Tree diversity enhances predation by birds but not by arthropods across climate gradients.

Tree diversity can promote both predator abundance and diversity. However, whether this translates into increased predation and top-down control of herbivores across predator taxonomic groups and contrasting environmental conditions remains unresolved. We used a global network of tree diversity experiments (TreeDivNet) spread across three continents and three biomes to test the effects of tree species richness on predation across varying climatic conditions of temperature and precipitation. We recorded bird and arthropod predation attempts on plasticine caterpillars in monocultures and tree species mixtures. Both tree species richness and temperature increased predation by birds but not by arthropods. Furthermore, the effects of tree species richness on predation were consistent across the studied climatic gradient. Our findings provide evidence that tree diversity strengthens top-down control of insect herbivores by birds, underscoring the need to implement conservation strategies that safeguard tree diversity to sustain ecosystem services provided by natural enemies in forests.

Temporal variation in tree diversity effects on birds and its implications for top-down control of insect herbivores in a tropical system.

Tree diversity promotes predator abundance and diversity, but evidence linking these effects to increased predation pressure on herbivores remains limited. In addition, tree diversity effects on predators can vary temporally as a function of environmental variation, or due to contrasting responses by different predator types. In a multi-year study, we assessed temporal variation in tree diversity effects on bird community abundance, diversity, and predation rates as a whole and by functional group based on feeding guild (omnivores vs. insectivores) and migratory status (migrant vs. resident). To this end, we conducted bird point counts in tree monocultures and polycultures and assessed attacks on clay caterpillars four times over a 2-year period in a tree diversity experiment in Yucatan, Mexico. Tree diversity effects on the bird community varied across surveys, with positive effects on bird abundance and diversity in most but not all surveys. Tree diversity had stronger and more consistent effects on omnivorous and resident birds than on insectivorous and migratory species. Tree diversity effects on attack rates also varied temporally but patterns did not align with variation in bird abundance or diversity. Thus, while we found support for predicted increases in bird abundance, diversity, and predation pressure with tree diversity, these responses exhibited substantial variation over time and the former two were uncoupled from patterns of predation pressure, as well as contingent on bird functional traits. These results underscore the need for long-term studies measuring responses by different predator functional groups to better understand tree diversity effects on top-down control.

Volatile-Mediated Signalling Between Potato Plants in Response to Insect Herbivory is not Contingent on Soil Nutrients.

Plant-plant signalling via volatile organic compounds (VOCs) has been studied intensively, but its contingency on abiotic conditions (e.g., soil nutrients, drought, warming) is poorly understood. To address this gap, we carried out a greenhouse experiment testing whether soil nutrients influenced signalling between potato (Solanum tuberosum) plants in response to insect leaf herbivory by the generalist caterpillar Spodoptera exigua. We placed pairs of plants in plastic cages, where one plant acted as a VOC emitter and the other as a receiver. We factorially manipulated soil nutrients for both emitter and receiver plants, namely: unfertilized (baseline soil nutrients) vs. fertilized (augmented nutrients). Then, to test for signalling effects, half of the emitters within each fertilization level were damaged by S. exigua larvae and the other half remained undamaged. Three days after placing larvae, we collected VOCs from emitter plants to test for herbivory and fertilization effects on VOC emissions and placed S. exigua larvae on receivers to test for signalling effects on leaf consumption and larval mass gain as proxies of induced resistance. We found that herbivory increased total VOC emissions and altered VOC composition by emitter plants, but these effects were not contingent on fertilization. In addition, bioassay results showed that receivers exposed to VOCs from herbivore-damaged emitters had lower levels of herbivory compared to receivers exposed to undamaged emitters. However, and consistent with VOC results, fertilization did not influence herbivore-induced signalling effects on receiver resistance to herbivory. In sum, we found evidence of S. exigua-induced signalling effects on resistance to herbivory in potato plants but such effects were not affected by increased soil nutrients. These results call for further work testing signalling effects under broader range of nutrient concentration levels (including nutrient limitation), teasing apart the effects of specific nutrients, and incorporating other abiotic factors likely to interact or covary with soil nutrients.

Effect of herbivore load on VOC-mediated plant communication in potato.

MAIN CONCLUSION: VOC emissions increased with herbivore load, but this did not result in concomitant increases in resistance in neighbouring plants, suggesting that communication occurred independently of herbivore load in emitter plants. Herbivore-damaged plants emit volatile organic compounds (VOCs) that can alert neighbours and boost their resistance. While VOC-mediated plant communication has been shown to be herbivore-specific, we know little about its contingency on variation in herbivore load. To address this knowledge gap, we tested herbivore load effects on VOC-mediated communication between potato plants (Solanum tuberosum) using the generalist herbivore Spodoptera exigua. First, we tested whether herbivore load (three levels: undamaged control, low, and high load) affected total VOC emissions and composition. Second, we matched emitter and receiver plants and subjected emitters to the same herbivore load treatments. Finally, we performed a bioassay with S. exigua on receivers to test for induced resistance due to VOC-mediated communication. We found that herbivory significantly increased total VOC emissions relative to control plants, and that such increase was greater under high herbivore load. In contrast, we found no detectable effect of herbivory, regardless of the load, on VOC composition. The communication experiment showed that VOCs released by herbivore-induced emitters boosted resistance in receivers (i.e., lower leaf damage than receivers exposed to VOCs released by control emitters), but the magnitude of such effect was similar for both levels of emitter herbivore load. These findings suggest that changes in VOCs due to variation in herbivore load do not modify the outcomes of plant communication.

Plant genetic relatedness and volatile-mediated signalling between Solanum tuberosum plants in response to herbivory by Spodoptera exigua.

It has been proposed that plant-plant signalling via herbivore-induced volatile organic compounds (VOCs) should be stronger between closely related than unrelated plants. However, empirical tests remain limited and few studies have provided detailed assessments of induced changes in VOCs emissions across plant genotypes to explain genetic relatedness effects. In this study, we tested whether airborne signalling in response to herbivory between Solanum tuberosum (potato) plants was contingent on plant genetic relatedness, and further investigated genotypic variation in VOCs potentially underlying signalling and its contingency on relatedness. We carried out a greenhouse experiment using 15 S. tuberosum varieties placing pairs of plants in plastic cages, i.e. an emitter and a receiver, where both plants were of the same genotype or different genotype thereby testing for self-recognition, an elemental form genetic relatedness effects. Then, for half of the cages within each level of relatedness the emitter plant was damaged by Spodoptera exigua larvae whereas for the other half the emitter was not damaged. Three days later, we placed S. exigua larvae on receivers to test for emitter VOC effects on leaf consumption and larval weight gain (i.e. induced resistance). In addition, we used a second group of plants subjected to the same induction treatment with the same S. tuberosum varieties to test for herbivore-induced changes in VOC emissions and variation in VOC emissions among these plant genotypes. We found that herbivory drove changes in VOC composition but not total emissions, and also observed quantitative and qualitative variation in constitutive and induced VOC emissions among varieties. Results from the bioassay showed that the amount of leaf area consumed and larval weight gain on receiver plants exposed to damaged emitters were significantly lower compared to mean values on receivers exposed to control emitters. However, and despite genotypic variation in induced VOCs, this signalling effect was not contingent on plant genetic relatedness. These findings provide evidence of VOCs-mediated signalling between S. tuberosum plants in response to S. exigua damage, but no evidence of self-recognition effects in signalling contingent on variation in VOC emissions among S. tuberosum varieties.

Effects of latitude and conspecific plant density on insect leaf herbivory in oak saplings and seedlings.

PREMISE: Abiotic factors and plant species traits have been shown to drive latitudinal gradients in herbivory, and yet, population-level factors have been largely overlooked within this context. One such factor is plant density, which may influence the strength of herbivory and may vary with latitude. METHODS: We measured insect herbivory and conspecific plant density (CPD) of oak (Quercus robur) seedlings and saplings along a 17° latitudinal gradient (2700 km) to test whether herbivory exhibited a latitudinal gradient, whether herbivory was associated with CPD, and whether such an association changed with latitude. RESULTS: We found a positive but saturating association between latitude and leaf herbivory. Furthermore, we found no significant relationship between CPD and herbivory, and such lack of density effects remained consistent throughout the sampled latitudinal gradient. CONCLUSIONS: Despite the apparently negligible influence of plant density on herbivory for Q. robur, further research with other plant taxa and in different types of plant communities are needed to investigate density-dependent processes shaping geographical variation in plant-herbivore interactions.

Apparent inhibition of induced plant volatiles by a fungal pathogen prevents airborne communication between potato plants.

Plant communication in response to insect herbivory has been increasingly studied, whereas that involving pathogen attack has received much less attention. We tested for communication between potato (Solanum tuberosum) plants in response to leaf infection by the fungal pathogen Sclerotinia sclerotiorum. To this end, we measured the total amount and composition of volatile organic compounds (VOCs) produced by control and infected emitter plants, as well as tested for induced resistance of receiver plants exposed to VOCs from emitters. We further tested for changes in the expression of defensive genes due to pathogen infection. Fungal infection did not significantly affect the total amount or composition of VOCs produced by emitter plants. Correspondingly, we found no evidence of higher resistance to the pathogen in receiver plants exposed to VOCs from infected emitters relative to control emitters. Molecular analyses indicated that pathogen infection drove a down-regulation of genes coding for VOC precursors, potentially explaining the absence of pathogen effects on VOC emissions and thus of communication. Overall, these results indicate no evidence of airborne communication between potato plants in response to fungal infection and point at pathogen inhibition of VOC emissions as a likely explanation for this result.

Micro-climatic effects on plant phenolics at the community level in a Mediterranean savanna.

Research has shown that warming and drought change plant phenolics. However, much of this work has centered on the effects of individual abiotic stressors on single plant species rather than the concurrent effects of multiple stressors at the plant community level. To address this gap, we manipulated rainfall and air temperature to test for their individual and interactive effects on the expression of leaf phenolics at the community level for annual plant species occurring in two habitat types (under oak tree canopies or in open grasslands) in a Mediterranean savanna. We found that augmented temperature had a significant positive effect on the community-weighted mean of total phenolics whereas reduced rainfall had no effect. In addition, we found no evidence of interactive effects between climatic stressors and these patterns remained consistent across habitat types. Overall, this study points at increasing efforts to investigate the linkages between climate change and community-level shifts in plant secondary chemistry.

Climate drives intraspecific differentiation in the expression of growth-defence trade-offs in a long-lived pine species.

Intraspecific variation in plant defences is expected to be the result of adaptive and plastic responses to environmental conditions, where trade-offs between growth and defences are thought to play a key role shaping phenotypic patterns in defensive investment. Axial resin ducts are costly defensive structures that remain imprinted in the tree rings of conifers, therefore being a valuable proxy of defensive investment along the trees' lifespan. We aimed to disentangle climate-driven adaptive clines and plastic responses to both spatial and temporal environmental variation in resin duct production, and to explore growth-defence trade-offs. To that aim, we applied dendrochronological procedures to quantify annual growth and resin duct production during a 31-year-period in a Mediterranean pine species, including trees from nine populations planted in two common gardens. Both genetic factors and plastic responses modulated annual resin duct production. However, we found no evidence of adaptive clines with climate gradients driving population differentiation. Our results revealed a marked physiological trade-off between growth and defences, where the slope of the trade-off was genetically variable and associated with climatic gradients. Our results help to enlighten the evolutionary patterns and genetic basis of defensive allocation within species, particularly revealing a key role of growth-defence trade-offs.

Resin ducts as resistance traits in conifers: linking dendrochronology and resin-based defences.

Conifers have evolved different chemical and anatomical defences against a wide range of antagonists. Resin ducts produce, store and translocate oleoresin, a complex terpenoid mixture that acts as both a physical and a chemical defence. Although resin duct characteristics (e.g., number, density, area) have been positively related to biotic resistance in several conifer species, the literature reporting this association remains inconclusive. Axial resin ducts recorded in annual growth rings are an archive of annual defensive investment in trees. This whole-life record of defence investment can be analysed using standard dendrochronological procedures, which allows us to assess interannual variability and the effect of understudied drivers of phenotypic variation on resin-based defences. Understanding the sources of phenotypic variation in defences, such as genetic differentiation and environmental plasticity, is essential for assessing the adaptive potential of forest tree populations to resist pests under climate change. Here, we reviewed the evidence supporting the importance of resin ducts in conifer resistance, and summarized current knowledge about the sources of variation in resin duct production. We propose a standardized methodology to measure resin duct production by means of dendrochronological procedures. This approach will illuminate the roles of resin ducts in tree defence across species, while helping to fill pivotal knowledge gaps in plant defence theory, and leading to a robust understanding of the patterns of variation in resin-based defences throughout the tree's lifespan.

Sexual and genotypic variation in terpene quantitative and qualitative profiles in the dioecious shrub Baccharis salicifolia.

Terpenoids are secondary metabolites produced in most plant tissues and are often considered toxic or repellent to plant enemies. Previous work has typically reported on intra-specific variation in terpene profiles, but the effects of plant sex, an important axis of genetic variation, have been less studied for chemical defences in general, and terpenes in particular. In a prior study, we found strong genetic variation (but not sexual dimorphism) in terpene amounts in leaves of the dioecious shrub Baccharis salicifolia. Here we build on these findings and provide a more in-depth analysis of terpene chemistry on these same plants from an experiment consisting of a common garden with male (N = 19) and female (N = 20) genotypes sourced from a single population. Our goal in the present study was to investigate quantitative and qualitative differences in terpene profiles associated with plant sex and genotypic variation. For this, we quantified leaf mono- and sesquiterpene amount, richness, and diversity (quantitative profile), as well as the composition of compounds (qualitative profile). We found no evidence of sexual dimorphism in monoterpene or sesquiterpene profiles. We did, however, find significant genotypic variation in amount, diversity, and composition of monoterpenes, but no effects on sesquiterpenes. These findings indicated that genotypic variation in terpene profiles largely surpassed variation due to sexual dimorphism for the studied population of this species.

Greater phylogenetic distance from native oaks predicts escape from insect leaf herbivores by non-native oak saplings.

PREMISE: Non-native plant species have been hypothesized to experience lower herbivory in novel environments as a function of their phylogenetic distance from native plant species. Although recent work has found support for this prediction, the plant traits responsible for such patterns have been largely overlooked. METHODS: In a common garden experiment in northwestern Spain, we tested whether oak species (Quercus spp.) not native to this region that are phylogenetically more distantly related to native species exhibit less insect leaf herbivory. In addition, we also investigated plant traits potentially correlated with any such effect of phylogenetic distance. RESULTS: As expected, phylogenetic distance from native species negatively predicted insect leaf herbivory on non-native oaks. In addition, we found that the leaf traits, namely phosphorus and condensed tannins, were significantly associated with herbivory, suggesting that they are associated with the effect of phylogenetic distance on leaf herbivory on non-native oak species. CONCLUSIONS: This study contributes to a better understanding of how evolutionary relationships (relatedness) between native and non-native plant species determine the latter's success in novel environments via locally shared enemies, and encourages more work investigating the plant traits that mediate the effects of phylogenetic distance on enemy escape.

Bottom-up control of geographic variation in insect herbivory on wild cotton (Gossypium hirsutum) by plant defenses and climate.

PREMISE: The occurrence and amount of herbivory are shaped by bottom-up forces, primarily plant traits (e.g., defenses), and by abiotic factors. Addressing these concurrent effects in a spatial context has been useful in efforts to understand the mechanisms governing variation in plant-herbivore interactions. Still, few studies have evaluated the simultaneous influence of multiple sources of bottom-up variation on spatial variation in herbivory. METHODS: We tested to what extent chemical (phenolics, production of terpenoid glands) and physical (pubescence) defensive plant traits and climatic factors are associated with variation in herbivory by leaf-chewing insects across populations of wild cotton (Gossypium hirsutum). RESULTS: We found substantial population variation in cotton leaf defenses and insect leaf herbivory. Leaf pubescence, but not gossypol gland density or phenolic content, was significantly negatively associated with herbivory by leaf-chewing insects. In addition, there were direct effects of climate on defenses and herbivory, with leaf pubescence increasing toward drier conditions and leaf damage increasing toward wetter and cooler conditions. There was no evidence, however, of indirect effects (via plant defenses) of climate on herbivory. CONCLUSIONS: These results suggest that spatial variation in insect herbivory on wild G. hirsutum is predominantly driven by concurrent and independent influences of population variation in leaf pubescence and climatic factors.

Neutral and Climate-Driven Adaptive Processes Contribute to Explain Population Variation in Resin Duct Traits in a Mediterranean Pine Species.

Resin ducts are important anatomical defensive traits related to biotic resistance in conifers. Previous studies have reported intraspecific genetic variation in resin duct characteristics. However, little is currently known about the micro-evolutionary patterns and adaptive value of these defensive structures. Here, we quantified inter-population genetic variation in resin duct features and their inducibility in Pinus pinaster and assessed whether such variation was associated with climate gradients. To that end, we characterized the resin duct system of 2-year-old saplings from 10 populations across the species' distribution range. We measured axial resin duct features (density, mean size, and percentage conductive area of resin ducts) and their inducibility in response to methyl jasmonate. Genotyping of single nucleotide polymorphisms allowed to account for the population genetic structure in our models in order to avoid spurious correlations between resin duct characteristics and climate. We found large inter-population variation in resin duct density and conductive area, but not in their inducibility. Our results suggest that population variation in the percentage conductive area of resin ducts likely arise from adaptation to local climate conditions. This study highlights the adaptive relevance of resin ducts and helps to shed light on the micro-evolutionary patterns of resin-based defenses in conifers.