Obligate Pollination Mutualism Research Articles

Absence of population‐level phenotype matching in an obligate pollination mutualism

Coevolution is thought to promote evolutionary change between demes that ultimately results in speciation. If this is the case, then we should expect to see similar patterns of trait matching and phenotypic divergence between populations and between species in model systems for coevolution. As measures of divergence are frequently only available at one scale (population level or taxon level), this contention is rarely tested directly. Here, we use the case of co-divergence between different varieties of Joshua tree Yucca brevifolia (Agavaceae) and their obligate pollinators, two yucca moths (Tegeticula spp. Prodoxidae), to test for trait matching between taxa and among populations. Using model selection, we show that there is trait matching between mutualists at the taxon level, but once we account for differences between taxa, there is no indication of trait matching in local populations. This result differs from similar studies in other coevolving systems. We hypothesize that this discrepancy arises because coevolution in obligate mutualisms favours divergence less strongly than coevolution in other systems, such as host–parasite interactions.

How to become a yucca moth: minimal trait evolution needed to establish the obligate pollination mutualism

The origins of obligate pollination mutualisms, such as the classic yucca-yucca moth association, appear to require extensive trait evolution and specialization. To understand the extent to which traits truly evolved as part of establishing the mutualistic relationship, rather than being preadaptations, we used an expanded phylogenetic estimate with improved sampling of deeply-diverged groups to perform the first formal reconstruction of trait evolution in pollinating yucca moths and their non-pollinating relatives. Our analysis demonstrates that key life history traits of yucca moths, including larval feeding in the floral ovary and the associated specialized cutting ovipositor, as well as colonization of woody monocots in xeric habitats, may have been established before the obligate mutualism with yuccas. Given these preexisting traits, novel traits in the mutualist moths are limited to the active pollination behaviors and the tentacular appendages that facilitate pollen collection and deposition. These results suggest that a highly specialized obligate mutualism was built on the foundation of preexisting interactions between early Prodoxidae and their host plants, and arose with minimal trait evolution.

Mutualism favours higher host specificity than does antagonism in plant–herbivore interaction

Coevolved mutualisms often exhibit high levels of partner specificity. Obligate pollination mutualisms, such as the fig-fig wasp and yucca-yucca moth systems, represent remarkable examples of such highly species-specific associations; however, the evolutionary processes underlying these patterns are poorly understood. The prevailing hypothesis suggests that the high degree of specificity in pollinating seed parasites is the fortuitous result of specialization in their ancestors because these insects are derived from endophytic herbivores that are themselves highly host-specific. Conversely, we show that in the Glochidion-Epicephala obligate pollination mutualism, pollinators are more host-specific than are closely related endophytic leaf-feeding taxa, which co-occur with Epicephala on the same Glochidion hosts. This difference is probably not because of shifts in larval diet (i.e. from leaf- to seed-feeding), because seed-eating lepidopterans other than Epicephala do not show the same degree of host specificity as Epicephala. Species of a tentative sister group of Epicephala each attack several distantly related plants, suggesting that the evolution of strict host specificity is tied to the evolution of pollinator habit. These results suggest that mutualists can attain higher host specificity than that of their parasitic ancestors and that coevolutionary selection can be a strong promoter of extreme reciprocal specialization in mutualisms.

Evolution of obligate pollination mutualism in the tribe Phyllantheae (Phyllanthaceae)

AbstractThe landmark discovery of obligate pollination mutualism between Glochidion plants and Epicephala moths has sparked increased interest in the pollination systems of Phyllantheae plants. In this paper I review current information on the natural history and evolutionary history of obligate pollination mutualism in Phyllantheae. Currently, an estimated >500 species are mutualistic with Epicephala moths that actively pollinate flowers and whose progeny feed on the resulting seeds. The Phyllantheae also includes species that are not mutualistic with Epicephala moths and are instead pollinated by bees and/or flies or ants. Phylogenetic analyses indicate that the mutualism evolved independently five times within Phyllantheae, whereas active pollination behavior, a key innovation in this mutualism, evolved once in Epicephala. Reversal of mutualism has occurred at least once in both partner lineages, involving a Breynia species that evolved an alternative pollination system and a derived clade of Epicephala that colonized ant‐pollinated Phyllantheae hosts and thereby lost the pollinating habit. The plant–moth association is highly species specific, although a strict one‐to‐one assumption is not perfectly met. A comparison of plant and moth phylogenies suggests signs of parallel speciation, but partner switches have occurred repeatedly at a range of taxonomic levels. Overall, the remarkable species diversity and multiple originations of the mutualism provide excellent opportunities to address many important questions on mutualism and the coevolutionary process. Although research on the biology of the mutualism is still in its infancy, the Phyllantheae–Epicephala association holds promise as a new model system in ecology and evolutionary biology.

Host specificity and reproductive success of yucca moths (Tegeticula spp. Lepidoptera: Prodoxidae) mirror patterns of gene flow between host plant varieties of the Joshua tree (Yucca brevifolia: Agavaceae)

Coevolution between flowering plants and their pollinators is thought to have generated much of the diversity of life on Earth, but the population processes that may have produced these macroevolutionary patterns remain unclear. Mathematical models of coevolution in obligate pollination mutualisms suggest that phenotype matching between plants and their pollinators can generate reproductive isolation. Here, we test this hypothesis using a natural experiment that examines the role of natural selection on phenotype matching between yuccas and yucca moths (Tegeticula spp.) in mediating reproductive isolation between two varieties of Joshua tree (Yucca brevifolia var. brevifolia and Y. brevifolia var. jaegeriana). Using passive monitoring techniques, DNA barcoding, microsatellite DNA genotyping, and sibship reconstruction, we track host specificity and the fitness consequences of host choice in a zone of sympatry. We show that the two moth species differ in their degree of host specificity and that oviposition on a foreign host plant results in the production of fewer offspring. This difference in host specificity between the two moth species mirrors patterns of chloroplast introgression from west to east between host varieties, suggesting that natural selection acting on pollinator phenotypes mediates gene flow and reproductive isolation between Joshua-tree varieties.

Private channel: a single unusual compound assures specific pollinator attraction in Ficus semicordata

Summary 1. Floral scents have been suggested to play a key role in the obligate pollination mutualism between figs and fig wasps. However, few studies have determined whether pollinator‐attractive compounds could alone assure species‐specificity (‘private channel’), or whether specificity is mediated by more complex ‘floral filters’, of which scent is only one component. 2. We examined changes in the floral volatile compounds of Ficus semicordata, a dioecious fig species, during and after pollination using headspace collection and compound identification by Gas Chromatography/Mass Spectrometry (GC/MS). One benzenoid compound, 4‐methylanisole, was strongly predominant (94–98%) among the volatile compounds emitted by both male and female receptive figs of F. semicordata, whereas it was totally absent in the volatiles emitted by figs 4 days after pollination, as well as in receptive‐stage volatiles emitted by two other sympatric fig species, Ficus racemosa and Ficus hispida. 3. Bioassays using the specific pollinator of F. semicordata, Ceratosolen gravelyi, in a Y‐tube olfactometer showed that 4‐methylanisole was attractive to C. gravelyi in a wide range of concentrations (from 1·22 × 10−2 ng/100 μL to 1·22 × 106ng/100μL). Moreover, chemical blends lacking 4‐methylanisole were unattractive to C. gravelyi. These non‐active odour sources included volatile compounds emitted by receptive figs of the two other sympatric fig species and volatiles of F. semicordata post‐pollination figs. 4. All these results suggest that 4‐methylanisole is the main signal compound in the floral scent of F. semicordata that attracts its obligate pollinator to the host figs at the precise stage required for pollination and oviposition. Furthermore, the high proportion of 4‐methylanisole in the odours of receptive figs of both sexes was consistent with the hypothesis of chemical mimicry in dioecious figs. 5. A simple signal comprised of one compound that is unusual among Ficus and that is an infrequent, usually minor, component of other floral odours, may thus function as a private channel in this specialized obligate mutualism.

Divergence in an obligate mutualism is not explained by divergent climatic factors

Adaptation to divergent environments creates and maintains biological diversity, but we know little about the importance of different agents of ecological divergence. Coevolution in obligate mutualisms has been hypothesized to drive divergence, but this contention has rarely been tested against alternative ecological explanations. Here, we use a well-established example of coevolution in an obligate pollination mutualism, Yucca brevifolia and its two pollinating yucca moths, to test the hypothesis that divergence in this system is the result of mutualists adapting to different abiotic environments as opposed to coevolution between mutualists. We used a combination of principal component analyses and ecological niche modeling to determine whether varieties of Y. brevifolia associated with different pollinators specialize on different environments. Yucca brevifolia occupies a diverse range of climates. When the two varieties can disperse to similar environments, they occupy similar habitats. This suggests that the two varieties have not specialized on distinct habitats. In turn, this suggests that nonclimatic factors, such as the biotic interaction between Y. brevifolia and its pollinators, are responsible for evolutionary divergence in this system.

Repeated independent evolution of obligate pollination mutualism in the Phyllantheae– Epicephala association

The well-known fig-fig wasp and yucca-yucca moth mutualisms are classic examples of obligate mutualisms that have been shaped by millions of years of coevolution. Pollination systems involving obligate seed parasites are only expected to evolve under rare circumstances where their positive effects are not swamped by abundant co-pollinators and heavy costs resulting from seed destruction. Here, we show that, in Phyllantheae, specialization to pollination by Epicephala moths evolved at least five times, involving more than 500 Phyllantheae species in this obligate association. Active pollination behaviour evolved once in Epicephala, 10-20 Myr after the initial divergence of their host plants. The pollinating Epicephala moths thus radiated on an already-diverged host lineage and successively colonized new Phyllantheae hosts, thereby giving rise to repeated independent evolution of the specialized pollination system in Phyllantheae. The present evolutionary success of this association rests entirely upon active pollination by Epicephala, making this a distinct example of an evolutionary key innovation. Overall, our findings provide a clear empirical demonstration of how a combination of evolutionary innovation and partner shifts facilitates the spread of mutualism in a coevolving species interaction.

DISTINGUISHING COEVOLUTION FROM COVICARIANCE IN AN OBLIGATE POLLINATION MUTUALISM: ASYNCHRONOUS DIVERGENCE IN JOSHUA TREE AND ITS POLLINATORS

Obligate pollination mutualisms--in which both plants and their pollinators are reliant upon one another for reproduction--represent some of the most remarkable coevolutionary interactions in the natural world. The intimacy and specificity of these interactions have led to the prediction that the plants and their pollinators may be prone to cospeciation driven by coevolution. Several studies have identified patterns of phylogenetic congruence that are consistent with this prediction, but it is difficult to determine the evolutionary process that underlies these patterns. Phylogenetic congruence might also be produced by extrinsic factors, such as a shared biogeographic history. We examine the biogeographic history of a putative case of codivergence in the obligate pollination mutualism between Joshua trees (Yucca brevifolia) and two sister species of pollinating yucca moths (Tegeticula spp.) We employ molecular phylogenetic methods and coalescent-based approaches, in combination with relaxed-clock estimates of absolute rates of molecular evolution, to analyze multi-locus sequence data from more than 30 populations of Y. brevifolia and its pollinators. The results indicate that the moth species diverged significantly (p < 0.01) more recently than their corresponding host populations, suggesting that the apparent codivergence is not an artifact of a shared biogeographic history.

Coevolution and Divergence in the Joshua Tree/Yucca Moth Mutualism

Theory suggests that coevolution drives diversification in obligate pollination mutualism, but it has been difficult to disentangle the effects of coevolution from other factors. We test the hypothesis that differential selection by two sister species of pollinating yucca moths (Tegeticula spp.) drove divergence between two varieties of the Joshua tree (Yucca brevifolia) by comparing measures of differentiation in floral and vegetative features. We show that floral features associated with pollination evolved more rapidly than vegetative features extrinsic to the interaction and that a key floral feature involved in the mutualism is more differentiated than any other and matches equivalent differences in the morphology of the pollinating moths. A phylogenetically based, ancestral states reconstruction shows that differences in moth morphology arose in the time since they first became associated with Joshua trees. These results suggest that coevolution, rather than extrinsic environmental factors, has driven divergence in this obligate pollination mutualism.

Pattern and timing of diversification in Yucca (Agavaceae): specialized pollination does not escalate rates of diversification

The yucca-yucca moth interaction is one of the most well-known and remarkable obligate pollination mutualisms, and is an important study system for understanding coevolution. Previous research suggests that specialist pollinators can promote rapid diversification in plants, and theoretical work has predicted that obligate pollination mutualism promotes cospeciation between plants and their pollinators, resulting in contemporaneous, parallel diversification. However, a lack of information about the age of Yucca has impeded efforts to test these hypotheses. We used analyses of 4322 AFLP markers and cpDNA sequence data representing six non-protein-coding regions (trnT-trnL, trnL, trnL intron, trnL-trnF, rps16 and clpP intron 2) from all 34 species to recover a consensus organismal phylogeny, and used penalized likelihood to estimate divergence times and speciation rates in Yucca. The results indicate that the pollination mutualism did not accelerate diversification, as Yucca diversity (34 species) is not significantly greater than that of its non-moth-pollinated sister group, Agave sensu latissimus (240 species). The new phylogenetic estimates also corroborate the suggestion that the plant-moth pollination mutualism has at least two origins within the Agavaceae. Finally, age estimates show significant discord between the age of Yucca (ca 6-10Myr) and the current best estimates for the age of their pollinators (32-40Myr).

Odor-Mediated Push-Pull Pollination in Cycads

The reproductive organs of some plants self-heat, release scent, and attract pollinators. The relations among these processes are not well understood, especially in the more ancient, nonflowering gymnosperm lineages. We describe the influence of plant volatiles in an obligate pollination mutualism between an Australian Macrozamia cycad (a gymnosperm with male and female individuals) and its specialist thrips pollinator, Cycadothrips chadwicki. Pollen-laden thrips leave male cycad cones en masse during the daily thermogenic phase, when cone temperatures and volatile emissions increase dramatically and thrips are repelled. As thermogenesis declines, total volatile emissions diminish and cones attract thrips, resulting in pollination of female cones. Behavioral and electrophysiological tests on thrips reveal that variations in b-myrcene and ocimene emissions by male and female cones are sufficient to explain the observed sequential thrips' repellence (push) and attraction (pull). These dynamic interactions represent complex adaptations that enhance the likelihood of pollination and may reflect an intermediate state in the evolution of biotic pollination.

Strong Conservation of Floral Scent Composition in Two Allopatric Yuccas

Floral scent has been suggested to play a key role in the obligate pollination mutualism between yuccas and yucca moths. We analyzed floral fragrance compounds of Yucca elata with headspace collection followed by gas chromatography and mass spectrometry, and compared the odor blend with the recently characterized blend of the allopatric Yucca filamentosa. A principal component analysis based on 20 scent compounds revealed that the floral odor bouquets of Y. elata and Y. filamentosa are virtually identical. Although the two plants belong to the same section of capsular-fruited yuccas, they rely on different species of Tegeticula moths for pollination and probably have been allopatric for several million years. Yet, their floral odor blends are very similar, which may indicate that strong selection by obligate pollinators counteracts drift or divergence in this trait.

Impact of the Third Trophic Level in an Obligate Mutualism: Do Yucca Plants Benefit from Parasitoids of Yucca Moths?

In obligate pollination mutualism, selection should favor increased exploitation by either party at the expense of the other. One potential pathway to increase such exploitation at the expense of the mutualist partner is through indirect mutualism across trophic levels, where the natural enemy of a pollinator could reduce pollination costs to the plant. We tested this hypothesis, asking whether a specialized parasitoid wasp that kills yucca moth larvae before they have completed seed feeding will increase the production of intact yucca seeds. In the two years of study in a population of Yucca treculeana Carriere (Agavaceae) in west Texas, per capita seed consumption by parasitized larvae of both a pollinator and a nonpollinator yucca moth were significantly reduced when compared to that of nonparasitized conspecific larvae. In one year, fruits that contained at least some parasitized larvae produced significantly more intact seeds than ones with only nonparasitized larvae, whereas the opposite effect was ...

COMMUNITY CONTEXT OF AN OBLIGATE MUTUALISM: POLLINATOR AND FLORIVORE EFFECTS ON YUCCA FILAMENTOSA

Obligate pollination mutualisms have been central to our understanding of the ecology and evolution of mutualisms. Although usually viewed as pairwise interactions, obligate mutualists also interact with other community members that may impact the mu- tualism. In this study, we examined the community context of the obligate mutualism between the plant Yucca filamentosaand its yucca moth pollinator Tegeticula cassandra to determine the relationships among plant traits, the community assemblage, and fruit set. In addition to attracting its pollinator, Y. filamentosain central Florida also attracts two insect florivores: the hemipteran Leptoglossus phyllopusand the beetle Hymenorus densus. We surveyed flowering Y. filamentosain 2001 and 2002 to determine the abundance of the three insect species throughout the flowering period. We also measured fruit set and four plant traits thought to be important in attracting the moth pollinator. Path analysis revealed that T. cassandra was the most important determinant of relative fruit set for both years. Peak pollinator abundance was significantly earlier than peak flowering. Consequently, we found a negative relationship between plant flowering date and pollinator abundance. Fur- thermore, taller plants also attracted more pollinators. Leptoglossus phyllopus had a sig- nificantly negative impact on relative fruit set in 2001 but not in 2002, whereas H. densus had no effect in either year. Abundance of the two florivores was more closely tied to peak flowering of Y. filamentosathan was the peak abundance of the pollinator moths. Although both florivore species reached high densities on Y. filamentosa , their effect on relative fruit set through floral abscission may be partially masked by the characteristically high ab- scission rates found in yuccas. As a result, florivores may be feeding on flowers that are already destined to be abscised. Similarly, pollinator moths may be preadapted to handle the effects of florivores because females lay eggs across multiple plants and flowers. Hence, the obligate mutualism between yuccas and yucca moths may be somewhat buffered from the effects of local community members depending on whether florivores substantially increase floral abscission rates from year to year.

Phylogeny and life history evolution of Prodoxus yucca moths (Lepidoptera: Prodoxidae)

Abstract. Yucca moths (Lep., Prodoxidae) are well‐known for their obligate pollination mutualism with yuccas. In addition to the pollinators, yuccas also host many non‐pollinating yucca moths. Here the genus Prodoxus, the non‐pollinating sister group of the pollinators, is revised using morphological and molecular data, their phylogenetic relationships are analysed, and the evolution of host tissue specialization explored. Twenty‐two species are recognized, including nine new species: Prodoxus gypsicolorsp.n., P. sonorensissp.n., P. carnerosanellussp.n., P. tamaulipellussp.n., P. weethumpisp.n., P. tehuacanensissp.n., P. californicussp.n., P. mapimiensissp.n. and P. atascosanellussp.n.Prodoxus y‐inversus Riley, P. coloradensis Riley and P. sordidus Riley are redescribed. The genus Agavenema is synonymized with Prodoxus. Phylogenetic analyses indicated that stalk‐feeding is basal within the group, that there are three separate origins of fruit‐feeding, and one origin of leaf‐mining from a stalk‐feeding ancestor. Although species with different feeding habits often coexist within hosts, the analyses suggest that ecological specialization and diversification within a host only may have occurred within one or possibly two hosts.

TESTING THE OUT-OF-FLORIDA HYPOTHESIS ON THE ORIGIN OF CHEATING IN THE YUCCA YUCCA MOTH MUTUALISM

Mutualistic interactions can be exploited by cheaters that take the rewards offered by mutualists without providing services in return. The evolution of cheater species from mutualist ancestors is thought to be possible under particular ecological conditions. Here we provide a test of the first explicit model of the transition from mutualism to antagonism. We used the obligate pollination mutualism between yuccas and yucca moths to examine the origins of a nonpollinating cheater moth, Tegeticula intermedia, and its pollinating sister species, T. cassandra. Based on geographic distribution and ecological factors affecting the pollinators, previous research had indicated that the cheaters evolved in Florida as a result of sympatry of T. cassandra and another pollinator species. We used mitochondrial DNA (mtDNA) sequences and amplified fragment length polymorphism (AFLP) data to investigate the phylogeographic history of the pollinator-cheater sister pair and to test whether the cheaters arose in Florida. Contrary to predictions, phylogenetic and population genetic analyses suggested that the cheaters evolved in the western United States and subsequently spread eastward. Western populations of cheaters had the most ancestral haplotypes and the highest genetic diversity, and there was also significant genetic structure associated with a geographic split between eastern and western populations. In comparison, there was evidence for weak genetic structure between northern and southern pollinator populations, suggesting a long history in Florida. The western origin of the cheaters indicated that the pollinators have more recently become restricted to the southeastern United States. This was supported by AFLP analyses that indicated that the pollinators were more closely related to the western cheaters than they were to geographically proximate cheaters in the east. Shared mtDNA between pollinators and eastern cheaters suggested hybridization, possibly in a secondary contact zone. The results negate the out-of-Florida hypothesis and reveal instead a long, complex, and disparate history for the pollinator-cheater sister pair.

COSPECIATION ANALYSIS OF AN OBLIGATE POLLINATION MUTUALISM: HAVEGLOCHIDION TREES (EUPHORBIACEAE) AND POLLINATING EPICEPHALA MOTHS(GRACILLARIIDAE) DIVERSIFIED IN PARALLEL?

Species-specific obligate pollination mutualism between Glochidion trees (Euphorbiaceae) and Epicephala moths (Gracillariidae) involves a large number of interacting species and resembles the classically known fig-fig wasp and yucca-yucca moth associations. To assess the extent of parallel cladogenesis in Glochidion-Epicephala association, we reconstruct phylogenetic relationships of 18 species of Glochidion using nuclear ribosomal DNA sequences (internal and external transcribed spacers) and those of the corresponding 18 Epicephala species using mitochondrial (the cytochrome oxidase subunit I gene) and nuclear DNA sequences (the arginine kinase and elongation factor-1alpha genes). Based on the obtained phylogenies, we determine whether Glochidion and Epicephala have undergone parallel diversification using several different methods for investigating the level of cospeciation between phylogenies. These tests indicate that there is generally a greater degree of correlation between Glochidion and Epicephala phylogenies than expected in a random association, but the results are sensitive to selection of different phylogenetic hypotheses and analytical methods for evaluating cospeciation. Perfect congruence between phylogenies is not found in this association, which likely resulted from host shift by the moths. The observed significant discrepancy between Glochidion and Epicephala phylogenies implies that the one-to-one specificity between the plants and moths has been maintained through a complex speciation process or that there is an underestimated diversity of association between Glochidion trees and Epicephala moths.

Obligate pollination mutualism in Breynia (Phyllanthaceae): further documentation of pollination mutualism involving Epicephala moths (Gracillariidae)

This paper reports obligate seed-parasitic pollination mutualisms in Breynia vitis-idea and B. fruticosa (Phyllanthaceae). The genus Breynia is closely related to Glochidion and Gomphidium (a subgenus of Phyllanthus), in which pollination by species-specific, seed-parasitic Epicephala moths (Gracillariidae) have been previously reported. At night, female Epicephala moths carrying numerous pollen grains on their proboscises visited female flowers of B. vitis-idea, actively pollinated flowers, and each subsequently laid an egg. Examination of field-collected flowers indicated that pollinated flowers of B. vitis-idea and B. fruticosa almost invariably had Epicephala eggs, suggesting that these moths are the primary pollinators of the two species. Single Epicephala larvae consumed a fraction of seeds within developing fruit in B. vitis-idea and all seeds in B. fruticosa. However, some of the fruits were left untouched, and many of these had indication of moth oviposition, suggesting that egg/larval mortality of Epicephala moths is an important factor assuring seed set in these plants. The overall similarity of the specialized floral structure among Breynia species may indicate that this pollination system is fairly widespread within the genus.

Evolution of obligate pollination mutualism in New Caledonian Phyllanthus (Euphorbiaceae)

About half a dozen obligate pollination mutualisms between plants and their seed-consuming pollinators are currently recognized, including fig-fig wasp, yucca-yucca moth, and the recently discovered Glochidion tree-Epicephala moth mutualisms. A common principle among these interactions is that the pollinators consume only a limited amount of the seed crop within a developing fruit (or fig in the case of fig-fig wasp mutualism), thereby ensuring a net benefit to plant reproduction. A novel obligate, seed-parasitic pollination mutualism between two species of New Caledonian Phyllanthus (Euphorbiaceae), a close relative of Glochidion, and Epicephala moths (Gracillariidae) is an exception to this principle. The highly specialized flowers of Phyllanthus are actively and exclusively pollinated by species-specific Epicephala moths, whose larvae consume all six ovules of the developing fruit. Some flowers pollinated by the moths remain untouched, and thus a fraction of the fruits is left intact. Additional evidence for a similar association of Epicephala moths in other Phyllanthus species suggests that this interaction is a coevolved, species-specific pollination mutualism. Implications for the evolutionary stability of the system, as well as differences in mode of interaction with respect to the Glochidion-Epicephala mutualism, are discussed.