The effect of local habitat and spatial connectivity on urban seed predation.

PREMISE: During the last centuries, the area covered by urban landscapes is increasing all over the world. Urbanization can change local habitats and decrease connectivity among these habitats, with important consequences for species interactions. While several studies have found a major imprint of urbanization on plant-insect interactions, the effects of urbanization on seed predation remain largely unexplored. METHODS: We investigated the relative impact of sunlight exposure, leaf litter, and spatial connectivity on predation by moth and weevil larvae on acorns of the pedunculate oak across an urban landscape during 2018 and 2020. We also examined whether infestations by moths and weevils were independent of each other. RESULTS: While seed predation varied strongly among trees, seed predation was not related to differences in sunlight exposure, leaf litter, or spatial connectivity. Seed predation by moths and weevils was negatively correlated at the level of individual acorns in 2018, but positively correlated at the acorn and the tree level in 2020. CONCLUSIONS: Our study sets the baseline expectation that urban seed predators are unaffected by differences in sunlight exposure, leaf litter, and spatial connectivity. Overall, our findings suggest that the impact of local and spatial factors on insects within an urban context may depend on the species guild. Understanding the impact of local and spatial factors on biodiversity, food web structure, and ecosystem functioning can provide valuable insights for urban planning and management strategies aimed at promoting urban insect diversity.

Parasitism rate differs between herbivore generations in the univoltine, but not bivoltine, range.

With climate change, plant-feeding insects increase their number of annual generations (voltinism). However, to what degree the emergence of a new herbivore generation affects the parasitism rate has not been explored. We performed a field experiment to test whether the parasitism rate differs between the first and the second generations of a specialist leaf miner (Tischeria ekebladella), both in the naturally univoltine and bivoltine parts of the leaf miner's distribution. We found an interactive effect between herbivore generation and geographical range on the parasitism rate. The parasitism rate was higher in the first compared to the second host generation in the part of the range that is naturally univoltine, whereas it did not differ between generations in the bivoltine range. Our experiment highlights that shifts in herbivore voltinism might release top-down control, with potential consequences for natural and applied systems.

Spring phenology and pathogen infection affect multigenerational plant attackers throughout the growing season.

Climate change has been shown to advance spring phenology, increase the number of insect generations per year (multivoltinism) and increase pathogen infection levels. However, we lack insights into the effects of plant spring phenology and the biotic environment on the preference and performance of multivoltine herbivores and whether such effects extend into the later part of the growing season. To this aim, we used a multifactorial growth chamber experiment to examine the influence of spring phenology on plant pathogen infection, and how the independent and interactive effects of spring phenology and plant pathogen infection affect the preference and performance of multigenerational attackers (the leaf miner Tischeria ekebladella and the aphid Tuberculatus annulatus) on the pedunculate oak in the early, mid and late parts of the plant growing season. Pathogen infection was highest on late phenology plants, irrespective of whether inoculations were conducted in the early, mid or late season. The leaf miner consistently preferred to oviposit on middle and late phenology plants, as well as healthy plants, during all parts of the growing season, whereas we detected an interactive effect between spring phenology and pathogen infection on the performance of the leaf miner. Aphids preferred healthy, late phenology plants during the early season, healthy plants during the mid season, and middle phenology plants during the late season, whereas aphid performance was consistently higher on healthy plants during all parts of the growing season. Our findings highlight that the impact of spring phenology on pathogen infection and the preference and performance of insect herbivores is not restricted to the early season, but that its imprint is still present - and sometimes equally strong - during the peak and end of the growing season. Plant pathogens generally negatively affected herbivore preference and performance, and modulated the effects of spring phenology. We conclude that spring phenology and pathogen infection are two important factors shaping the preference and performance of multigenerational plant attackers, which is particularly relevant given the current advance in spring phenology, pathogen outbreaks and increase in voltinism with climate change.

The co-existence of multiple oak leaf flushes contributes to the large within-tree variation in chemistry, insect attack and pathogen infection.

Many plant species produce multiple leaf flushes during the growing season, which might have major consequences for within-plant variation in chemistry and species interactions. Yet, we lack a theoretical or empirical framework for how differences among leaf flushes might shape variation in damage by insects and diseases. We assessed the impact of leaf flush identity on leaf chemistry, insect attack and pathogen infection on the pedunculate oak Quercus robur by sampling leaves from each leaf flush in 20 populations across seven European countries during an entire growing season. The first leaf flush had higher levels of primary compounds, and lower levels of secondary compounds, than the second flush, whereas plant chemistry was highly variable in the third flush. Insect attack decreased from the first to the third flush, whereas infection by oak powdery mildew was lowest on leaves from the first flush. The relationship between plant chemistry, insect attack and pathogen infection varied strongly among leaf flushes and seasons. Our findings demonstrate the importance of considering differences among leaf flushes for our understanding of within-tree variation in chemistry, insect attack and disease levels, something particularly relevant given the expected increase in the number of leaf flushes with climate change.

Changes in the foliar fungal community between oak leaf flushes along a latitudinal gradient in Europe.

Aim: Leaves support a large diversity of fungi, which are known to cause plant diseases, induce plant defences or influence leaf senescence and decomposition. To advance our understanding of how foliar fungal communities are structured and assembled, we assessed to what extent leaf flush and latitude can explain the within- and among-tree variation in foliar fungal communities. Location: A latitudinal gradient spanning c. 20 degrees in latitude in Europe. Taxa: The foliar fungal community associated with a foundation tree species, the pedunculate oak Quercus robur. Methods: We examined the main and interactive effects of leaf flush and latitude on the foliar fungal community by sampling 20 populations of the pedunculate oak Quercus robur across the tree's range. We used the ITS region as a target for characterization of fungal communities using DNA metabarcoding. Results: Species composition, but not species richness, differed between leaf flushes. Across the latitudinal gradient, species richness was highest in the central part of the oak's distributional range, and foliar fungal community composition shifted along the latitudinal gradient. Among fungal guilds, the relative abundance of plant pathogens and mycoparasites was lower on the first leaf flush, and the relative abundance of plant pathogens and saprotrophs decreased with latitude. Conclusions: Changes in community composition between leaf flushes and along the latitudinal gradient were mostly a result of species turnover. Overall, our findings demonstrate that leaf flush and latitude explain 5%-22% of the small- and large-scale spatial variation in the foliar fungal community on a foundation tree within the temperate region. Using space-for-time substitution, we expect that foliar fungal community structure will change with climate warming, with an increase in the abundance of plant pathogens and mycoparasites at higher latitudes, with major consequences for plant health, species interactions and ecosystem dynamics.

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.

DNA Barcoding and geographical scale effect: The problems of undersampling genetic diversity hotspots.

DNA barcoding identification needs a good characterization of intraspecific genetic divergence to establish the limits between species. Yet, the number of barcodes per species is many times low and geographically restricted. A poor coverage of the species distribution range may hamper identification, especially when undersampled areas host genetically distinct lineages. If so, the genetic distance between some query sequences and reference barcodes may exceed the maximum intraspecific threshold for unequivocal species assignation. Taking a group of Quercus herbivores (moths) in Europe as model system, we found that the number of DNA barcodes from southern Europe is proportionally very low in the Barcoding of Life Data Systems. This geographical bias complicates the identification of southern query sequences, due to their high intraspecific genetic distance with respect to barcodes from higher latitudes. Pairwise intraspecific genetic divergence increased along with spatial distance, but was higher when at least one of the sampling sites was in southern Europe. Accordingly, GMYC (General Mixed Yule Coalescent) single-threshold model retrieved clusters constituted exclusively by Iberian haplotypes, some of which could correspond to cryptic species. The number of putative species retrieved was more reliable than that of multiple-threshold GMYC but very similar to results from ABGD and jMOTU. Our results support GMYC as a key resource for species delimitation within poorly inventoried biogeographic regions in Europe, where historical factors (e.g., glaciations) have promoted genetic diversity and singularity. Future European DNA barcoding initiatives should be preferentially performed along latitudinal gradients, with special focus on southern peninsulas.

Latitudinal variation in seed predation correlates with latitudinal variation in seed defensive and nutritional traits in a widespread oak species.

BACKGROUND AND AIMS: Classic theory on geographical gradients in plant-herbivore interactions assumes that herbivore pressure and plant defences increase towards warmer and more stable climates found at lower latitudes. However, the generality of these expectations has been recently called into question by conflicting empirical evidence. One possible explanation for this ambiguity is that most studies have reported on patterns of either herbivory or plant defences whereas few have measured both, thus preventing a full understanding of the implications of observed patterns for plant-herbivore interactions. In addition, studies have typically not measured climatic factors affecting plant-herbivore interactions, despite their expected influence on plant and herbivore traits. METHODS: Here we tested for latitudinal variation in insect seed predation and seed traits putatively associated with insect attack across 36 Quercus robur populations distributed along a 20° latitudinal gradient. We then further investigated the associations between climatic factors, seed traits and seed predation to test for climate-based mechanisms of latitudinal variation in seed predation. KEY RESULTS: We found strong but contrasting latitudinal clines in seed predation and seed traits, whereby seed predation increased whereas seed phenolics and phosphorus decreased towards lower latitudes. We also found a strong direct association between temperature and seed predation, with the latter increasing towards warmer climates. In addition, temperature was negatively associated with seed traits, with populations at warmer sites having lower levels of total phenolics and phosphorus. In turn, these negative associations between temperature and seed traits led to a positive indirect association between temperature and seed predation. CONCLUSIONS: These results help unravel how plant-herbivore interactions play out along latitudinal gradients and expose the role of climate in driving these outcomes through its dual effects on plant defences and herbivores. Accordingly, this emphasizes the need to account for abiotic variation while testing concurrently for latitudinal variation in plant traits and herbivore pressure.

Seed loss before seed predation: experimental evidence of the negative effects of leaf feeding insects on acorn production.

Insect herbivory decreases plant fitness by constraining plant growth, survival and reproductive output. Most studies on the effects of herbivory in trees rely on correlational inter-individual comparisons and could, thus, be affected by confounding factors linked to both herbivory and plant performance. Using the Mediterranean Holm oak (Quercus ilex) as a study model, we followed an experimental approach in which leaf-feeding insects (mainly Lepidoptera caterpillars) were excluded from some shoots in all study trees. Shoots subjected to herbivore exclusion exhibited lower defoliation rates and produced more acorns than control shoots. Defoliation constrained shoot growth throughout the study period, but had no effect on the number of female flowers produced per shoot. Acorn production was, however, lower in control shoots due to their higher abortion rates, and also to their greater mortality risk during summer drought, as shoots with fewer leaves were less likely to survive. Plant reaction to herbivory inhibits certain physiological pathways involved in plant growth, which, together with the effects of physical damage, reduces the amount and efficiency of the photosynthetic tissue. This increases their vulnerability to environmental stresses, such as water deficit, which limit resource assimilation. Defoliation is likely a key factor affecting oak regeneration, as it may be a significant source of seed loss prior to pre-dispersal acorn predation. Further experimental studies could help to elucidate its effects in contrasting environments. In Mediterranean regions, the harsher droughts predicted by climate change models could worsen the effects of insect herbivory on oak reproductive output.