Legacy effects of premature defoliation in response to an extreme drought event modulate phytochemical profiles with subtle consequences for leaf herbivory in European beech.

Extreme droughts can have long-lasting effects on forest community dynamics and species interactions. Yet, our understanding of how drought legacy modulates ecological relationships is just unfolding. We tested the hypothesis that leaf chemistry and herbivory show long-term responses to premature defoliation caused by an extreme drought event in European beech (Fagus sylvatica L.). For two consecutive years after the extreme European summer drought in 2018, we collected leaves from the upper and lower canopy of adjacently growing drought-stressed and unstressed trees. Leaf chemistry was analyzed and leaf damage by different herbivore-feeding guilds was quantified. We found that drought had lasting impacts on leaf nutrients and on specialized metabolomic profiles. However, drought did not affect the primary metabolome. Drought-related phytochemical changes affected damage of leaf-chewing herbivores whereas damage caused by other herbivore-feeding guilds was largely unaffected. Drought legacy effects on phytochemistry and herbivory were often weaker than between-year or between-canopy strata variability. Our findings suggest that a single extreme drought event bears the potential to long-lastingly affect tree-herbivore interactions. Drought legacy effects likely become more important in modulating tree-herbivore interactions since drought frequency and severity are projected to globally increase in the coming decades.

A glimmer of hope - ash genotypes with increased resistance to ash dieback pathogen show cross-resistance to emerald ash borer.

Plants rely on cross-resistance traits to defend against multiple, phylogenetically distinct enemies. These traits are often the result of long co-evolutionary histories. Biological invasions can force naïve plants to cope with novel, coincident pests, and pathogens. For example, European ash (Fraxinus excelsior) is substantially threatened by the emerald ash borer (EAB), Agrilus planipennis, a wood-boring beetle, and the ash dieback (ADB) pathogen, Hymenoscyphus fraxineus. Yet, plant cross-resistance traits against novel enemies are poorly explored and it is unknown whether naïve ash trees can defend against novel enemy complexes via cross-resistance mechanisms. To gain mechanistic insights, we quantified EAB performance on grafted replicates of ash genotypes varying in ADB resistance and characterized ash phloem chemistry with targeted and untargeted metabolomics. Emerald ash borer performed better on ADB-susceptible than on ADB-resistant genotypes. Moreover, changes in EAB performance aligned with differences in phloem chemical profiles between ADB-susceptible and ADB-resistant genotypes. We show that intraspecific variation in phloem chemistry in European ash can confer increased cross-resistance to invasive antagonists from different taxonomic kingdoms. Our study suggests that promotion of ADB-resistant ash genotypes may simultaneously help to control the ADB disease and reduce EAB-caused ash losses, which may be critical for the long-term stability of this keystone tree species.

Genotypic variation rather than ploidy level determines functional trait expression in a foundation tree species in the presence and absence of environmental stress.

BACKGROUND AND AIMS: At the population level, genetic diversity is a key determinant of a tree species' capacity to cope with stress. However, little is known about the relative importance of the different components of genetic diversity for tree stress responses. We compared how two sources of genetic diversity, genotype and cytotype (i.e. differences in ploidy levels), influence growth, phytochemical and physiological traits of Populus tremuloides in the presence and absence of environmental stress. METHODS: In a series of field studies, we first assessed variation in traits across diploid and triploid aspen genotypes from Utah and Wisconsin under non-stressed conditions. In two follow-up experiments, we exposed diploid and triploid aspen genotypes from Wisconsin to individual and interactive drought stress and defoliation treatments and quantified trait variations under stress. KEY RESULTS: We found that (1) tree growth and associated traits did not differ significantly between ploidy levels under non-stressed conditions. Instead, variation in tree growth and most other traits was driven by genotypic and population differences. (2) Genotypic differences were critical for explaining variation of most functional traits and their responses to stress. (3) Ploidy level played a subtle role in shaping traits and trait stress responses, as its influence was typically obscured by genotypic differences. (4) As an exception to the third conclusion, we showed that triploid trees expressed 17 % higher foliar defence (tremulacin) levels, 11 % higher photosynthesis levels and 23 % higher rubisco activity under well-watered conditions. Moreover, triploid trees displayed greater drought resilience than diploids as they produced 35 % more new tissue than diploids when recovering from drought stress. CONCLUSION: Although ploidy level can strongly influence the ecology of tree species, those effects may be relatively small in contrast to the effects of genotypic variation in highly diverse species.

Genetic divergence along a climate gradient shapes chemical plasticity of a foundation tree species to both changing climate and herbivore damage.

Climate change is threatening the persistence of many tree species via independent and interactive effects on abiotic and biotic conditions. In addition, changes in temperature, precipitation, and insect attacks can alter the traits of these trees, disrupting communities and ecosystems. For foundation species such as Populus, phytochemical traits are key mechanisms linking trees with their environment and are likely jointly determined by interactive effects of genetic divergence and variable environments throughout their geographic range. Using reciprocal Fremont cottonwood (Populus fremontii) common gardens along a steep climatic gradient, we explored how environment (garden climate and simulated herbivore damage) and genetics (tree provenance and genotype) affect both foliar chemical traits and the plasticity of these traits. We found that (1) Constitutive and plastic chemical responses to changes in garden climate and damage varied among defense compounds, structural compounds, and leaf nitrogen. (2) For both defense and structural compounds, plastic responses to different garden climates depended on the climate in which a population or genotype originated. Specifically, trees originating from cool provenances showed higher defense plasticity in response to climate changes than trees from warmer provenances. (3) Trees from cool provenances growing in cool garden conditions expressed the lowest constitutive defense levels but the strongest induced (plastic) defenses in response to damage. (4) The combination of hot garden conditions and simulated herbivory switched the strategy used by these genotypes, increasing constitutive defenses but erasing the capacity for induction after damage. Because Fremont cottonwood chemistry plays a major role in shaping riparian communities and ecosystems, the effects of changes in phytochemical traits can be wide reaching. As the southwestern US is confronted with warming temperatures and insect outbreaks, these results improve our capacity to predict ecosystem consequences of climate change and inform selection of tree genotypes for conservation and restoration purposes.

Constitutive and induced insect resistance in RNAi-mediated ultra-low gossypol cottonseed cotton.

BACKGROUND: Besides fibers, cotton plants also produce a large amount of seeds with a high oil and protein content. The use of these seeds is restricted by their high contents of the terpenoid gossypol, which is harmful to humans and livestock. Using a genetic engineering approach, "Ultra-low gossypol cottonseed" (ULGCS) plants were produced by knocking down an enzyme that catalyzes the formation of a precursor of gossypol. This was accomplished via RNAi-mediated silencing of the target gene using a seed-specific α-globulin promotor. Since gossypol is also a crucial defense mechanism against leaf-feeding herbivores, ULGCS plants might possess lower herbivore resistance than non-engineered plants. Therefore, we tested the constitutive and inducible direct insect resistance of two ULGCS cotton lines against the African cotton leafworm, Spodoptera littoralis. RESULT: The herbivore was equally affected by both ULGCS lines and the control (Coker 312) line when feeding on fully expanded true leaves from undamaged plants and plants induced by jasmonic acid. When plants were induced by caterpillar-damage, however, S. littoralis larvae performed better on the ULGCS plants. Terpenoid analyses revealed that the ULGCS lines were equally inducible as the control plants. Levels of terpenoids were always lower in one of the two lines. In the case of cotyledons, caterpillars performed better on ULGCS cotton than on conventional cotton. This was likely caused by reduced levels of gossypol in ULGCS cotyledons. CONCLUSION: Despite those effects, the insect resistance of ULGSC cotton can be considered as largely intact and the plants may, therefore, be an interesting alternative to conventional cotton varieties.

Differential Impact of Herbivores from Three Feeding Guilds on Systemic Secondary Metabolite Induction, Phytohormone Levels and Plant-Mediated Herbivore Interactions.

Phytochemical defense responses of plants are often herbivore-specific and can be affected by a herbivore's feeding mode. However, comprehensive studies documenting the impact of multiple herbivores from different feeding guilds on induced phytochemical responses in distal leaves and its consequences for plant-mediated herbivore interactions are limited and findings are inconsistent. We investigated how herbivory by leaf-chewing caterpillars, cell-content feeding spider mites and phloem-feeding aphids and whiteflies affect secondary metabolomes and phytohormone levels in youngest, non-damaged cotton leaves (distal leaves). Furthermore, bioassays with caterpillars were conducted to assess their performance on distal leaves of plants infested with different herbivores. Caterpillars, and to a lesser degree spider mites, led to a systemic induction of terpenoids with negative consequences for caterpillar performance in the bioassays. Both herbivores reduced levels of various nutrients and potentially antioxidative compounds. Caterpillar damage increased levels of jasmonoyl-L-isoleucine and abscisic acid (ABA), whereas spider mite infestation had no effect on phytohormone levels. Aphid and whitefly infestation did not systemically affect secondary metabolites. Aphids decreased salicylic acid levels while whitefly-infested plants contained increased ABA levels. Neither aphid nor whitefly infestation affected caterpillar performance. In general, feeding mode of a herbivore can affect systemically induced changes in phytochemistry and plant-mediated indirect interactions even though the two phloem-feeding herbivores triggered different phytohormonal responses. The observed reduction of nutrients and potentially antioxidative compounds upon caterpillar and spider mite herbivory underlines the importance of further elucidating the role of resource sequestration as a potential systemic defensive response following herbivory by chewers and cell-content feeding herbivores.

Transcriptome profiling of Fraxinus excelsior genotypes infested by emerald ash borer.

European ash, Fraxinus excelsior is facing the double threat of ongoing devastation by the invasive fungal pathogen, Hymenoscyphus fraxineus and the imminent arrival of the non-native emerald ash borer (EAB), Agrilus planipennis. The spread of EAB which is currently moving westwards from European Russia and Ukraine into central Europe, poses an additional substantial threat to European ash, F. excelsior. While the molecular basis for resistance or variation in resistance among European ash genotypes is heavily investigated, comparatively little is known about the molecular ash traits involved in resistance against EAB. In this study we have gathered transcriptomic data from EAB inoculated genotypes of F. excelsior that have previously shown different levels of susceptibility to EAB. Resultant datasets show differential gene expression in susceptible and resistant genotypes in response to EAB infestation. This data will provide important information on the molecular basis of resistance to the EAB and allow the development of management plans to combat a pending threat of a culturally and ecologically important European tree species.

Host searching and host preference of resident pupal parasitoids of Drosophila suzukii in the invaded regions.

BACKGROUND: In its invaded regions, Drosophila suzukii (Matsumura) is a novel host for the community of resident parasitoids of Drosophila. To attain a high parasitization rate on the novel host, the parasitoids have to locate it and accept it in the presence of other Drosophila hosts. We conducted a laboratory choice experiment and a semifield trial to investigate host searching and host preference of the three pupal parasitoid species Trichopria drosophilae (Perkins), Pachycrepoideus vindemmiae (Rondani) and Spalangia erythromera Förster. RESULTS: All three parasitoid species preferred D. suzukii over two common native hosts in the choice experiment. In field cages, most parasitoid offspring emerged from D. suzukii hosts. While P. vindemmiae mainly parasitized hosts in the foliage, most T. drosophilae offspring emerged from pupae presented on the ground. CONCLUSIONS: Both P. vindemmiae and T. drosophilae have the potential to find and parasitize D. suzukii in the field. If released early in the season, possible nontarget effects on native Drosophila should be minimal.

Reduced caterpillar damage can benefit plant bugs in Bt cotton.

Bt cotton was genetically modified to produce insecticidal proteins targeting Lepidopteran pests and is therefore only minimally affected by caterpillar damage. This could lead to reduced levels of inherent, systemically inducible defensive compounds in Bt cotton which might benefit other important cotton herbivores such as plant bugs. We studied the effects of plant defense induction on the performance of the plant bug Lygus hesperus by caging nymphs on different food sources (bolls/squares) of Bt and non-Bt cotton which were either undamaged, damaged by Bt tolerant caterpillars, or treated with jasmonic acid (JA). Terpenoid induction patterns of JA-treated and L. hesperus-damaged plants were characterized for different plant structures and artificial diet assays using purified terpenoids (gossypol/heliocide H1/4) were conducted. Nymphs were negatively affected if kept on plants damaged by caterpillars or sprayed with JA. Performance of nymphs was increased if they fed on squares and by the Bt-trait which had a positive effect on boll quality as food. In general, JA-sprayed plants (but not L. hesperus infested plants) showed increased levels of terpenoids in the plant structures analyzed, which was especially pronounced in Bt cotton. Nymphs were not negatively affected by terpenoids in artificial diet assays indicating that other inducible cotton responses are responsible for the found negative effects on L. hesperus. Overall, genetically engineered plant defenses can benefit plant bugs by releasing them from plant-mediated indirect competition with lepidopterans which might contribute to increasing numbers of hemipterans in Bt cotton.

Interaction of transgenic and natural insect resistance mechanisms against Spodoptera littoralis in cotton.

BACKGROUND: Insect-resistant transgenic plants that express insecticidal Cry proteins from Bacillus thuringiensis (Bt) are grown on millions of hectares worldwide. While these proteins are efficient in controlling key lepidopteran pests, not all pests are affected and the development of resistance in target pests is always a concern. These shortcomings could be addressed by exploiting the natural insect resistance of cotton, especially inducible terpenoids such as gossypol. RESULTS: To assess the potential of gossypol in supplementing Cry proteins as a resistance trait, we conducted a range of feeding assays with Spodoptera littoralis using artificial diet with defined amounts of Cry proteins and gossypol. This was supplemented by assays with leaf discs of induced and uninduced non-Bt and Bt cotton (expressing Cry1Ac and Cry2Ab). Additionally, we quantified Cry proteins and cotton terpenoids to describe the interactions in planta. We found that gossypol can increase the efficacy of Cry proteins in artificial diet in an additive way. Induced production of gossypol and other cotton terpenoids, however, did not increase the efficacy of Bt cotton owing to the strong impact of the Bt trait. CONCLUSION: Cotton terpenoids may offer the chance to supplement the insect resistance of Bt cotton in cases were the pest is not strongly affected by the Cry proteins. © 2016 Society of Chemical Industry.

Cotton Defense Induction Patterns Under Spatially, Temporally and Quantitatively Varying Herbivory Levels.

In its defense against herbivores, cotton (Gossypium sp.) relies in part on the production of a set of inducible, non-volatile terpenoids. Under uniform damage levels, in planta allocation of induced cotton terpenoids has been found to be highest in youngest leaves, supporting assumptions of the optimal defense theory (ODT) which predicts that plants allocate defense compounds to tissues depending on their value and the likelihood of herbivore attack. However, our knowledge is limited on how varying, and thus more realistic, damage levels might affect cotton defense organization. We hypothesized that the allocation of terpenoids and densities of terpenoid-storing glands in leaves aligns with assumptions of the ODT, even when plants are subjected to temporally, spatially and quantitatively varying caterpillar (Heliothis virescens) damage. As expected, cotton plants allocated most of their defenses to their youngest leaves regardless of damage location. However, defense induction in older leaves varied with damage location. For at least 14 days after damage treatments ended, plants reallocated defense resources from previously young leaves to newly developed leaves. Furthermore, we observed a positive hyperbolic relationship between leaf damage area and both terpenoid concentrations and gland densities, indicating that cotton plants can fine-tune defense allocation. Although it appears that factors like vascular constraints and chemical properties of individual defense compounds can affect defense levels, our results overall demonstrate that induced defense organization of cotton subjected to varying damage treatments is in alignment with key assumptions of the ODT.