An experimental, behavioral, and chemical analysis of food limitations in mutualistic Crematogaster ant symbionts inhabiting Macaranga host plants.

Obligate mutualistic plant-ants are often constrained by their plant partner's capacity to provide resources. However, despite this limitation, some ant partners actively reject potential prey items and instead drop them from the plant rather than consuming them, leaving the ants entirely reliant on host plant-provided food, including that provided indirectly by the symbiotic scale insects that ants tend inside the plants. This dependency potentially increases the efficiency of these ants in defending their host. We hypothesize that if this ant behavior was beneficial to the symbiosis, prey rejection by ants would be observed across multiple plant host species. We also hypothesize that plant-provided food items and symbiotic scale insects from other ant plants should be rejected. We address these hypotheses in the Crematogaster ant-Macaranga plant system, in which plants provide living space and food, while ants protect plants from herbivory. We observed food acceptance and rejection behavior across five ant species and three plant host species. Ants were offered three types of food: termites as a surrogate herbivore, symbiotic scale insects, and nutritious food bodies (FB) produced by different host plant species. The unique ant species living in M. winkleri was the most likely to reject food items not provided by the plant species, followed by ants in M. glandibracteolata, while ants in M. pearsonii accepted most items offered to them. Using stable isotopes, chemical cues, and proteomic analyses, we demonstrate that this behavior was not related to differences between plant species in nutritional quality or composition of FB. Isotopic signatures revealed that certain species are primary consumers but other ant species can be secondary consumers even where surrogate herbivores are rejected, although these values varied depending on the ant developmental stage and plant species. Macaranga pearsonii and M. glandibracteolata, the two most closely related plant species, had most similar surface chemical cues of FB. However, M. glandibracteolata had strongest differences in food body nutritional content, isotopic signatures, and protein composition from either of the other two plant species studied. Taken together we believe our results point toward potential host coercion of symbiont ants by plants in the genus Macaranga Thouars (Euphorbiaceae).

Short-term plasticity and variation in acacia ant-rewards under different conditions of ant occupancy and herbivory.

In ant-plant defense mutualisms, plants known as myrmecophytes provide food and shelter to ant partners in exchange for defense against herbivores and pathogens. To ensure interaction pay-off, myrmecophytes must regulate their investment in ant-rewards depending on local conditions and herbivore pressure. We investigated how myrmecophyte investment in multiple ant-rewards relates to herbivory, ant defense, and ant occupancy over time. Specifically, we examined the plasticity of ant-rewards produced by swollen-thorn acacias (Vachellia collinsii) under different ant occupancy and herbivory conditions. We compared food rewards (number of extrafloral nectaries and pinnules as a proxy for food bodies) and housing rewards (domatia dimensions) of V. collinsii for three conditions: (1) occupied (defended by the obligate mutualist Pseudomyrmex spinicola) versus unoccupied trees, (2) occupied trees subject to an experimental herbivory manipulation versus control trees, and (3) trees occupied by different ant species varying in their level of defense (P. spinicola, P. simulans, Crematogaster crinosa). We found that food rewards were more likely to vary in time depending on ant occupancy and resident species. Conversely, housing rewards varied with the condition (occupancy or species of partner) and less through time. A one-time herbivory manipulation did not cause any changes to the ant-rewards produced. Our results reveal short-term plasticity in V. collinsii ant-rewards and demonstrate that myrmecophytes with constitutive rewards can adjust their investment in ant-rewards depending on the presence and identity of ant partners.

Morphological characterization of domatium development in Callicarpa saccata.

BACKGROUND AND AIMS: Domatia are plant structures within which organisms reside. Callicarpa saccata (Lamiaceae) is the sole myrmecophyte, or 'ant plant', that develops foliar (leaf-borne) myrmeco-domatia in this genus. In this work we examined domatium development in C. saccata to understand the developmental processes behind pouch-like domatia. METHODS: Scanning electron microscopy, sectioning and microcomputed tomography were carried out to compare the leaves of C. saccata with those of the closely related but domatia-less myrmecophyte Callicarpa subaequalis, both under cultivation without ants. KEY RESULTS: Callicarpa saccata domatia are formed as a result of excess cell proliferation at the blade/petiole junctions of leaf primordia. Blade/petiole junctions are important meristematic sites in simple leaf organogenesis. We also found that the mesophyll tissue of domatia does not clearly differentiate into palisade and spongy layers. CONCLUSIONS: Rather than curling of the leaf margins, a perturbation of the normal functioning of the blade/petiole junction results in the formation of domatium tissue. Excess cell proliferation warps the shape of the blade and disturbs the development of the proximal-distal axis. This process leads to the generation of distinct structures that facilitate interaction between C. saccata and ants.

Azteca ants maintain unique microbiomes across functionally distinct nest chambers.

The microbiome of built structures has considerable influence over an inhabitant's well-being, yet the vast majority of research has focused on human-built structures. Ants are well-known architects, capable of constructing elaborate dwellings, the microbiome of which is underexplored. Here, we explore the bacterial and fungal microbiomes in functionally distinct chambers within and outside the nests of Azteca alfari ants in Cecropia peltata trees. We predicted that A. alfari colonies (1) maintain distinct microbiomes within their nests compared to the surrounding environment, (2) maintain distinct microbiomes among nest chambers used for different functions, and (3) limit both ant and plant pathogens inside their nests. In support of these predictions, we found that internal and external nest sampling locations had distinct microbial communities, and A. alfari maintained lower bacterial richness in their 'nurseries'. While putative animal pathogens were suppressed in chambers that ants actively inhabited, putative plant pathogens were not, which does not support our hypothesis that A. alfari defends its host trees against microbial antagonists. Our results show that ants influence microbial communities inside their nests similar to studies of human homes. Unlike humans, ants limit the bacteria in their nurseries and potentially prevent the build-up of insect-infecting pathogens. These results highlight the importance of documenting how indoor microbiomes differ among species, which might improve our understanding of how to promote indoor health in human dwellings.

An Epiphytic Ant-Plant Mutualism Structures Arboreal Ant Communities.

Arboreal ant communities are primarily structured by interactions among ant species, food availability, and physical structures within the environment. Epiphytes are a common feature of tropical forests that can provide ants with both food and nesting space. To date, little work has examined what role epiphytic ant-plants play in structuring arboreal ant communities. We surveyed ant species inhabiting the Australian epiphytic ant-plant Myrmecodia beccarii Hook.f. (Gentianales: Rubiaceae) and how arboreal ant communities are structured in relation to M. beccarii presence on trees. Myrmecodia beccarii was inhabited by the ant Philidris cordata Smith, F. (Hymenoptera: Formicidae) on the majority of Melaleuca viridiflora Sol. Ex Gaertn. (Myrtales: Myrtaceae) trees with ant-occupied ant-plants at our two sites. Dominant arboreal ant species at both study sites exhibited discrete, nonoverlapping distributions, and C-score analysis detected an ant mosaic at one site. The distribution of P. cordata was limited by the distribution of ant-plants for both sites. Philidris cordata dominance on trees was also determined by the presence of M. beccarii occupied by P. cordata at both sites. We suggest that by providing P. cordata with nesting space M. beccarii plays a role in structuring these arboreal ant communities.

Taxonomy of coccids (Hemiptera: Coccidae: Coccus L.) associated with Crematogaster ants (Hymenoptera: Formicidae) in the stems of Macaranga plants (Euphorbiaceae) in Southeast Asia.

The Southeast Asian soft scale insects (Hemiptera: Coccomorpha: Coccidae) associated with ants of the Crematogaster borneensis-group (Hymenoptera: Formicidae) and living in the hollow stems of Macaranga plants (Euphorbiaceae) are revised taxonomically. Ten species of the genus Coccus L. are recognised: seven were described previously and three new species are described herein. The species are: Coccus caviramicolus Morrison, C. circularis Morrison, C. heckrothi Gullan Kondo sp. n., C. lambirensis Gullan Kondo sp. n., C. macarangicolus Takahashi, C. macarangae Morrison, C. penangensis Morrison, C. pseudotumuliferus Gullan Kondo sp. n., C. secretus Morrison and C. tumuliferus Morrison. All of these species are described or redescribed and newly illustrated based on the adult females, and a key to distinguish the species is provided. We designate a lectotype for C. macarangicolus. The first-instar nymphs of all species are morphologically extremely similar and therefore only the first-instar nymph of C. macarangae is described and illustrated. Seven of these species currently are known only from Macaranga, but C. macarangae, C. secretus and perhaps C. pseudotumuliferus have been recorded from the hollow stems of several other ant-plants and a few non-myrmecophytes. The Coccus species from Macaranga are closely related to C. hesperidum L., the type species of the genus, and therefore are retained in the genus Coccus even though the adult females exhibit a few morphological differences from C. hesperidum. The species of Coccus from Macaranga appear to be parthenogenetic because no male nymphs or adults have been found, despite extensive collecting.

Economy of scale: third partner strengthens a keystone ant-plant mutualism.

While foundation species can stabilize ecosystems at landscape scales, their ability to persist is often underlain by keystone interactions occurring at smaller scales. Acacia drepanolobium is a foundation tree, comprising >95% of woody cover in East African black-cotton savanna ecosystems. Its dominance is underlain by a keystone mutualistic interaction with several symbiotic ant species in which it provides housing (swollen thorns) and carbohydrate-rich nectar from extra-floral nectaries (EFN). In return, it gains protection from catastrophic damage from mega-herbivores. Crematogaster mimosae is the ecologically dominant symbiotic ant in this system, also providing the highest protection services. In addition to tending EFN, C. mimosae tend scale insects for carbohydrate-rich honeydew. We investigated the role of scale insects in this specialized ant-plant interaction. Specifically, does this putatively redundant third partner strengthen the ant-plant mutualism by making the ant a better protector of the tree? Or does it weaken the mutualism by being costly to the tree while providing no additional benefit to the ant-plant mutualism? We coupled observational surveys with two scale-manipulation experiments and found evidence that this third partner strengthens the ant-plant mutualism. Trees with scale insects experimentally removed experienced a 2.5X increase in elephant damage compared to trees with scale insects present over 10 months. Reduced protection was driven by scale removal causing a decrease in ant colony size and per capita baseline activity and defensive behavior. We also found that ants increased scale-tending and the density of scale insects on trees when EFN were experimentally reduced. Thus, in this system, scale insects and EFN are likely complementary, rather than redundant, resources with scale insects benefitting ants when EFN production is low (such as during annual dry periods in this semi-arid ecosystem). This study reveals that a third-partner strengthens an ant-plant mutualism that serves to stabilize a whole ecosystem.

Exploring fungus-plant N transfer in a tripartite ant-plant-fungus mutualism.

Background and Aims: The plant Hirtella physophora, the ant Allomerus decemarticulatus and a fungus, Trimmatostroma sp., form a tripartite association. The ants manipulate both the plant trichomes and the fungus to build galleries under the stems of their host plant used to capture prey. In addition to its structural role, the fungus also improves nutrient uptake by the host plant. But it still remains unclear whether the fungus plays an indirect or a direct role in transferring nutrients to the plant. This study aimed to trace the transfer of N from the fungus to the plant's stem tissue. Methods: Optical microscopy and transmission electron microscopy (TEM) were used to investigate the presence of fungal hyphae in the stem tissues. Then, a 15N-labelling experiment was combined with a nanoscale secondary-ion mass spectrometry (NanoSIMS 50) isotopic imaging approach to trace the movement of added 15N from the fungus to plant tissues. Key Results: The TEM images clearly showed hyphae inside the stem tissue in the cellular compartment. Also, fungal hyphae were seen perforating the wall of the parenchyma cell. The 15N provisioning of the fungus in the galleries resulted in significant enrichment of the 15N signature of the plant's leaves 1 d after the 15N-labelling solution was deposited on the fungus-bearing trap. Finally, NanoSIMS imaging proved that nitrogen was transferred biotrophically from the fungus to the stem tissue. Conclusions: This study provides evidence that the fungi are connected endophytically to an ant-plant system and actively transfer nitrogen from 15N-labelling solution to the plant's stem tissues. Overall, this study underlines how complex the trophic structure of ant-plant interactions is due to the presence of the fungus and provides insight into the possibly important nutritional aspects and tradeoffs involved in myrmecophyte-ant mutualisms.

Network reorganization and breakdown of an ant-plant protection mutualism with elevation.

Both the abiotic environment and the composition of animal and plant communities change with elevation. For mutualistic species, these changes are expected to result in altered partner availability, and shifts in context-dependent benefits for partners. To test these predictions, we assessed the network structure of terrestrial ant-plant mutualists and how the benefits to plants of ant inhabitation changed with elevation in tropical forest in Papua New Guinea. At higher elevations, ant-plants were rarer, species richness of both ants and plants decreased, and the average ant or plant species interacted with fewer partners. However, networks became increasingly connected and less specialized, more than could be accounted for by reductions in ant-plant abundance. On the most common ant-plant, ants recruited less and spent less time attacking a surrogate herbivore at higher elevations, and herbivory damage increased. These changes were driven by turnover of ant species rather than by within-species shifts in protective behaviour. We speculate that reduced partner availability at higher elevations results in less specialized networks, while lower temperatures mean that even for ant-inhabited plants, benefits are reduced. Under increased abiotic stress, mutualistic networks can break down, owing to a combination of lower population sizes, and a reduction in context-dependent mutualistic benefits.

Community analysis of microbial sharing and specialization in a Costa Rican ant-plant-hemipteran symbiosis.

Ants have long been renowned for their intimate mutualisms with trophobionts and plants and more recently appreciated for their widespread and diverse interactions with microbes. An open question in symbiosis research is the extent to which environmental influence, including the exchange of microbes between interacting macroorganisms, affects the composition and function of symbiotic microbial communities. Here we approached this question by investigating symbiosis within symbiosis. Ant-plant-hemipteran symbioses are hallmarks of tropical ecosystems that produce persistent close contact among the macroorganism partners, which then have substantial opportunity to exchange symbiotic microbes. We used metabarcoding and quantitative PCR to examine community structure of both bacteria and fungi in a Neotropical ant-plant-scale-insect symbiosis. Both phloem-feeding scale insects and honeydew-feeding ants make use of microbial symbionts to subsist on phloem-derived diets of suboptimal nutritional quality. Among the insects examined here, Cephalotes ants and pseudococcid scale insects had the most specialized bacterial symbionts, whereas Azteca ants appeared to consume or associate with more fungi than bacteria, and coccid scale insects were associated with unusually diverse bacterial communities. Despite these differences, we also identified apparent sharing of microbes among the macro-partners. How microbial exchanges affect the consumer-resource interactions that shape the evolution of ant-plant-hemipteran symbioses is an exciting question that awaits further research.

Limited gene dispersal and spatial genetic structure as stabilizing factors in an ant-plant mutualism.

Comparative studies of the population genetics of closely associated species are necessary to properly understand the evolution of these relationships because gene flow between populations affects the partners' evolutionary potential at the local scale. As a consequence (at least for antagonistic interactions), asymmetries in the strength of the genetic structures of the partner populations can result in one partner having a co-evolutionary advantage. Here, we assess the population genetic structure of partners engaged in a species-specific and obligatory mutualism: the Neotropical ant-plant, Hirtella physophora, and its ant associate, Allomerus decemarticulatus. Although the ant cannot complete its life cycle elsewhere than on H. physophora and the plant cannot live for long without the protection provided by A. decemarticulatus, these species also have antagonistic interactions: the ants have been shown to benefit from castrating their host plant and the plant is able to retaliate against too virulent ant colonies. We found similar short dispersal distances for both partners, resulting in the local transmission of the association and, thus, inbred populations in which too virulent castrating ants face the risk of local extinction due to the absence of H. physophora offspring. On the other hand, we show that the plant populations probably experienced greater gene flow than did the ant populations, thus enhancing the evolutionary potential of the plants. We conclude that such levels of spatial structure in the partners' populations can increase the stability of the mutualistic relationship. Indeed, the local transmission of the association enables partial alignments of the partners' interests, and population connectivity allows the plant retaliation mechanisms to be locally adapted to the castration behaviour of their symbionts.

Cryptic coloration of Macaranga bancana seedlings: A unique strategy for a pioneer species.

Macaranga bancana is considered as a successful pioneer plant species. Usually found in disturbed and open areas, most of the current research focused on its relations with ants. One of the unique feature of the plants is that the seedling leaves are red, resembling and almost matching the background. Using a portable spectrometer, we measured the color reflectance of M. bancana seedlings (less than 20 cm in height). We also measured the leaf litter reflectance, adult M. bancana leaves and also seedlings of several other species found in the vicinity of M. bancana seedlings. The reflectances of M. bancana seedlings are very similar to that of the leaf litter background. We suggest that this cryptic coloration is crucial during the early stages of the plant when it still cannot rely on the protection of ants.

Morphological and phylogenetic investigations for several cryptic ant-plants found in Callicarpa (Lamiaceae) from Borneo.

A tropical small tree, Callicarpa saccata, is known to have a symbiotic relationship with ants. It has sac-like structures at the base of the leaves that are inhabited by ants. No other species has been determined to be a myrmecophyte among the ca. 140 species of this genus. However, our recent field investigation discovered that two other species on Borneo (C. barbata and C. teneriflora) have hollow stems, which seem to be inhabited by ants. We observed the morphological features of these species in relation to their usage by ants, and became convinced that they are mymecophytic species. The molecular phylogenetic analyses using ITS and chloroplast regions suggest that C. saccata and C. teneriflora are closely related, but the differences in the myrmecophytic features of these species should be noted.

Current issues in the evolutionary ecology of ant-plant symbioses.

Ant-plant symbioses involve plants that provide hollow structures specialized for housing ants and often food to ants. In return, the inhabiting ants protect plants against herbivores and sometimes provide them with nutrients. Here, we review recent advances in ant-plant symbioses, focusing on three areas. First, the nutritional ecology of plant-ants, which is based not only on plant-derived food rewards, but also on inputs from other symbiotic partners, in particular fungi and possibly bacteria. Food and protection are the most important 'currencies' exchanged between partners and they drive the nature and evolution of the relationships. Secondly, studies of conflict and cooperation in ant-plant symbioses have contributed key insights into the evolution and maintenance of mutualism, particularly how partner-mediated feedbacks affect the specificity and stability of mutualisms. There is little evidence that mutualistic ants or plants are under selection to cheat, but the costs and benefits of ant-plant interactions do vary with environmental factors, making them vulnerable to natural or anthropogenic environmental change. Thus, thirdly, ant-plant symbioses should be considered good models for investigating the effects of global change on the outcome of mutualistic interactions.

Effects of dam-induced landscape fragmentation on amazonian ant-plant mutualistic networks.

Mutualistic networks are critical to biological diversity maintenance; however, their structures and functionality may be threatened by a swiftly changing world. In the Amazon, the increasing number of dams poses a large threat to biological diversity because they greatly alter and fragment the surrounding landscape. Tight coevolutionary interactions typical of tropical forests, such as the ant-myrmecophyte mutualism, where the myrmecophyte plants provide domatia nesting space to their symbiotic ants, may be jeopardized by the landscape changes caused by dams. We analyzed 31 ant-myrmecophyte mutualistic networks in undisturbed and disturbed sites surrounding Balbina, the largest Central Amazonian dam. We tested how ant-myrmecophyte networks differ among dam-induced islands, lake edges, and undisturbed forests in terms of species richness, composition, structure, and robustness (number of species remaining in the network after partner extinctions). We also tested how landscape configuration in terms of area, isolation, shape, and neighborhood alters the structure of the ant-myrmecophyte networks on islands. Ant-myrmecophytic networks were highly compartmentalized in undisturbed forests, and the compartments had few strongly connected mutualistic partners. In contrast, networks at lake edges and on islands were not compartmentalized and were negatively affected by island area and isolation in terms of species richness, density, and composition. Habitat loss and fragmentation led to coextinction cascades that contributed to the elimination of entire ant-plant compartments. Furthermore, many myrmecophytic plants in disturbed sites lost their mutualistic ant partners or were colonized by opportunistic, nonspecialized ants. Robustness of ant-myrmecophyte networks on islands was lower than robustness near lake edges and in undisturbed forest and was particularly susceptible to the extinction of plants. Beyond the immediate habitat loss caused by the building of large dams in Amazonia, persistent edge effects and habitat fragmentation associated with dams had large negative effects on animal-plant mutualistic networks.

Relação entre diferentes espécies de formigas e a mirmecófita Cordia nodosa Lamarck (Boraginaceae) em áreas de mata ripária na Amazônia mato-grossense / Relationship among different plant-ants and its myrmecophite host Cordia nodosa Lamarck (Boraginaceae) in a riparian Mato Grosso Amazonian forest

Os benefícios obtidos por um organismo em uma associação mutualística podem variar em função de fatores ambientais, bem como entre as diferentes espécies que podem estar associadas. Neste trabalho demonstramos que quatro espécies de formigas, Crematogaster brasiliensis, Allomerus octoarticulatus e duas não identificadas do gênero Azteca podem ser encontradas associadas à mirmecófita Cordia nodosa em florestas ripárias sul-amazônicas. Essa composição de espécies de formigas é mais similar a encontrada na Amazônia Andina do que aquela da Amazônia Central brasileira. A colonização por formigas parece ser determinante, pois diminuiu a herbivoria e, consequentemente, aumentou a probabilidade de C. nodosa produzir frutos. Adicionalmente, mesmo não havendo diferença na herbivoria entre plantas colonizadas pelas diferentes espécies de formigas, a probabilidade de uma planta colonizada por formigas do gênero Allomerus produzir frutos é menor do que quando colonizadas pelas outras espécies de formigas. Esse estudo demonstra a dependência de C. nodosa pela colonização de formigas para sua reprodução. Contudo, conforme outros estudos realizados em outras áreas da Amazônia demonstram, nossos resultados também sugerem que Allomerus pode estar castrando as plantas hospedeiras, agindo como parasita em toda a sua distribuição geográfica.

The benefits obtained by an organism when involved in a mutualistic interaction vary depending on environmental factors, as well as among the identity of the involved species. In this study, we showed that four ant species, Crematogaster brasiliensis, Allomerus octoarticulatus, and two unidentified Azteca species can be found associated to the myrmecophite Cordia nodosa in riparian forests in the South of Amazonia. This composition of ant-associated species is more similar in forests of Andean Amazon than in Central Amazonia. The colonization of an ant colony on C. nodosa seems to be vital in order to decrease herbivory, as increased the probability of a plant sets fruits. Moreover, even though we did not find significant differences in herbivory among plants colonized by different ant species, the probability of a plant produces fruits is much lower when it is colonized by Allomerus ants. Overall, this study shows that C. nodosa depends on ants to reproduce. However, based on other empirical studies across the Amazon, our results also suggest that Allomerus ants can act as flower castrator, acting as a parasite over its geographic range.

Ant-plant associations in different forests in Venezuela / Associações formiga-planta em diferentes florestas na Venezuela

We evaluated the hypothesis that the abundance and species distribution of two different kinds of myrmecophilous plants is influenced differently by the ant diversity and abundance. In eight different natural forests in Venezuela we estimated the species richness and abundance of plants, ants on the soil and on the canopy, the leaf damage of plants and soil nutrients. The main results of the study show that plants with domatia (PD) and plants with extrafloral nectaries (PEFN) have different relationships with ants and suffer from different ecological constraints. PD attract a more specific group of ants than PEFN. Our results are consistent with the hypothesis that domatia are adaptations that help plants to increase rare nutrient uptake rather than for herbivore defense. We found that PEFN attract a larger variety of ant species than PD, and ant abundance seems to limit the ecological range of PEFN. The attraction of ants as a mechanism to reduce herbivory, as done by PEFN, does not seem to be superior to alternative anti-herbivore mechanisms used by other plants. Contrary to many former studies, we found that ants are generally more diverse on the soil compared to canopies.

Avaliou-se a hipótese de que a distribuição da abundância e o número de espécie de dois tipos diferentes de plantas mirmecófilas são influenciadas diferentemente pela diversidade e pela abundância de formigas. Em oito florestas naturais diferentes na Venezuela nós estimamos a riqueza de espécies e a abundância de plantas, de formigas no solo e no dossel, os danos às folhas e os nutrientes do solo. Os resultados mostram que as plantas com domácias para alojar formigas e plantas com nectários extraflorais (PEFN) têm relacionamentos diferentes com formigas e sofrem confinamentos ecológicos diferentes. As plantas com domácias atraem um grupo mais específico de formigas do que as PEFN. Os resultados são consistentes com a hipótese de que as domácias são adaptações que ajudam a planta a aumentar o acesso a nutrientes escassos para a defesa de herbívoros. PEFN atraem uma variedade maior da espécie de formigas do que as plantas com domácias, e a abundância de formigas parece limitar a escala ecológica de PEFN. A atração das formigas como mecanismo para reduzir a herbivoria, como feito por PEFN, não parece ser superior aos mecanismos alternativos de prevenção de herbivoria usados por outras plantas. Contrário a muitos estudos anteriores, as formigas mostraram-se geralmente mais diversas no solo comparado ao dossel.

Plants feed ants: food bodies of myrmecophytic Piper and their significance for the interaction with Pheidole bicornis ants.

Several species of Piper (Piperaceae) live in symbiosis with Pheidole bicornis (Formicidae-Myrmicinae) on the southern Pacific slope of Costa Rica. These plants produce small single-celled food bodies (FBs) in leaf domatia, formed by the petiole bases and roofing leaf sheaths. In the present study the dependency of ants on FBs of Piper fimbriulatum as a food source was analysed by comparing the natural abundance of 13C and 15N in ants and FBs. Both δ13C and δ15N values were very similar between FBs and Pheidole bicornis ants but differed substantially between the plant and other ant species. Therefore we suggest that FBs are a main food source for Pheidole bicornis ants. To strengthen this suggestion, the chemical composition of FBs of four myrmecophytic Piper species was analysed, with special emphasis on the nutritional requirements of inhabiting Pheidole bicornis ants. Standard chemical methods were modified and combined to a novel analysis scheme by which all major FB constituents could be quantified from minute [3-10 mg dry mass (DM)] quantities. Piper FBs mainly consisted of lipids (41-48% of DM) and proteins (17-24% of DM). Soluble carbohydrates and amino acids proved to be quantitatively unimportant. N was predominantly stored as soluble protein and, thus, was easily available to the ants. FBs proved to be a high-energy food source (up to 23 kJ g-1 DM), with a chemical composition that meets well the nutritional needs of the inhabiting ants.