Thesis (Honors)--College of William and Mary, 2009. / Includes bibliographical references (leaves 52-54). Also available via the World Wide Web.

In mutualisms, individuals exchange goods and services for net benefit. However, many sources of variation in these interactions remain unexplored. To examine why plantdwelling ants protect their host plants by killing herbivores, I shifted the macronutrient balance of their background diets. Providing ants with supplemental protein caused them to invest less in patrolling and defence activities, likely because the availability of lowcost protein made hunting for herbivores relatively less profitable. In contrast, supplemental sugar caused an increase in ant activity, possibly because carbohydrates serve as ‘fuel’ for patrolling. To examine a second source of variation in this interaction, I treated ants with an activator of PKG, a protein encoded by foraging, a gene with behavioural functions in other taxa. PKG activation caused ants to become more aggressive towards herbivores, causing their host plants to experience less herbivory. This suggests that an ortholog of foraging may influence cooperation in this system.

In mutualisms, individuals exchange goods and services for net benefit. However, many sources of variation in these interactions remain unexplored. To examine why plantdwelling ants protect their host plants by killing herbivores, I shifted the macronutrient balance of their background diets. Providing ants with supplemental protein caused them to invest less in patrolling and defence activities, likely because the availability of lowcost protein made hunting for herbivores relatively less profitable. In contrast, supplemental sugar caused an increase in ant activity, possibly because carbohydrates serve as ‘fuel’ for patrolling. To examine a second source of variation in this interaction, I treated ants with an activator of PKG, a protein encoded by foraging, a gene with behavioural functions in other taxa. PKG activation caused ants to become more aggressive towards herbivores, causing their host plants to experience less herbivory. This suggests that an ortholog of foraging may influence cooperation in this system.

Thesis (M.S.)--Villanova University, 2009. / Biology Dept. Includes bibliographical references.

Thesis (M.S.)--Villanova University, 2009. / Biology Dept. Includes bibliographical references.

Fig trees (Ficus spp.) and fig wasps (Hymenoptera: Agaonidae) are partners in an intimate mutualism. The trees provide ovules in which wasp larvae develop while the wasps pollinate the flowers and are therefore indispensible for fig seed production. Agaonid fig wasps oviposit down the styles of fig flowers and it has generally been accepted that they were unable to reach the ovules of "long" styled flowers , which would produce seeds, thus maintaining an evolutionary stable mutualism. African fig species were found to have unimodal style length frequencies, with no separation into long and short styled flowers. In several species the ovipositors of their associated agaonids were long enough to reach the majority of ovules. The number of foundress agaonids entering a fig influenced fig seed set and therefore was an important factor regulating the proportion of flowers producing seeds or pollinators. In the two Ficus species that were studied, entry of more than three agaonid foundresses into one fig resulted in competition for limited oviposition sites and less female - biased offspring sex ratios. It is hypothesised that sequential laying of male eggs followed by female eggs, under variable oviposition site limitation, results in sex ratio adjustment, as predicted by local mate competition theory. Evidence in support of this hypothesis is presented. A number of non - pollinating torymid and pteromalid fig wasps also oviposit into each fig species. The sycophagines and sycoecines oviposit down the styles from inside the fig inflorescences like their agaonid counterparts, while other species insert their ovipositors through the wall of the fig from the outside. Like the agaonids, sycophagines were characterised by being pro - ovigenic, with numerous fully developed eggs at emergence. Sycoecines were able to re - emerge from figs they had oviposited in and lay their eggs in more than one fig. They had short ovipositors, allowing access to a smaller proportion of flowers than agaonids or sycophagines. Externally ovipositing fig wasps were syn-ovigenic, able to develop eggs as adults and invested more energy and time during each oviposition event. Differences in the ovipositor lengths of these species did not segregate their oviposition sites spatially, and therefore does not reduce competition between species. Attack by parasitoids and inquiline fig wasps from the exterior did not constitute a selection pressure against agaonids ovipositing in ovules closer to the periphery of the fig's surface, as predicted by Michaloud's enemy-free-space hypothes is. It cannot therefore explain the preference shown by ovipositing agaonids for shorter styled flowers.

The steps by which neutral, random and/or negative biological interactions evolve into mutualistic ones remain poorly understood. Here, we study Elacatinus gobies and the fishes from which they clean parasites, termed 'clients'. Colorful stripes are common to mutualist cleaners and non-cleaning sister species. Blue stripes are unique to obligate cleaners. We quantified the contrast potential of ancestral and novel stripe colors, using fish color vision models, and determined that color stripes have become more visible to clients over evolutionary time. In turn, we focused on the role of color as a potentially specialized signal. We show that cleaners possess a putative chemical defense (one multimedia file in .mov format included) and demonstrate that stripes are sufficient to elicit client stereotypical posing behavior and to deter attack. Analysis of previously published records show that yellow cleaners tend to predators less than expected, compared to green and blue cleaners. Our results highlight evolution from predator avoidance to tolerance with conspicuous advertising reinforced by chemical defense in cleaners. Similar trajectories, via recognizable signals to risky partners, may be common in other diffuse mutualisms. We discuss the generality of defense and signal traits in other species rich lineages of mutualists.

Acacia ants evolved obligate protective mutualisms with acacia trees, which they defend against herbivores, food parasites and encroaching vegetation. In this mutualism, the fitness of one partner entirely depends on the other. Other ant species are parasitic on acacia trees; they nest on the tree, harvest food rewards, do not defend their own tree, and occasionally try to steal food from other trees, usually inhabited by mutualistic ants. To understand the behavioral and anatomical effects of the interaction between ants and host trees, I integrated brain anatomy, morphology and field experiments to study parasitic and mutualistic species of Pseudomyrmex ants associated with acacia trees. In Chapter 1, I describe a previously unknown behavior of stealing food from other ant-defended acacia trees in the parasitic acacia ant P. nigropilosus, and I evaluate four strategies that may allow parasitic ants to overcome the usually effective defenses of the robbed mutualistic ants protecting a host tree. In Chapter 2, I study how colony size correlates with the degree of division of labor and brain anatomy of workers, focusing on a species of acacia ant lacking morphological castes among workers, P. spinicola. In Chapter 3, I study acacia-ant behavior of killing vegetation encroaching on a host tree. I document the interspecific differences among acacia ants in the size of the area around the host tree that workers clear from encroaching vegetation. I further test for interspecific variation in pruning behavior, and whether mandibular force correlate with worker pruning decisions. In Chapter 4, I test whether ant species that routinely leave the host tree to forage or to prune encroaching vegetation are better at orienting themselves when returning to their host tree, compared to ant species that rarely leave their host tree. This dissertation documents how the obligate protective mutualism of an ant with a tree has consequences for division of labor, navigational skills, behavioral specializations, head shape and brain anatomy of ant workers. / text

Dissertation (Ph.D.)--Auburn University, 2006. / Abstract. Includes bibliographic references.

Mutualistic interactions between species are balanced on a delicate scale of net benefits to both interacting partners. The dynamics of such interactions could change depending on the context in which these interactions occur. One of the most well-studied models for interspecies mutualisms are myrmecophytic systems, also known as ant-plant systems, where the host plant (myrmecophyte) provides shelter (domatia), solely or along with food resources, for ant partners, while the domatia-resident ants intensively patrol and protect the host plant from herbivory. In some cases, nutrient flux has also been reported from the ant-derived debris in the domatia to the host plant. Such mutualisms are often vulnerable to exploitation by non-mutualist organisms or interlopers such as non-protective ants and other invertebrates that use the plant rewards without any returns. Since provision of domatia and food imposes costs on the host plants, the trajectory of evolution in such cases where protection is partial or absent needs investigation to understand the evolution of myrmecophytism. In this thesis, we investigate the possibility of evolution of myrmecophytism in the absence of universal protection by partner ants, using the unique semi-myrmecophyte (domatia are not expressed in all individuals of the species) Humboldtia brunonis as the study model. H. brunonis is endemic to the tropical wet evergreen forests of the Western Ghats of India. Being locally abundant in its distribution range, this plant species has also been used in characterising forest types in the Western Ghats. H. brunonis provides domatia (modified stem internodes) and food for ants in the form of extrafloral nectar (EFN) on leaves and bracts of floral buds in all individuals. Each domatium has a self-opening slit, which could have led to the domatia being accessed and inhabited by numerous ants and other non-ant invertebrates throughout its distribution range. Of these, only one ant species, Technomyrmex albipes, has been reported to be significantly protective against herbivores, and the protection received by the plant is reported to be restricted only to one site where T. albipes is most abundant. In the light of the above, the possible explanation for continued expression of rewards (domatia as well as EFN) in the absence of universal protection was investigated. Chapter 1: Introduction This chapter starts with a brief history of the concept of mutualism, evolution and maintenance of mutualism, and trophic mutualism amongst organisms in general. This is followed by a description of ant–plant mutualisms, and the various interactions that drive the interaction in such systems, with a more detailed emphasis on trophic mutualism in ant-plants, and stable isotope analysis as a technique that is used to study trophic mutualism in ant-plants. The study system, Humboldtia brunonis is introduced, and all the studies on this system preceding this current thesis are discussed in the light of findings in other ant-plant systems. Lastly, the objectives of the thesis are briefly introduced as separate chapters. Chapter 2: Context dependency of rewards and services in an Indian ant–plant interaction: southern sites favour the mutualism between plants and ants (published in Journal of Tropical Ecology) Earlier studies on the H. brunonis system have shown that there is geographic variation in the occupancy of the domatia, with domatia in the northern part of the H. brunonis range being dominantly occupied by an arboreal earthworm species, while domatia in the south are mostly occupied by ants, especially the sole protective ant T. albipes. Further, it has been reported that herbivory is significantly reduced in the presence of ants in the south. In the present study, conducted at 5 sites spanning the distribution range of H. brunonis, we observed that there is a geographic variation in various ant-related plant traits such as abundance of domatia-bearing individuals, number of nectaries per leaf, size of nectaries, and volume and composition of the EFN, with a clear north–south increasing gradient. However, strong protection mutualism was observed only at one site in the south where herbivory pressure was highest. By comparing our results with earlier findings, we show that in addition to geographic variation, there is also temporal variation in the strength of protection mutualism, and that protection mutualism in this system is context-dependent. These results provide new perspectives on the evolution of myrmecophytism. Chapter 3: Leaf expansion and foliar extrafloral nectar as defence strategies in a paleotropical ant-plant Humboldtia brunonis (Fabaceae) (a section of this chapter is submitted to Biotropica) Despite the absence of universal protection against herbivory, H. brunonis plants constitutively secrete EFN and domatia. We therefore explored other non-chemical defences in this system, and investigated possible explanations for the continued reward production. We observed rapid rate of leaf expansion during the early and most vulnerable phase of leaf phenology, and propose this as a strategy to escape herbivory. The young leaves are also subject to being infested by phloem-feeding Hemiptera, but there was seldom any case of ants tending Hemiptera for honeydew (sugary material excreted by the Hemiptera) on the plant. We analysed the sugar and amino acids compositions of EFN, honeydew and phloem sap, and found that EFN composition was much richer and more attractive (to ants) than honeydew, thereby suggesting that EFN could possibly function to distract ants from tending Hemiptera on the plant, thereby avoiding further damage to the plant. We also observed that EFN composition was much richer than phloem sap, and thereby confirmed that EFN is not mere phloem exudate; rather, our results suggests that EFN could possibly be synthesized actively in the secretory cells of the extrafloral nectary. Anatomical observations of the foliar nectaries further support the synthesis of EFN in the secretory cells of the nectary. Chapter 4: Nutritional benefits from domatia-inhabitants in an ant–plant interaction: interlopers do pay the rent (published in Functional Ecology) In this chapter, we explore how a myrmecophytic system could evolve in the absence of protection benefits from the partner ants. We investigate non-protective benefits, specifically trophic (nutrient) benefits, from the protective and non-protective ants and other invertebrates to the host plant, using stable isotope techniques. We selected three representative inhabitant species for our analysis, viz., the protective ant T. albipes, a non-protective ant Crematogaster dorhni, and the arboreal earthworm Perionyx pullus. We observed that earthworms contributed approximately 9% while protective or non-protective ants contributed approximately 17% of the nitrogen to the plant tissues nearest to the domatium. We also observed from 15N labelling experiments that that nutrients from the domatia are not restricted solely to the domatia-bearing branch but could travel to distant non-domatia bearing branches as well. This study demonstrated for the first time that non-protective ants and non-ant invertebrates that inhabit the domatia, and hitherto referred to as interlopers, could be in a trophic mutualism with the host plant, thereby proposing the possibility of trophic mutualism as a factor for the evolution and maintenance of the domatia trait in addition to or in the absence of protection mutualism. It is also possible that fitness benefits of bearing domatia, acquired via trophic mutualism, could later facilitate the establishment of a specialised ant–plant protection mutualism. Chapter 5: Structure and development of the caulinary domatia of Humboldtia brunonis In this chapter, we investigate the morphology of domatia at different ontogenetic stages in order to understand the mode of development of the domatia. Our observations show that the domatium of H. brunonis is formed spontaneously near the terminal end of a growing branch, next to the young apical shoot. It appears as a young swollen internode which is soft and fleshy with the pith tissue still present. As the domatia grows and expands, the collective effect of both schizogeny as well as lysogeny, act on on the pith region. We also observed acropetal lignification of the pith cells around the hollow chamber. We investigate micro-scale anatomy of the inner wall of the domatia using scanning electron microscopy, and observed that the inner lining of the domatia cavity have canaliculated, lignified sclerenchyma with numerous plasmodesmata (intercellular pits) that could facilitate the flow of occupant-derived nutrients supporting trophic interaction between the plant and its domatia inhabitants. We also observed fungal mycelia-like structures in ant-occupied domatia that suggests the possibility of a fungus as a third party in the ant–plant trophic mutualism, as is observed in some other myrmecophytic systems. This aspect however needs further investigation. Chapter 6: Conclusion In this chapter, the main findings of the preceding chapters are summarised. A general conclusion of the thesis is provided, and future directions leading from the present thesis are also listed. The present thesis has explored the dynamics of interactions between a unique semi-myrmecophyte and its domatia-inhabitants; while the unprotected host plant resorts to an escape strategy to evade foliar herbivory, the ants (and other invertebrates) seem to have bid “a farewell to arms” and yet maintain a mutualism with its host via nutrient exchange. The results of this thesis contribute to furthering our current understanding of the evolution and stability of inter-species mutualisms.