Vertically stratified interactions of nectarivores and nectar-inhabiting bacteria in a liana flowering across forest strata.

PREMISE: Vertical stratification is a key feature of tropical forests and plant-frugivore interactions. However, it is unclear whether equally strong patterns of vertical stratification exist for plant-nectarivore interactions and, if so, which factors drive these patterns. Further, nectar-inhabiting bacteria, acting as "hidden players" in plant-nectarivore interactions, might be vertically stratified, either in response to differences among strata in microenvironmental conditions or to the nectarivore community serving as vectors. METHODS: We observed visitations by a diverse nectarivore community to the liana Marcgravia longifolia in a Peruvian rainforest and characterized diversity and community composition of nectar-inhabiting bacteria. Unlike most other plants, M. longifolia produces inflorescences across forest strata, enabling us to study effects of vertical stratification on plant-nectarivore interactions without confounding effects of plant species and stratum. RESULTS: A significantly higher number of visits were by nectarivorous bats and hummingbirds in the midstory than in the understory and canopy, and the visits were strongly correlated to flower availability and nectar quantity and quality. Trochiline hummingbirds foraged across all strata, whereas hermits remained in the lower strata. The Shannon diversity index for nectar-inhabiting bacterial communities was highest in the midstory. CONCLUSIONS: Our findings suggest that vertical niche differentiation in plant-nectarivore interactions seems to be partly driven by resource abundance, but other factors such as species-specific preferences of hummingbirds, likely caused by competition, play an important role. We conclude that vertical stratification is an important driver of a species' interaction niche highlighting its role for promoting biodiversity and ecosystem functioning.

Adding experimental precision to the realism of field observations: Plant communities structure bacterial communities in a glacier forefield.

Ecological studies are aligned along a realism-precision continuum ranging from field observations to controlled lab experiments that each have their own strengths and limitations. Ecological insight may be most robust when combining approaches. In field observations along a successional gradient, we found correlations between plant species composition and soil bacterial communities, while bacterial Shannon diversity was unrelated to vegetation characteristics. To add a causal understanding of the processes of bacterial community assembly, we designed lab experiments to specifically test the influence of plant composition on bacterial communities. Using soil and seeds from our field site, we added different combinations of surface-sterilised seeds to homogenised soil samples in microcosms and analysed bacterial communities 4 months later. Our results confirmed the field observations suggesting that experimental plant community composition shaped bacterial community composition, while Shannon diversity was unaffected. These results reflect intimate plant-bacteria interactions that are important drivers of plant health and community assembly. While this study provided insights into the role of plants underlying the assembly of bacterial communities, we did not experimentally manipulate other drivers of community assembly such as abiotic factors. Therefore, we recommend multi-factorial laboratory experiments to quantify the relative importance of different factors contributing to microbial composition.

Exploring the Frequency and Distribution of Ecological Non-monotonicity in Associations among Ecosystem Constituents.

Complex links between biotic and abiotic constituents are fundamental for the functioning of ecosystems. Although non-monotonic interactions and associations are known to increase the stability, diversity, and productivity of ecosystems, they are frequently ignored by community-level standard statistical approaches. Using the copula-based dependence measure qad, capable of quantifying the directed and asymmetric dependence between variables for all forms of (functional) relationships, we determined the proportion of non-monotonic associations between different constituents of an ecosystem (plants, bacteria, fungi, and environmental parameters). Here, we show that up to 59% of all statistically significant associations are non-monotonic. Further, we show that pairwise associations between plants, bacteria, fungi, and environmental parameters are specifically characterized by their strength and degree of monotonicity, for example, microbe-microbe associations are on average stronger than and differ in degree of non-monotonicity from plant-microbe associations. Considering directed and non-monotonic associations, we extended the concept of ecosystem coupling providing more complete insights into the internal order of ecosystems. Our results emphasize the importance of ecological non-monotonicity in characterizing and understanding ecosystem patterns and processes. Supplementary Information: The online version contains supplementary material available at 10.1007/s10021-023-00867-9.

The great escape: patterns of enemy release are not explained by time, space or climate.

When a plant is introduced to a new ecosystem it may escape from some of its coevolved herbivores. Reduced herbivore damage, and the ability of introduced plants to allocate resources from defence to growth and reproduction can increase the success of introduced species. This mechanism is known as enemy release and is known to occur in some species and situations, but not in others. Understanding the conditions under which enemy release is most likely to occur is important, as this will help us to identify which species and habitats may be most at risk of invasion. We compared in situ measurements of herbivory on 16 plant species at 12 locations within their native European and introduced Australian ranges to quantify their level of enemy release and understand the relationship between enemy release and time, space and climate. Overall, plants experienced approximately seven times more herbivore damage in their native range than in their introduced range. We found no evidence that enemy release was related to time since introduction, introduced range size, temperature, precipitation, humidity or elevation. From here, we can explore whether traits, such as leaf defences or phylogenetic relatedness to neighbouring plants, are stronger indicators of enemy release across species.

Fungicides and strawberry pollination-Effects on floral scent, pollen attributes and bumblebee behavior.

Fungicides are used in agriculture to protect crops from various fungal diseases. However, they may modulate the plants metabolism. Moreover, fungicides can accumulate in the environment and may cause toxic effects on non-target organisms such as nectar microbes and pollinators. Nectar microbes contribute to the volatile profile of flowers and can influence pollinators behaviour. Thus, fungicide treatment could potentially affect the pollination. In this study, we investigated the influence of fungicide treatment on floral attributes as well as the behavioural impact on bumblebees. In separate experiments, we used one or both strawberry cultivars (Fragaria × ananassa var. Darselect and Malwina), which were either kept untreated (control) or treated with either Cuprozin® progress or SWITCH® fungicide. We analysed various flower traits including volatiles, pollen weight, pollen protein, and the attraction of bumblebees towards the flowers in the greenhouse. Additionally, we analysed the viability of pollen and pollen live-to-dead ratio, as well as the composition of nectar fungi in the field. A treatment with Cuprozin® progress led to a lower emission of floral volatiles and a slightly lower pollen protein content. This had no impact on the visit latency of bumblebees but on the overall visit frequency of these flowers. The treatment with the fungicide SWITCH® resulted in a higher emission of floral volatiles as well as a delayed first visit by bumblebees. Furthermore, flowers of control plants were visited more often than those treated with the two fungicides. Plant-pollinator interactions are highly complex, with many contributing factors. Fungicides can have an impact on the pollen quality and pollinator attraction, potentially leading to an altered pollen dispersal by pollinators and a change in fruit quality.

Digital whole-community phenotyping: tracking morphological and physiological responses of plant communities to environmental changes in the field.

Plant traits are informative for ecosystem functions and processes and help to derive general rules and predictions about responses to environmental gradients, global change and perturbations. Ecological field studies often use 'low-throughput' methods to assess plant phenotypes and integrate species-specific traits to community-wide indices. In contrast, agricultural greenhouse or lab-based studies often employ 'high-throughput phenotyping' to assess plant individuals tracking their growth or fertilizer and water demand. In ecological field studies, remote sensing makes use of freely movable devices like satellites or unmanned aerial vehicles (UAVs) which provide large-scale spatial and temporal data. Adopting such methods for community ecology on a smaller scale may provide novel insights on the phenotypic properties of plant communities and fill the gap between traditional field measurements and airborne remote sensing. However, the trade-off between spatial resolution, temporal resolution and scope of the respective study requires highly specific setups so that the measurements fit the scientific question. We introduce small-scale, high-resolution digital automated phenotyping as a novel source of quantitative trait data in ecological field studies that provides complementary multi-faceted data of plant communities. We customized an automated plant phenotyping system for its mobile application in the field for 'digital whole-community phenotyping' (DWCP), capturing the 3-dimensional structure and multispectral information of plant communities. We demonstrated the potential of DWCP by recording plant community responses to experimental land-use treatments over two years. DWCP captured changes in morphological and physiological community properties in response to mowing and fertilizer treatments and thus reliably informed about changes in land-use. In contrast, manually measured community-weighted mean traits and species composition remained largely unaffected and were not informative about these treatments. DWCP proved to be an efficient method for characterizing plant communities, complements other methods in trait-based ecology, provides indicators of ecosystem states, and may help to forecast tipping points in plant communities often associated with irreversible changes in ecosystems.

Floral volatiles evoke partially similar responses in both florivores and pollinators and are correlated with non-volatile reward chemicals.

BACKGROUND: Plants often use floral displays to attract mutualists and prevent antagonist attacks. Chemical displays detectable from a distance include attractive or repellent floral volatile organic compounds (FVOCs). Locally, visitors perceive contact chemicals including nutrients but also deterrent or toxic constituents of pollen and nectar. The FVOC and pollen chemical composition can vary intra- and interspecifically. For certain pollinator and florivore species, responses to these compounds are studied in specific plant systems, yet we lack a synthesis of general patterns comparing these two groups and insights into potential correlations between FVOC and pollen chemodiversity. SCOPE: We reviewed how FVOCs and non-volatile floral chemical displays, i.e. pollen nutrients and toxins, vary in composition and affect the detection by and behaviour of insect visitors. Moreover, we used meta-analyses to evaluate the detection of and responses to FVOCs by pollinators vs. florivores within the same plant genera. We also tested whether the chemodiversity of FVOCs, pollen nutrients and toxins is correlated, hence mutually informative. KEY RESULTS: According to available data, florivores could detect more FVOCs than pollinators. Frequently tested FVOCs were often reported as pollinator-attractive and florivore-repellent. Among FVOCs tested on both visitor groups, there was a higher number of attractive than repellent compounds. FVOC and pollen toxin richness were negatively correlated, indicating trade-offs, whereas a marginal positive correlation between the amount of pollen protein and toxin richness was observed. CONCLUSIONS: Plants face critical trade-offs, because floral chemicals mediate similar information to both mutualists and antagonists, particularly through attractive FVOCs, with fewer repellent FVOCs. Furthermore, florivores might detect more FVOCs, whose richness is correlated with the chemical richness of rewards. Chemodiversity of FVOCs is potentially informative of reward traits. To gain a better understanding of the ecological processes shaping floral chemical displays, more research is needed on floral antagonists of diverse plant species and on the role of floral chemodiversity in visitor responses.

Inter- and intraspecific phytochemical variation correlate with epiphytic flower and leaf bacterial communities.

Microbes associated with flowers and leaves affect plant health and fitness and modify the chemical phenotypes of plants with consequences for interactions of plants with their environment. However, the drivers of bacterial communities colonizing above-ground parts of grassland plants in the field remain largely unknown. We therefore examined the relationships between phytochemistry and the epiphytic bacterial community composition of flowers and leaves of Ranunculus acris and Trifolium pratense. On 252 plant individuals, we characterized primary and specialized metabolites, that is, surface sugars, volatile organic compounds (VOCs), and metabolic fingerprints, as well as epiphytic flower and leaf bacterial communities. The genomic potential of bacterial colonizers concerning metabolic capacities was assessed using bacterial reference genomes. Phytochemical composition displayed pronounced variation within and between plant species and organs, which explained part of the variation in bacterial community composition. Correlation network analysis suggests strain-specific correlations with metabolites. Analysis of bacterial reference genomes revealed taxon-specific metabolic capabilities that corresponded with genes involved in glycolysis and adaptation to osmotic stress. Our results show relationships between phytochemistry and the flower and leaf bacterial microbiomes suggesting that plants provide chemical niches for distinct bacterial communities. In turn, bacteria may induce alterations in the plants' chemical phenotype. Thus, our study may stimulate further research on the mechanisms of trait-based community assembly in epiphytic bacteria.

Quantifying chemodiversity considering biochemical and structural properties of compounds with the R package chemodiv.

Plants produce large numbers of phytochemical compounds affecting plant physiology and interactions with their biotic and abiotic environment. Recently, chemodiversity has attracted considerable attention as an ecologically and evolutionary meaningful way to characterize the phenotype of a mixture of phytochemical compounds. Currently used measures of phytochemical diversity, and related measures of phytochemical dissimilarity, generally do not take structural or biosynthetic properties of compounds into account. Such properties can be indicative of the compounds' function and inform about their biosynthetic (in)dependence, and should therefore be included in calculations of these measures. We introduce the R package chemodiv, which retrieves biochemical and structural properties of compounds from databases and provides functions for calculating and visualizing chemical diversity and dissimilarity for phytochemicals and other types of compounds. Our package enables calculations of diversity that takes the richness, relative abundance and - most importantly - structural and/or biosynthetic dissimilarity of compounds into account. We illustrate the use of the package with examples on simulated and real datasets. By providing the R package chemodiv for quantifying multiple aspects of chemodiversity, we hope to facilitate investigations of how chemodiversity varies across levels of biological organization, and its importance for the ecology and evolution of plants and other organisms.

Sugar Concentration, Nitrogen Availability, and Phylogenetic Factors Determine the Ability of Acinetobacter spp. and Rosenbergiella spp. to Grow in Floral Nectar.

The floral nectar of angiosperms harbors a variety of microorganisms that depend predominantly on animal visitors for their dispersal. Although some members of the genus Acinetobacter and all currently known species of Rosenbergiella are thought to be adapted to thrive in nectar, there is limited information about the response of these bacteria to variation in the chemical characteristics of floral nectar. We investigated the growth performance of a diverse collection of Acinetobacter (n = 43) and Rosenbergiella (n = 45) isolates obtained from floral nectar and the digestive tract of flower-visiting bees in a set of 12 artificial nectars differing in sugar content (15% w/v or 50% w/v), nitrogen content (3.48/1.67 ppm or 348/167 ppm of total nitrogen/amino nitrogen), and sugar composition (only sucrose, 1/3 sucrose + 1/3 glucose + 1/3 fructose, or 1/2 glucose + 1/2 fructose). Growth was only observed in four of the 12 artificial nectars. Those containing elevated sugar concentration (50% w/v) and low nitrogen content (3.48/1.67 ppm) were limiting for bacterial growth. Furthermore, phylogenetic analyses revealed that the ability of the bacteria to grow in different types of nectar is highly conserved between closely related isolates and genotypes, but this conservatism rapidly vanishes deeper in phylogeny. Overall, these results demonstrate that the ability of Acinetobacter spp. and Rosenbergiella spp. to grow in floral nectar largely depends on nectar chemistry and bacterial phylogeny.

Floral and reproductive traits are an independent dimension within the plant economic spectrum of temperate central Europe.

Major dimensions of plant ecological strategies have been widely studied bringing forward the concept of 'economic spectra' of plants. Sexual reproductive traits, 'floral traits', have been largely neglected in this context, despite their strong link to fitness. Here, we aimed at integrating floral traits into the dimensionality of plant form and function so far dominated by vegetative traits. We used principal component analyses and constructed trait networks to assess the correlation structure of leaf, belowground, plant size-related, and floral traits. We studied forbs within two independent datasets; one compiled from central European trait databases and one sampled in the Austrian Alps. Floral traits defined the second dimension of trait variability within both datasets, while plant size determined the first dimension. Floral traits were largely independent from the leaf economic spectrum. Flower size, however, positively scaled with plant size and leaf size. Mating system was the most well-connected trait across modules of plant tissue/organ types. The independence of floral traits was consistent also after accounting for phylogenetic relationships between species. Floral traits explained a unique part of the variation in plant form and function and thus, likely play a distinctive ecological role within the whole plant economic spectrum.

Succession comprises a sequence of threshold-induced community assembly processes towards multidiversity.

Research on successions and community assembly both address the same processes such as dispersal, species sorting, and biotic interactions but lack unifying concepts. Recent theoretical advances integrated both research lines proposing a sequence of stochastic and deterministic processes along successional gradients. Shifts in ecosystem states along successional gradients are predicted to occur abruptly once abiotic and biotic factors dominate over dispersal as main driver. Considering the multidiversity composed of five organismal groups including plants, animals, and microbes, our results imply that stochastic, likely dispersal-dominated, processes are replaced by rather deterministic processes such as environmental filtering and biotic interactions after around 60 years of succession in a glacier forefield. The niche-based character of later successional processes is further supported by a decline in multi-beta-diversity. Our results may update concepts of community assembly by considering multiple taxa, help to bridge the gap between research on successions and community assembly, and provide insights into the emergence of multidiverse and complex ecosystems.

Chemical phenotype as important and dynamic niche dimension of plants.

Niche theory considering the traits of species and individuals provides a powerful tool to integrate ecology and evolution of species. In plant ecology, morphological and physiological traits are commonly considered as niche dimensions, whereas phytochemical traits are mostly neglected in this context despite their pivotal functions in plant responses to their environment and in mediating interactions. The diversity of plant phytochemicals can thus mediate three key processes: niche choice, conformance and construction. Here, we integrate frameworks from niche theory with chemical ecology and argue that plants use their individual-specific diversity in phytochemicals (chemodiversity) for different niche realization processes. Our concept has important implications for ecosystem processes and stability and increases the predictive ability of chemical ecology.

Accuracy of mutual predictions of plant and microbial communities vary along a successional gradient in an alpine glacier forefield.

Receding glaciers create virtually uninhabited substrates waiting for initial colonization of bacteria, fungi and plants. These glacier forefields serve as an ideal ecosystem for studying transformations in community composition and diversity over time and the interactions between taxonomic groups in a dynamic landscape. In this study, we investigated the relationships between the composition and diversity of bacteria, fungi, and plant communities as well as environmental factors along a successional gradient. We used random forest analysis assessing how well the composition and diversity of taxonomic groups and environmental factors mutually predict each other. We did not identify a single best indicator for all taxonomic and environmental properties, but found specific predictors to be most accurate for each taxon and environmental factor. The accuracy of prediction varied considerably along the successional gradient, highlighting the dynamic environmental conditions along the successional gradient that may also affect biotic interactions across taxa. This was also reflected by the high accuracy of predictions of plot age by all taxa. Next to plot age, our results indicate a strong importance of pH and temperature in structuring microbial and plant community composition. In addition, taxonomic groups predicted the community composition of each other more accurately than environmental factors, which may either suggest that these groups similarly respond to other not measured environmental factors or that direct interactions between taxa shape the composition of their communities. In contrast, diversity of taxa was not well predicted, suggesting that community composition of one taxonomic group is not a strong driver of the diversity of another group. Our study provides insights into the successional development of multidiverse communities shaped by complex interactions between taxonomic groups and the environment.

Effect of Tannin Furanic Polymer in Comparison to Its Mimosa Tannin Extract on the Growth of Bacteria and White-Rot Fungi.

Tannins are well-known to protect plants from bacteria and fungi, but nothing is known about its effects on microorganisms once they are copolymerized. Therefore, a study was conducted to evaluate the effect of a tannin−furanic polymer in comparison with industrial mimosa tannin extract on the in vitro growth of two strains of bacteria, Bacillaceae and Pseudomanadaceae, and two white-rot fungi, Trametes versicolor and Agrocybe aegerita. Results have highlighted that the tannin polymer did not inhibit the growth of tested bacteria and even favored the growth of Bacillaceae without extra glucose. The growth of both fungi was enhanced by mimosa tannin and its polymer at low concentrations (<1%), while concentrations above 10% had a growth-inhibiting effect, which was slightly less strong for the polymer compared to the tannin against Trametes versicolor. These findings highlighted that tannin−furanic polymers can be tolerated by certain microorganisms at low concentration and that their inhibitory effect is similar or slightly lower than that of the pristine tannin extract.

Divergent assembly processes? A comparison of the plant and soil microbiome with plant communities in a glacier forefield.

Community assembly is a result of dispersal, abiotic and biotic characteristics of the habitat as well as stochasticity. A direct comparison between the assembly of microbial and 'macrobial' organisms is hampered by the sampling of these communities in different studies, at different sites or on different scales. In a glacier forefield in the Austrian Alps, we recorded the soil and plant microbiome (bacteria and fungi) and plants that occurred in the same landscape and in close proximity in the same plots. We tested five predictions deduced from assembly processes and revealed deviating patterns of assembly in these community types. In short, microbes appeared to be less dispersal limited than plants and soil microbes, and plants strongly responded to abiotic factors whereas the leaf microbiome was plant species specific and well buffered from environmental conditions. The observed differences in community assembly processes may be attributed to the organisms' dispersal abilities, the exposure of the habitats to airborne propagules and habitat characteristics. The finding that assembly is conditional to the characteristics of the organisms, the habitat and the spatial scale under consideration is thus central for our understanding about the establishment and the maintenance of biodiversity.

Humans Share More Preferences for Floral Phenotypes With Pollinators Than With Pests.

Studies on the selection of floral traits usually consider pollinators and sometimes herbivores. However, humans also exert selection on floral traits of ornamental plants. We compared the preferences of bumblebees (Bombus terrestris), thrips (Frankliniella occidentalis), and humans for flowers of snapdragon. From a cross of two species, Antirrhinum majus and Antirrhinum linkianum, we selected four Recombinant Inbred Lines (RILs). We characterised scent emission from whole flowers and stamens, pollen content and viability, trichome density, floral shape, size and colour of floral parts. We tested the preferences of bumblebees, thrips, and humans for whole flowers, floral scent bouquets, stamen scent, and individual scent compounds. Humans and bumblebees showed preferences for parental species, whereas thrips preferred RILs. Colour and floral scent, in combination with other floral traits, seem relevant phenotypes for all organisms. Remarkably, visual traits override scent cues for bumblebees, although, scent is an important trait when bumblebees cannot see the flowers, and methyl benzoate was identified as a key attractant for them. The evolutionary trajectory of flowers is the result of multiple floral traits interacting with different organisms with different habits and modes of interaction.

Volatile organic compound patterns predict fungal trophic mode and lifestyle.

Fungi produce a wide variety of volatile organic compounds (VOCs), which play central roles in the initiation and regulation of fungal interactions. Here we introduce a global overview of fungal VOC patterns and chemical diversity across phylogenetic clades and trophic modes. The analysis is based on measurements of comprehensive VOC profiles of forty-three fungal species. Our data show that the VOC patterns can describe the phyla and the trophic mode of fungi. We show different levels of phenotypic integration (PI) for different chemical classes of VOCs within distinct functional guilds. Further computational analyses reveal that distinct VOC patterns can predict trophic modes, (non)symbiotic lifestyle, substrate-use and host-type of fungi. Thus, depending on trophic mode, either individual VOCs or more complex VOC patterns (i.e., chemical communication displays) may be ecologically important. Present results stress the ecological importance of VOCs and serve as prerequisite for more comprehensive VOCs-involving ecological studies.