition are apparent within 24 hr. Putative defences include the oxidative burst and barrier reinforcement. However, egg and oviposition fluid stimuli may redirect these responses towards gall development.

Performance of the thistle gall fly, Urophora cardui, in relation to host plant nitrogen and mycorrhizal colonization.

Many hypotheses have been developed to explain the adaptive nature of insect galls. One of these, the nutrition hypothesis, states that gall formers have advantages over other insects because gall tissue provides a better (higher quality) food source than unmodified tissue. However, this has rarely been experimentally tested. In a test of this hypothesis, we grew plants of Cirsium arvense (L.) Scop. in a factorial design with two main treatments: the addition of nitrogen (to enhance foliar N levels) and of fungicide (to reduce colonization of roots by arbuscular mycorrhizal fungi). Mycorrhizal fungi have been shown previously to reduce the N concentration of host plants. Plants were exposed to adult gall flies, Urophora cardui L., and maintained through one season to allow maturation of galls. Reduction of the percentage mycorrhizal colonization by fungicide resulted in an elevation of total stem N comparable to that achieved by N addition, but gall N concentration remained unchanged in all treatments. Nitrogen application elevated stem N levels when mycorrhizal fungi were present, but application of both compounds together did not result in any increase over either single treatment. Fungicide application resulted in larger galls, which contained more larval chambers, with more live, and heavier, larvae. However, the main effects of N were not significant, as N addition only increased fly performance on plants where mycorrhizas were not reduced. It is suggested that U. cardui gall inhabitants can manipulate N at an optimal level and thus might conform to a modified version of the nutrition hypothesis. Mycorrhizal colonization might reduce gall fly performance by delaying the appearance, or impairing the quality, of secondary nutritive tissue in the gall. Future tests of the nutrition hypothesis should include a consideration of the plant's mycorrhizal status.

Constraints on growth and allocation patterns of Silphium integrifolium (Asteraceae) caused by a cynipid gall wasp.

Insect herbivory can have important effects on plant life histories and architecture. We quantified the impact that a cynipid gall wasp, Antistrophus silphii, had on growth, reproduction, and biomass allocation patterns of Silphium integrifolium growing in the tallgrass prairie of northeastern Kansas. Experimentally galled individual Silphium shoots (ramets) had reduced shoot growth, leaf and flower head production, and delayed flowering compared to gall-free control shoots. Gall formation completely halted normal apical growth in 65% of the shoots. Galling did not affect individual flower head weight, the numbers of achenes per flower head or achene weight. Silphium plants (genets) with a high proportion of galled shoots had lower total biomass, a lower proportion of total biomass allocated to flower heads, higher allocation to leaves, but no change in allocation to stems or rhizome. High gall densities reduced the number of flower heads per plant and shortened the time between flower head initiation and maturity. An adaptive interpretation of these results would be that the survivorship and future performance of galled Silphium may be promoted by maintaining allocation to rhizome. However, reduced shoot growth and delayed reproduction in galled Silphium may weaken its competitive ability and reduce pollination success, so that any adaptive advantage to Silphium's allocation responses to galls may be outweighed by disadvantages from its growth and flowering phenology responses. We conclude that a more parsimonious interpretation of these results is that gall-induced allocation changes are due to architectural constraints placed by galls on meristem activity, rather than to any adaptive response on the part of the plant.

Goldenrod stem galler preference and performance: effects of multiple herbivores and plant genotypes.

Few studies have examined how the preference-performance relationship of an herbivore for different genotypes of its host plant is affected by the presence and/or feeding activity of other members of the herbivore assemblage. In an outdoor garden, we manipulated the abundance of three common herbivores, the meadow spittlebug, a leaf beetle, and an aphid, on replicate 1-m2 plots of 16 different genotypes of tall goldenrod, Solidago altissima. Adults of the goldenrod stem galler, Eurosta solidaginis, were subsequently released into the garden to oviposit among the host plants. Oviposition preference was strongly influenced by plant genotype and the presence of two of the herbivores, spittlebugs and leaf beetles. The effects of the herbivores were additive: the presence of leaf beetles reduced preference by 6%, spittlebugs by 18%, and both herbivores combined by 25%. Plant genotype-herbivore species interaction effects on stem-galler preference, which would indicate the presence of genetic variation among goldenrod genotypes in their norms of reaction for their acceptability as a host to the stem galler, were absent in this study. The performance of the stem galler was also significantly affected by goldenrod genotype, but in general was not affected by the presence of herbivores early in the season (the exception was a positive correlation between the proportion of ramets infested by all herbivores and gall size). Overall, we could find no correlation between preference and performance. This is in accord with results from previous studies on this system that were performed in the absence of herbivores, suggesting that the presence of herbivores in this study did not qualitatively alter the preference-performance relationship. We suggest that the lack of a positive correlation between host-plant preference and larval performance may reflect a constraint on the discriminatory ability of female stem gallers preventing them from selecting the best hosts among plants that differ in genotype and level of environmental stress (e.g., presence of interspecific herbivores).

The functional resource of a gall-forming adelgid.

Interactions among shoots within plant modules could allow gall-insects to acquire resources from other plant parts near the feeding sites. As a result, nearby plant parts may act as a "functional resource", or extended resource base. We tested for functional interconections between galls and adjacent ungalled shoots in Adelges cooleyi Gil. (Homoptera: Adelgidae) on Picea engelmanni, Engelmann spruce. Observations of gall and surrounding shoot weights showed that gall weights were twice normal shoot weights, but that surrounding shoot weights were unaffected. Reducing photosynthate availability by covering galls or surrounding shoots with opaque cloth suggested that functional interconnections exist among them; covering galls reduced surrounding current-year ungalled shoot weight, and covering surrounding shoots reduced their weight even further, but neither covering treatment affected gall or gall-insect weight. These patterns suggest that surrounding shoots constitute an extended and flexibly utilized resource base for adelgid galls. Resources made available via functional interconnections appeared to be equally available throughout adelgid galls. No differences were found in gall-insect performance in parts of the gall closer to connections with surrounding shoots compared to more distantly-located gall-insects. Further studies are required to examine patterns of resource movement among unmanipulated galls and shoots. Functional resources may be common features of plant/gall-insect interactions, potentially playing an important role in gall-insect reproductive success and habitat selection.

Increased photosynthesis and water potentials in Silphium integrifolium galled by cynipid wasps.

Interactions between drought, insect herbivory, photosynthesis, and water potential play a key role in determining how plants tolerate and defend against herbivory, yet the effects of insect herbivores on photosynthesis and water potential are seldom assessed. We present evidence that cynipid wasp galls formed by Antistrophus silphii on Silphium integrifolium increase photosynthesis (A), stomatal conductance (g), and xylem water potential (Ψ). Preliminary data showed that in drought-stressed plants galled shoots had 36% greater A, and 10% greater stem Ψ than ungalled shoots, while in well-watered plants leaf gas exchange was not affected by galls. We hypothesized that 1) galled shoots have higher Ψ, g, and A than ungalled shoots, but this differences diminishes if plant drought stress is reduced, and 2) galls can reduce decreases in A and g if water availability decreases. A field experiment testing the first hypothesis found that galls increased g and Ψ, but that differences between galled and ungalled shoots did not diminish after plants were heavily watered. A laboratory test of the second hypothesis using potted Silphium found that galled plants had smaller drops in A and g over a 4-day dry-down period. A vs g and A vs intercellular CO2 concentration relationships were consistent with the explanation that increased Ψ allows galls to increase A by reducing stomatal limitation of A, rather than by altering sink-source relationships or by removing low-Ψ limitations on non-stomatal components of A. Our working hypothesis is that galls increase Ψ and A by reducing the shoot: root ratio so that the plant is exploiting a greater soil volume per unit leaf area. We argue that increased A is an ineffective way for Silphium to compensate for negative effects of gall insect attack. Instead, increased Ψ and A may protect gall insects from variation in resource availability caused by periodic drought stress, potentially reducing negative effects of drought on plant quality and on gall insect populations.