Ants,plant characteristics and habitat conservation status affect the occurrence of myrmecophilous butterflies on an extrafloral nectaried Malpighiaceae

We investigated the occurrence of myrmecophilous florivorous lycaenid larvae in Banisteriopsis malifolia (Malpighiaceae) according to habitat conservation status (disturbed and preserved savanna), plant phenology, height and the presence of tending ants. The abundance and richness of lycaenids were sixfold and fivefold greater, respectively, in the disturbed area than in the protected savanna. Lycaenids occurred mostly on plants visited by Camponotus blandus, a mutualistic partner of larvae. Habitat type was the main factor influencing lycaenid occurrence, as plants in open areas offered more food resources and tending ants. Banisteriopsis malifolia was considered useful as a host for lycaenids in disturbed sites.     

Elevated temperatures alter an ant‐aphid mutualism

Ant‐hemipteran mutualisms are keystone interactions that can be variously affected by warming: these mutualisms can be strengthened or weakened, or the species can transition to new mutualist partners. We examined the effects of elevated temperatures on an ant‐aphid mutualism in the subalpine zone of the Rocky Mountains in Colorado, USA. In this system, inflorescences of the host plant, Ligusticum porteri Coult. & Rose (Apiaceae), are colonized by the ant‐tended aphid Aphis asclepiadis Fitch or less frequently by the non‐ant tended aphid Cavariella aegopodii (Scopoli) (both Hemiptera: Aphididae). Using an 8‐year observational study, we tested for two key mechanisms by which ant‐hemipteran mutualisms may be altered by climate change: shifts in species identity and phenological mismatch. Whereas the aphid species colonizing the host plant is not changing in response to year‐to‐year variation in temperature, we found evidence that a phenological mismatch between ants and aphids could occur. In warmer years, colonization of host plant inflorescences by ants is decreased, whereas for A. asclepiadis aphids, host plant colonization is mostly responsive to date of snowmelt. We also experimentally established A. asclepiadis colonies on replicate host plants at ambient and elevated temperatures. Ant abundance did not differ between aphid colonies at ambient vs. elevated temperatures, but ants were less likely to engage in tending behaviors on aphid colonies at elevated temperatures. Sugar composition of aphid honeydew was also altered by experimental warming. Despite reduced tending by ants, aphid colonies at elevated temperatures had fewer intraguild predators. Altogether, our results suggest that higher temperatures may disrupt this ant‐aphid mutualism through both phenological mismatch and by altering benefits exchanged in the interaction.     

Orchid seed removal by ants in Neotropical ant‐gardens

• Most plants that inhabit ant‐gardens (AGs) are cultivated by the ants. Some orchids occur in AGs; however, it is not known whether their seeds are dispersed by AG ants because most orchid seeds are tiny and dispersed by wind.
• We performed in situ seed removal experiments, in which we simultaneously provided Azteca gnava ants with seeds of three AG orchid species and three other AG epiphyte species (Bromeliaceae, Cactaceae and Gesneriaceae), as well as the non‐AG orchid Catasetum integerrimum.
• The seeds most removed were those of the bromeliad Aechmea tillandsioides and the gesneriad Codonanthe uleana, while seeds of AG orchids Coryanthes picturata, Epidendrum flexuosum and Epidendrum pachyrachis were less removed. The non‐AG orchid was not removed. Removal values were positively correlated with the frequency of the AG epiphytes in the AGs, and seeds of AG orchids were larger than those of non‐AG orchids, which should favour myrmecochory.
• Our data show that Azt. gnava ants discriminate and preferentially remove seeds of the AG epiphytes. We report for the first time the removal of AG orchid seeds by AG ants in Neotropical AGs.

     

Quarantining behaviour of ants towards infected aphids as an antifungal mechanism in ant–aphid interactions

The ecological success of social insects, including ants, is tightly connected with their ability to protect themselves and their food resources. In exchange for energy‐rich honeydew, ants protect myrmecophilous aphids from various natural enemies. Fungal infection can have disastrous consequences for both mutualist partners, wherein aphids can be disease vectors. Behavioural responses towards fungus‐infected aphids of ant species in nature have scarcely been studied. Here, we studied the behaviour of honeydew foragers of four ant species – Formica polyctena Foerster, Formica rufa L., Formica pratensis Retzius (Hymenoptera: Formicidae, Formicini), and Lasius niger (L.) (Formicidae, Lasiini) – towards Symydobius oblongus (von Heyden) aphids contaminated with the generalist fungal pathogen Beauveria bassiana (Balsamo‐Crivelli) Vuillemin in the field. Aphid milkers from Formica spp. quickly detected and removed infected aphids from the host plant (Betula pendula Roth., Betulaceae). Neither ant species, the degree of aphid‐milker specialization (medium or high), nor the number of honeydew foragers had significant effects on the behaviour of Formica milkers towards infected aphids. Unlike Formica ants, L. niger usually displayed non‐aggressive behaviour (tolerance, antennation, honeydew collection, grooming). By the immediate removal of infected insects, Formica ants seem to minimize the probability of infection of symbionts as well as themselves. Quarantining behaviour may play an important role in ant–aphid interactions as a preventive antifungal mechanism formed under parasite pressure and thus contributing to the ecological success of ants.     

Extranuptial nectaries in flowers: ants increase the reproductive success of the ant‐plant Miconia tococa (Melastomataceae)

• Although the production of extranuptial nectar is a common strategy of indirect defence against herbivores among tropical plants, the presence of extranuptial nectaries in reproductive structures is rare, especially in ant‐plants. This is because the presence of ants in reproductive organs can generate conflicts between the partners, as ants can inhibit the activity of pollinators or even castrate their host plants. Here we evaluate the hypothesis that the ant‐plant Miconia tococa produces nectar in its petals which attracts ants and affects fruit set.
• Floral buds were analysed using anatomical and histochemical techniques. The frequency and behaviour of floral visitors were recorded in field observations. Finally, an ant exclusion experiment was conducted to evaluate the effect of ant presence on fruit production.
• The petals of M. tococa have a secretory epidermis that produces sugary compounds. Nectar production occurred during the floral bud stage and attracted 17 species of non‐obligate ants (i.e. have a facultative association with ant‐plants). Ants foraged only on floral buds, and thus did not affect the activity of pollinators in the neighbouring open flowers. The presence of ants in the inflorescences increased fruit production by 15%.
• To our knowledge, the production of extranuptial nectar in the reproductive structures of a myrmecophyte is very rare, with few records in the literature. Although studies show conflicts between the partners in the ant–plant interaction, ants that forage on M. tococa floral buds protect the plant against floral herbivores without affecting bee pollination.

     

Induced indirect defence in a spider–plant system mediated by pericarpial nectaries

Some plant species attacked by herbivore species produce additional resources to attract predators and induce an indirect defence process. We evaluated whether Palicourea rigida (Rubiaceae) individuals can induce indirect defences as response to herbivory simulation by increasing pericarpial nectar production and volatile emissions, as well as whether spiders are attracted by such induced indirect defences. We selected 30 P. rigida individuals and simulated herbivory in 15 of them by cutting out half of all leaves using pruning shears. We did not manipulate the other 15 plants (control group). At three different times, we measured nectar volume and calories of the pericarpial nectary in the inflorescences of all plants of control and treatment groups. We also quantified spider abundance on these plants. In another experiment, we selected salticid spider, Thiodina sp., to determine whether predators detect chemical tracks of plant volatiles produced by the plant after herbivory simulations. We also tested whether the honey solution could emit olfactory signals capable of attractive spiders. We showed that P. rigida produced higher volume of pericarpial nectar presenting more calories after herbivory simulation. The abundance of spiders was higher in plants subjected to herbivory simulation than control plants. Thiodina sp. did not respond to the volatile chemical tracks produced by the leaves after the simulation, but it had a positive response to olfactory tracks associated with the sucrose solution. Such an outcome indicates the ability of this spider to locate nectar honey plants and olfactory signals of honey. Thus, plants respond to the action of herbivores by producing more pericarpial nectar and nectar with more calories. Although our knowledge about the olfactory physiology of arachnids remains incipient, we highlight the importance of chemical and olfactory tracks for decision‐making of spiders in foraging on plants and the herbivory influence on the behaviour of cursorial spiders.     

Herbivory in a spider through exploitation of an ant–plant mutualism

Spiders are thought to be strict predators [1]. We describe a novel exception: Bagheera kiplingi, a Neotropical jumping spider (Salticidae) that exploits a well-studied ant–plant mutualism, is predominantly herbivorous. From behavioral field observations and stable-isotope analyses, we show that the main diet of this host-specific spider comprises specialized leaf tips (Beltian food bodies; Figure 1A) from Vachellia spp. ant-acacias (formerly Acacia spp.), structures traded for protection in the plant's coevolved mutualism with Pseudomyrmex spp. ants that inhabit its hollow thorns [2]. This is the first report of a spider that feeds primarily and deliberately on plants.     

Introduced ants reduce interaction diversity in a multi‐species,ant–aphid mutualism

Mutualisms contribute in fundamental ways to the origin, maintenance and organization of biological diversity. Introduced species commonly participate in mutualisms, but how this phenomenon affects patterns of interactions among native mutualists remains incompletely understood. Here we examine how networks of interactions among aphid‐tending ants, ant‐tended aphids, and aphid‐attacking parasitoid wasps differ between 12 spatially paired riparian study sites with and without the introduced Argentine ant Linepithema humile in southern California. To resolve challenges in species identification, we used DNA barcoding to identify aphids and screen for parasitoid wasps (developing inside their aphid hosts) from 170 aphid aggregations sampled on arroyo willow Salix lasiolepis. Compared to uninvaded sites, invaded sites supported significantly fewer species of aphid‐tending ants and ant‐tended aphids. At invaded sites, for example, we found only two species of ant‐tended aphids, which were exclusively tended by L. humile, whereas at uninvaded sites we found 20 unique ant–aphid interactions involving eight species of ant‐tended aphids and nine species of aphid‐tending ants. Ant–aphid linkage density was thus significantly lower at invaded sites compared to uninvaded sites. We detected aphid parasitoids in 14% (28/198) of all aphid aggregations. Although the level of parasitism did not differ between invaded and uninvaded sites, more species of wasps were detected within uninvaded sites compared to invaded sites. These results provide a striking example of how the assimilation of introduced species into multi‐species mutualisms can reduce interaction diversity with potential consequences for species persistence.     

A single lycaenid caterpillar gets an ant‐constructed shelter and uninterrupted ant attendance

Ant‐lycaenid associations range from mutualism to parasitism and the caterpillars of some species of lycaenids are reported to enter ant nests for shelter, diapause, or pupation. The present study aimed to examine the nature of the association between Euchrysops cnejus (Fabricius) (Lepidoptera: Lycaenidae) and Camponotus compressus (Fabricius) (Hymenoptera: Formicidae) worker ants on the extrafloral nectary‐bearing cowpea plant, Vigna unguiculata (L.) Walp. (Fabaceae). The abundance patterns of the ants and the lycaenid caterpillars together with the spatial patrolling patterns of the ants on the plants revealed that ant abundance increased with the occurrence of the lycaenid caterpillars and the ants preferred the lycaenids over the extrafloral nectar. Camponotus compressus worker ants constructed a shelter at the cowpea plant base after interaction with one or more lycaenid caterpillar(s) and tended the caterpillars and pupae till the emergence of the butterfly. The ant‐constructed shelters (ACSs) inhabited by the minor caste workers (13 ± 1.3 ants per ACS), were utilized by the caterpillars to undergo pupation. The ants confined their activities predominantly to tending the pod‐feeding caterpillars and the solitary pupa within each ACS. It appears that the behavior of the tending worker ants is modulated by the lycaenid vulnerable stages.     

Convergence,constraint and the potential for mutualism between ants and gut microbes

Ants are a hugely diverse family of eusocial insects that dominate terrestrial ecosystems all over the planet. Did mutualistic gut microbes help ants to achieve their diversity and ecological dominance? Initial studies suggested the potential for widespread convergence in ant gut bacterial communities based on dietary niche, but it now seems possible that dedicated bacterial symbionts are restricted to a minority of ant lineages (Russell et al., 2017). Nevertheless, as most ants are omnivores, the evidence so far has suggested a broad, positive correlation between the evolution of dietary specialization and ant investment in nutrient‐provisioning gut bacteria. In this issue of Molecular Ecology, Sapountzis et al. (2019) and Rubin et al. (2019) examine the evolution of gut bacterial communities in two iconic ant taxa—the attine fungus farmers and the Pseudomyrmex plant bodyguards, respectively—in a comparative context. By comparing gut bacteria between ant species of differing dietary specialization within each taxon, these studies demonstrate a hint of convergence in the midst of widespread apparent constraints. These results raise numerous interesting questions about the nature of these apparent constraints and whether they are causes or consequences of varying investment by ants to mutualism with their gut microbes.     

Polygyny does not explain the superior competitive ability of dominant ant associates in the African ant‐plant,Acacia (Vachellia) drepanolobium

The Acacia drepanolobium (also known as Vachellia drepanolobium) ant‐plant symbiosis is considered a classic case of species coexistence, in which four species of tree‐defending ants compete for nesting space in a single host tree species. Coexistence in this system has been explained by trade‐offs in the ability of the ant associates to compete with each other for occupied trees versus the ability to colonize unoccupied trees. We seek to understand the proximal reasons for how and why the ant species vary in competitive or colonizing abilities, which are largely unknown. In this study, we use RADseq‐derived SNPs to identify relatedness of workers in colonies to test the hypothesis that competitively dominant ants reach large colony sizes due to polygyny, that is, the presence of multiple egg‐laying queens in a single colony. We find that variation in polygyny is not associated with competitive ability; in fact, the most dominant species, unexpectedly, showed little evidence of polygyny. We also use these markers to investigate variation in mating behavior among the ant species and find that different species vary in the number of males fathering the offspring of each colony. Finally, we show that the nature of polygyny varies between the two commonly polygynous species, Crematogaster mimosae and Tetraponera penzigi: in C. mimosae, queens in the same colony are often related, while this is not the case for T. penzigi. These results shed light on factors influencing the evolution of species coexistence in an ant‐plant mutualism, as well as demonstrating the effectiveness of RADseq‐derived SNPs for parentage analysis.     

Reversal of mutualism in a leafflower–leafflower moth association: the possible driving role of a third‐party partner

A major goal in the study of mutualism is to understand how co‐operation is maintained when mutualism may potentially turn into parasitism. Although certain mechanisms facilitate the persistence of mutualism, parasitic species have repeatedly evolved from mutualistic ancestors. However, documented examples of mutualism reversals are still rare. Leafflowers (Phyllantheae; Phyllanthaceae) include approximately 500 species that engage in obligate mutualism with leafflower moths (Epicephala; Gracillariidae), which actively pollinate flowers, and whose larvae feed on the resulting seeds. We found that the Taiwanese population of the Phyllanthus reticulatus species complex was associated with six sympatric Epicephala species, of which three were derived parasites that induced gall formation on flowers/buds and produced no seeds. Notably, two parasitic species have retained mutualistic pollination behaviour, suggesting that the parasitism was likely not selected for to reduce the cost of mutualism. We propose that the galling habit evolved as an adaptation to escape parasitism by a specialized braconid wasp. The tough gall produced by one species was almost free of braconid parasitism, and the swollen gall induced by the other species probably prevents attack as a result of the larger airspace inside the gall. Our findings suggest that the presence of a third‐party partner can greatly influence the evolutionary fate of mutualisms, regardless of whether the pairwise interaction continues to favour co‐operation.     

Infiltration of a facultative ant–plant mutualism by the introduced Argentine ant: effects on mutualist diversity and mutualism benefits

1. Ant–plant mutualisms have been the focus of considerable empirical research, but few studies have investigated how introduced ants affect these interactions. Using 2 years of survey data, this study examines how the introduced Argentine ant [Linepithema humile (Mayr)] differs from native ants with respect to its ability to protect the extrafloral nectary‐bearing coast barrel cactus (Ferocactus viridescens) in Southern California. 2. Eighteen native ant species visited cacti in uninvaded areas, but cacti in invaded areas were primarily visited by the Argentine ant. The main herbivore of the coast barrel cactus present at the study sites is a leaf‐footed bug (Narnia wilsoni). 3. Herbivore presence (the fraction of surveys in which leaf‐footed bugs were present on individual cacti) was negatively related to ant presence (the fraction of surveys in which ants were present on individual cacti). Compared with cacti in uninvaded areas, those in invaded areas were less likely to have herbivores and when they did had them less often. 4. Seed mass was negatively related to herbivore presence, and this relationship did not differ for cacti in invaded areas versus those in uninvaded areas. 5. Although the Argentine ant might provide superior protection from herbivores, invasion‐induced reductions in ant mutualist diversity could potentially compromise plant reproduction. The cumulative number of ant species on individual cacti over time was lower in invaded areas and was associated with a shortened seasonal duration of ant protection and reduced seed mass. These results support the hypothesis that multiple partners may enhance mutualism benefits.     

Genetic and evolution analysis of extrafloral nectary in cotton

Extrafloral nectaries are a defence trait that plays important roles in plant–animal interactions. Gossypium species are characterized by cellular grooves in leaf midribs that secret large amounts of nectar. Here, with a panel of 215 G. arboreum accessions, we compared extrafloral nectaries to nectariless accessions to identify a region of Chr12 that showed strong differentiation and overlapped with signals from GWAS of nectaries. Fine mapping of an F₂ population identified GaNEC1, encoding a PB1 domain‐containing protein, as a positive regulator of nectary formation. An InDel, encoding a five amino acid deletion, together with a nonsynonymous substitution, was predicted to cause 3D structural changes in GaNEC1 protein that could confer the nectariless phenotype. mRNA‐Seq analysis showed that JA‐related genes are up‐regulated and cell wall‐related genes are down‐regulated in the nectary. Silencing of GaNEC1 led to a smaller size of foliar nectary phenotype. Metabolomics analysis identified more than 400 metabolites in nectar, including expected saccharides and amino acids. The identification of GaNEC1 helps establish the network regulating nectary formation and nectar secretion, and has implications for understanding the production of secondary metabolites in nectar. Our results will deepen our understanding of plant–mutualism co‐evolution and interactions, and will enable utilization of a plant defence trait in cotton breeding efforts.     

Unique haplotypes in ant‐attended aphids and widespread haplotypes in non‐attended aphids

Aphid species within the genus Tuberculatus Mordvilko (Hemiptera: Aphididae) exhibit a variety of interactions with ants, ranging from close associations to non‐attendance. A previous study indicated that despite wing possession, ant‐attended Tuberculatus species exhibited low dispersal rates compared with non‐attended species. This study examined if presence or absence of mutualistic interactions and habitat continuity of host plants affected intraspecific genetic diversity and genetic differentiation in mitochondrial DNA cytochrome oxidase I (COI) sequences. Sympatric ant‐attended Tuberculatus quercicola (Matsumura) (Hemiptera: Aphididae) and non‐attended Tuberculatus paiki Hille Ris Lambers (Hemiptera: Aphididae) were collected from the daimyo oak Quercus dentata Thunberg (Fagales: Fagaceae) in Japan and examined for haplotype variability. Seventeen haplotypes were identified in 568 T. quercicola individuals representing 23 populations and seven haplotypes in 425 T. paiki representing 19 populations. Haplotype diversity, which indicates the mean number of differences between all pairs of haplotypes in the sample, and nucleotide diversity were higher in T. quercicola than T. paiki. Analysis of molecular variance (AMOVA) showed higher genetic differentiation among populations within groups of T. quercicola (39.8%) than T. paiki (22.6%). The effects of attendant ant species on genetic differentiation in T. quercicola were not distinguishable from geographic factors. Despite low dispersal rates, host plant habitat continuity might facilitate widespread dispersal of a T. quercicola haplotype in Hokkaido. These results suggested that following T. quercicola colonization, gene flow among populations was limited, resulting in genetic drift within populations. However, frequent T. paiki dispersal is clearly evident by low genetic differentiation among populations within groups, resulting in lower haplotype diversity.     

Day vs. night predation on artificial caterpillars in primary rainforest habitats – an experimental approach

The influence of natural enemies has led to the evolution of various predator avoidance strategies in herbivorous insects. Many caterpillars are exclusively active at night and rest during the day. It is widely assumed that nocturnal activity in caterpillars reduces their risk of falling prey to their natural enemies. To test this hypothesis, we compared predation pressure between day and night in tree‐fall gaps and closed‐canopy forest sites in an Amazonian primary lowland rainforest. Artificial clay caterpillars, showing camouflaged colouration (green), were exposed as potential prey to a natural predator community. Attacks were significantly more frequent during daytime and were reduced by about a quarter at night in tree‐fall gaps, and by a third in closed‐canopy forest sites. This supports the idea of time‐dependent activity in caterpillars as an antipredatory adaptation. Further, independent of the time of day, predation pressure on caterpillars was significantly higher in tree‐fall gaps compared to closed‐canopy forest habitats. Nearly all predation events were caused by arthropods, whereas birds played a negligible role. Across both habitat types and time scales, ants acted as major predator group, emphasising their important role in population control of herbivorous insects in lowland rainforest ecosystems. This is the first experimental study using artificial caterpillars to examine whether time‐scheduling of exposition might influence predation risk amongst undefended, solitary, free‐living lepidopteran larvae.     

Photosynthesis and isoprene emission from trees along an urban–rural gradient in Texas

Isoprene emission is an important mechanism for improving the thermotolerance of plant photosystems as temperatures increase. In this study, we measured photosynthesis and isoprene emission in trees along an urban–rural gradient that serves as a proxy for climate change, to understand daily and seasonal responses to changes in temperature and other environmental variables. Leaf‐level gas exchange and basal isoprene emission of post oak (Quercus stellata) and sweet gum (Liquidambar styraciflua) were recorded at regular intervals over an entire growing season at urban, suburban, and rural sites in eastern Texas. In addition, the temperature and atmospheric carbon dioxide concentration experienced by leaves were experimentally manipulated in spring, early summer, and late summer. We found that trees experienced lower stomatal conductance and photosynthesis and higher isoprene emission, at the urban and suburban sites compared to the rural site. Path analysis indicated a daily positive effect of isoprene emission on photosynthesis, but unexpectedly, higher isoprene emission from urban trees was not associated with improved photosynthesis as temperatures increased during the growing season. Furthermore, urban trees experienced relatively higher isoprene emission at high CO₂ concentrations, while isoprene emission was suppressed at the other sites. These results suggest that isoprene emission may be less beneficial in urban, and potentially future, environmental conditions, particularly if higher temperatures override the suppressive effects of high CO₂ on isoprene emission. These are important considerations for modeling future biosphere–atmosphere interactions and for understanding tree physiological responses to climate change.