Evolution of microbial markets

Biological market theory has been used successfully to explain cooperative behavior in many animal species. Microbes also engage in cooperative behaviors, both with hosts and other microbes, that can be described in economic terms. However, a market approach is not traditionally used to analyze these interactions. Here, we extend the biological market framework to ask whether this theory is of use to evolutionary biologists studying microbes. We consider six economic strategies used by microbes to optimize their success in markets. We argue that an economic market framework is a useful tool to generate specific and interesting predictions about microbial interactions, including the evolution of partner discrimination, hoarding strategies, specialized versus diversified mutualistic services, and the role of spatial structures, such as flocks and consortia. There is untapped potential for studying the evolutionary dynamics of microbial systems. Market theory can help structure this potential by characterizing strategic investment of microbes across a diversity of conditions.

Related papers:

title: Biological markets: supply and demand determine the effect of partner choice in cooperation, mutualism and mating
abstract: The formation of collaborating pairs by individuals belonging to two different classes occurs in the contexts of reproduction and intea-specific cooperation as well as of inter-specific mutualism. There is potential for partner choice and for competition for access to preferred partners in all three contexts. These selective forces have long been recognised as important in sexual selection, but their impact is not yet appreciated in cooperative and mutualistic systems. The formation of partnerships between members of different classes has much in common with the conclusion of trade agreements in human markets with two classes of traders, like producers and consumers, or employers and employees. Similar game-theoretical models can be used to predict the behaviour of rational traders in human markets and the evolutionarily stable strategies used in biological markets. We present a formal model in which the influence of the market mechanism on selection is made explicit. We restrict ourselves to biological markets in which: (1) Individuals do not compete over access to partners in an agonistic manner, but rather by outcompeting each other in those aspects that are preferred by the choosing party. (2) The commodity the partner has to offer cannot be obtained by the use of force, but requires the consent of the partner. These two restrictions ensure a dominant role for partner choice in the formation of partnerships. In a biological market model the decision to cooperate is based on the comparison between the offers of several potential partners, rather than on the behaviour of a single potential partner, as is implicitly assumed in currently accepted models of cooperation. In our example the members of one class A offer a commodity of fixed value in exchange for a commodity of variable value supplied by the other class, B. We show that when the B-class outnumbers the A-class sufficiently and the cost for the A-class to sample the offers of the B-class are low, the choosiness of the A-class will lead to selection for the supply of high value commodities by the B-class . Under the same market conditions, but with a high sampling cost this may still be the evolutionariy stable outcome, but another pair of strategies proves to be stable too: relaxed choosiness of class A coupled with low value commodities supplied by class B (Fig. 3b). We give a number of examples of mating, cooperative and mutualistic markets that resemble the low sampling cost situation depicted in Fig. 3a.

title: Biological markets.
abstract: In biological markets, two classes of traders exchange commodities to their mutual benefit. Characteristics of markets are: competition within trader classes by contest or outbidding; preference for partners offering the highest value; and conflicts over the exchange value of commodities. Biological markets are currently studied under at least three different headings: sexual selection, intraspecific cooperation and interspecific mutualism. The time is ripe for the development of game theoretic models that describe the common core of biological markets and integrate existing knowledge from the separate fields.

title: Selection for protection in an ant–plant mutualism: host sanctions, host modularity, and the principal–agent game
abstract: Retaliation against cheaters can prevent the breakdown of cooperation. Here we ask whether the ant–plant Cordia nodosa is able to apply retaliatory sanctions against its ant symbiont Allomerus octoarticulatus, which patrols new shoots to prevent herbivory. We test the hypothesis that the modular design of C. nodosa physiologically ties the growth of housing (stem swellings known as domatia) to the successful development of the attached leaves. We experimentally simulated herbivory by cutting leaves from patrolled shoots and found that the domatia on such ‘cheated’ shoots suffered higher mortality and lower growth than did controls, evidence for a host sanction. On the other hand, patrolling is costly to the ant, and experiment shows that non-patrollers run a low risk of being sanctioned because most leaves (and the attached domatia) escape heavy herbivory even when patrollers are absent. This suggests that cheaters might enjoy a higher fitness than do mutualists, despite sanctions, but we find that patrolling provides a net fecundity benefit when the colony and plant exceed a minimum size, which requires sustained ant investment in patrolling. These results map directly onto the principal–agent (P–A) game from economics, which we suggest can be used as a framework for studying stability in mutualisms, where high sampling costs and cheating do not allow market effects to select for mutual benefits.

title: Mutualism, market effects and partner control
abstract: Intraspecific cooperation and interspecific mutualism often feature a marked asymmetry in the scope for exploitation. Cooperation may nevertheless persist despite one‐sided opportunities for cheating, provided that the partner vulnerable to exploitation has sufficient control over the duration of interaction. The effectiveness of the threat of terminating an encounter, however, depends upon the ease with which both the potential victim and the potential exploiter can find replacement partners. Here, we extend a simple, game‐theoretical model of this form of partner control to incorporate variation in the relative abundance of potential victims and exploiters, which leads to variation in the time required for individuals of each type to find a new partner. We show that such market effects have a dramatic influence on the stable level of exploitation (and consequent duration of interaction). As the relative abundance of victims decreases, they become less tolerant to exploitation, terminating encounters earlier (for a given level of exploitation), whereas exploiters behave in a more cooperative manner. As a result, the stable duration of interaction actually increases, despite the decreasing tolerance of the victims. Below a critical level of relative victim abundance, the model suggests that the cost of finding a replacement partner becomes so great that it does not pay to exploit at all.

title: Our Current Understanding of Mutualism
abstract: It is widely believed that mutualisms, interspecific interactions that benefit both species, have been grossly neglected relative to their true importance in nature. I have reviewed the recent primary literature in order to assess quentitatively the frequency of studies of mutualism, the types of questions they address, and their general scientific approach. All articles appearing from 1986 to 1990 in nine major journals that publish ecological and evolutionary research were examined. It is clear that mutualism research is not in fact rare. Studies of interspecific interactions made up about 22% of theover 4500 articles published during this period; of these, about one-quarter investigated some form of mutualism. Over 90% of them investigated plant-animal interactions, primarily pollination (52%) and seed dispersal (31%), a bias probably related in part to the particular journals examined. The diversity of questions addressed in these articles was surprisingly low. The majority (63%) focused simply on identifying the mutualist of some species of interest. Furthermore, almost all studies were unilateral, that is, they focused on only one of the interacting species, plants being studied much more frequently than their animal partners. Mutualism studies do not appear to have focused on mutualism as a form of interaction in the same way as studies of competition and predation. Rather, researchers have treated mutualism primarily as a life history attribute of one of the two partners. Consequently, although an impressive amount of information has accumulated about these interactions, we are still far from achieving an overall picture that transcends the boundaries of particular taxa or comibinations of taxa. Three other obstacles have prevented data on mutualisms from being brought together: the historical isolation of studies of different kinds of mutualism, a nearly total disconnection between mutualism theories and empirical studies, and the unilateral approach almost always used to study these bilateral interactions. I identify eight research questions whose answers have the potential to reveal broad-based generalizations about the evolution and ecology of mutualism.

title: Animals in a bacterial world, a new imperative for the life sciences
abstract: In the last two decades, the widespread application of genetic and genomic approaches has revealed a bacterial world astonishing in its ubiquity and diversity. This review examines how a growing knowledge of the vast range of animal–bacterial interactions, whether in shared ecosystems or intimate symbioses, is fundamentally altering our understanding of animal biology. Specifically, we highlight recent technological and intellectual advances that have changed our thinking about five questions: how have bacteria facilitated the origin and evolution of animals; how do animals and bacteria affect each other’s genomes; how does normal animal development depend on bacterial partners; how is homeostasis maintained between animals and their symbionts; and how can ecological approaches deepen our understanding of the multiple levels of animal–bacterial interaction. As answers to these fundamental questions emerge, all biologists will be challenged to broaden their appreciation of these interactions and to include investigations of the relationships between and among bacteria and their animal partners as we seek a better understanding of the natural world.

title: The rhizosphere microbiome and plant health.
abstract: The diversity of microbes associated with plant roots is enormous, in the order of tens of thousands of species. This complex plant-associated microbial community, also referred to as the second genome of the plant, is crucial for plant health. Recent advances in plant-microbe interactions research revealed that plants are able to shape their rhizosphere microbiome, as evidenced by the fact that different plant species host specific microbial communities when grown on the same soil. In this review, we discuss evidence that upon pathogen or insect attack, plants are able to recruit protective microorganisms, and enhance microbial activity to suppress pathogens in the rhizosphere. A comprehensive understanding of the mechanisms that govern selection and activity of microbial communities by plant roots will provide new opportunities to increase crop production.

title: Root nodulation: a paradigm for how plant-microbe symbiosis influences host developmental pathways.
abstract: Legume plants have an exceptional capacity for association with microorganisms, ranging from largely nonspecific to very specific interactions. Legume-rhizobial symbiosis results in major developmental and metabolic changes for both the microorganism and host, while providing the plant with fixed nitrogen. A complex signal exchange leads to the selective rhizobial colonization of plant cells within nodules, new organs that develop on the roots of host plants. Although the nodulation mechanism is highly specific, it involves the same subset of plant phytohormones, namely auxin, cytokinin, and ethylene, which are required for root development. In addition, nodulation triggered by the rhizobia affects the development of the host root system, indicating that the microorganism can alter host developmental pathways. Nodulation by rhizobia is a prime example of how microorganisms and plants have coevolved and exemplifies how microbial colonization may affect plant developmental pathways.

title: The Social Lives of Microbes
abstract: Our understanding of the social lives of microbes has been revolutionized over the past 20 years. It used to be assumed that bacteria and other microorganisms lived relatively independent unicellular lives, without the cooperative behaviors that have provoked so much interest in mammals, birds, and insects. However, a rapidly expanding body of research has completely overturned this idea, showing that microbes indulge in a variety of social behaviors involving complex systems of cooperation, communication, and synchronization. Work in this area has already provided some elegant experimental tests of social evolutionary theory, demonstrating the importance of factors such as relatedness, kin discrimination, competition between relatives, and enforcement of cooperation. Our aim here is to review these social behaviors, emphasizing the unique opportunities they offer for testing existing evolutionary theory as well as highlighting the novel theoretical problems that they pose.

title: Monkeys benefit from reciprocity without the cognitive burden
abstract: The debate about the origins of human prosociality has focused on the presence or absence of similar tendencies in other species, and, recently, attention has turned to the underlying mechanisms. We investigated whether direct reciprocity could promote prosocial behavior in brown capuchin monkeys (Cebus apella). Twelve capuchins tested in pairs could choose between two tokens, with one being “prosocial” in that it rewarded both individuals (i.e., 1/1), and the other being “selfish” in that it rewarded the chooser only (i.e., 1/0). Each monkey’s choices with a familiar partner from their own group was compared with choices when paired with a partner from a different group. Capuchins were spontaneously prosocial, selecting the prosocial option at the same rate regardless of whether they were paired with an in-group or out-group partner. This indicates that interaction outside of the experimental setting played no role. When the paradigm was changed, such that both partners alternated making choices, prosocial preference significantly increased, leading to mutualistic payoffs. As no contingency could be detected between an individual’s choice and their partner’s previous choice, and choices occurred in rapid succession, reciprocity seemed of a relatively vague nature akin to mutualism. Having the partner receive a better reward than the chooser (i.e., 1/2) during the alternating condition increased the payoffs of mutual prosociality, and prosocial choice increased accordingly. The outcome of several controls made it hard to explain these results on the basis of reward distribution or learned preferences, and rather suggested that joint action promotes prosociality, resulting in so-called attitudinal reciprocity.

title: Extent and limits of cooperation in animals
abstract: Individuals in many animal species are strongly motivated to form close social bonds and to attend to the social interactions of others. Some animals may also recognize other individuals’ intentions and simple mental states. Such curiosity appears to be adaptive, because it enables observers to learn about others’ status and relationships and to anticipate future events without direct participation. However, many questions remain unresolved. In particular, it remains unclear whether animals keep track of favors given and received when interacting with others, and whether they rely on memory of past cooperative acts when anticipating future ones. Primates appear to possess many of the cognitive abilities required for human-like contingent cooperation. However, most investigations of captive primates have indicated that cooperation is seldom contingency-based, and that interactions are not influenced by inequity aversion or sensitivity to cheaters. In contrast, several experiments with nonprimates have found that animals can take into account recent interactions when supporting others, suggesting that the apparent rarity of contingent cooperation in primates may not stem from cognitive constraints. Instead, individuals may tolerate short-term inequities in favors given and received because most cooperation occurs among long-term reciprocating partners.

title: Cooperation experiments: coordination through communication versus acting apart together
abstract: Over the last two decades a large number of experiments have been conducted in which two conspecifics, mainly vertebrates, could obtain rewards through cooperation. At least three different motivations to perform such experiments can be distinguished: (1) detecting the mechanistic basis of naturally occurring forms of cooperation; (2) analysing behavioural strategies specific to cooperation; and (3) testing game-theoretical models. Experimenters in the latter two categories, on which this review concentrates, make use of highly artificial devices. The impression of cooperation in the third category is created because two animals are tested simultaneously after having been trained to interact with an apparatus individually before the experiment. These results can be explained most parsimoniously as the product of instrumental (‘trial-and-error’) learning, hence my label ‘instrumental cooperation’. The underlying philosophy of several studies is reminiscent of the strongly disputed ideas of the ‘behaviourist school’. Results from experiments in the second category highlight the importance of communication; therefore I call this ‘communicative cooperation’. The crucial role of communication identified by studies in the second category calls into question the relevance of the most popular paradigm used in the third category, the two-player Iterated Prisoner's Dilemma. Its stringent conditions, which are hardly ever fulfilled in nature, are very difficult to fulfil in experiments too. I propose that future studies concentrate on experiments that emphasize the role of communication and partner choice with the aim of explaining the emergence and maintenance of naturally occurring forms of cooperation.

title: The interplay of cognition and cooperation
abstract: Cooperation often involves behaviours that reduce immediate payoffs for actors. Delayed benefits have often been argued to pose problems for the evolution of cooperation because learning such contingencies may be difficult as partners may cheat in return. Therefore, the ability to achieve stable cooperation has often been linked to a species' cognitive abilities, which is in turn linked to the evolution of increasingly complex central nervous systems. However, in their famous 1981 paper, Axelrod and Hamilton stated that in principle even bacteria could play a tit-for-tat strategy in an iterated Prisoner's Dilemma. While to our knowledge this has not been documented, interspecific mutualisms are present in bacteria, plants and fungi. Moreover, many species which have evolved large brains in complex social environments lack convincing evidence in favour of reciprocity. What conditions must be fulfilled so that organisms with little to no brainpower, including plants and single-celled organisms, can, on average, gain benefits from interactions with partner species? On the other hand, what conditions favour the evolution of large brains and flexible behaviour, which includes the use of misinformation and so on? These questions are critical, as they begin to address why cognitive complexity would emerge when ‘simple’ cooperation is clearly sufficient in some cases. This paper spans the literature from bacteria to humans in our search for the key variables that link cooperation and deception to cognition.

title: The economics of mutualisms: optimal utilization of mycorrhizal mutualistic partners by plants.
abstract: Can choice of mutualistic partners and the degree of their utilization determine (1) mutualistic partner coexistence, (2) relative abundance of mutualistic partners, and (3) environment-dependent changes in relative abundance? We investigate these questions in the context of the plant-mycorrhizal fungal mutualism by building a biological market model potentially applicable to other mutualisms as well. We examine the situation where a single plant selectively utilizes member(s) of a group of ectomycorrhizal potential trading partners. Under biologically realistic circumstances, the plant may simultaneously utilize multiple partners, its degree of utilization determining the community structure of the fungi. If utilization of multiple partners is optimal, the marginal cost of acquiring additional nitrogen from every trading partner must be equal while the marginal cost of acquiring it from any unutilized partner must be larger. Because the plant's nitrogen demand is light dependent, the composition of the fungal species among its trading partners changes along light-availability gradients. We discuss the design of an experiment to test the key prediction of our model, the equalization of marginal cost.

title: SPECIALIZATION AND RESOURCE TRADE: BIOLOGICAL MARKETS AS A MODEL OF MUTUALISMS
abstract: Most ecological theory suggests that the conditions that would give rise to the evolution of mutualisms are rare. In contrast, empirical evidence suggests that mutualisms are common. Thus, there appears to be a need for additional theory to describe conditions under which mutualisms may evolve. Furthermore, there is a need for theory to predict the conditions under which we expect interactions to remain mutualistic once established. We adopt a biological market approach to present a model for the evolution of resource exchange mutualisms, using the relationship between plants and mycorrhizal fungi as an example. We apply the economic theory of relative advantage to investigate the conditions under which species ought to specialize and trade. A simple economic analogy demonstrates that, in a two-resource model, a species that is relatively efficient at acquiring one resource would benefit from specialization on acquisition of that resource accompanied by trade for the other resource. The theory of relative advantage extends this prediction to show that specialization and trade confer an advantage even for species that are relatively poor resource competitors for both resources. Under the assumptions of our model, we show that two species ought to specialize in the acquisition of one resource and trade for a second resource as long as each species perceives different relative acquisition costs for the two resources. We also describe the conditions under which changing resource availabilities will benefit, or harm, both partners in a mutualism. We predict conditions conducive to mutualisms to occur when the costs of resource exchange are low, the opportunity to ensure fair trade is high, or the cost of tolerating cheaters is low. Market models such as ours may help to explain the conditionality often observed in mutualisms.

title: A multi-mutualist simulation: Applying biological market models to diverse mycorrhizal communities
abstract: We present a cellular automaton that simulates the interaction between a host tree and multiple potential mycorrhizal symbionts and generates testable hypotheses of how processes at the scale of individual root tips may explain mycorrhizal community composition. Existing theoretical biological market models imply that a single host is able to interact with and select from multiple symbionts to organize an optimal symbiont community. When evaluating the tree–symbiont interaction, two scales must be considered simultaneously: the scale of the entire host plant at which carbon utilization and nutrient demands operate, and the scale of the individual root tip, at which colonization and carbon-nutrient trade occurs. Three strategies that may be employed by the host tree for optimizing carbon use and nutrient acquisition through mycorrhizal symbiont communities are simulated: (1) carbon pool adjustment, in which the plant controls only the total amount of carbon to be distributed uniformly throughout the root system, (2) symbiont selection, wherein the plant opts either for or against the interaction at each fine root tip, and (3) selective carbon allocation, wherein the plant adjusts the amount of carbon allocated to each root tip based on the cost of nutrients. Strategies were tested over various nutrient availabilities (the amount of inorganically and organically bound nutrients). Success was defined on the basis of minimizing carbon expended for nutrient acquisition because this would allow more carbon to be utilized for growth and reproduction. In all cases, the symbiont selection and selective carbon allocation strategies were able to meet the nutritional requirements of the plant, but did not necessarily optimize carbon use. The carbon pool adjustment strategy is the only strategy that does not operate at the individual root tip scale, and the strategy was not successful when inorganic nutrients were scarce since there is no mechanism to exclude suboptimal symbionts. The combination of the symbiont selection strategy and the carbon pool adjustment resulted in optimal carbon use and nutrient acquisition under all environmental conditions but result in monospecific symbiont assemblages. On the other hand, the selective carbon allocation strategy is the only strategy that maintained successful, multi-symbiont communities. The simulations presented here thus imply clear hypotheses about the effect of nutrient availability on symbiont selection and mycorrhizal community richness and composition.

title: Ecological Specialization and Trade Affect the Outcome of Negotiations in Mutualism
abstract: By definition, mutualisms involve the exchange of goods or services between partners. It has been shown that mutualism can grade into parasitism, but even when exchange is mutually beneficial, a conflict of interest remains because each partner benefits from reaping more benefits at a lower cost. Metaphorically, the partners negotiate the conditions of trade, the outcome of which will determine the net benefit to each partner. Each partner can adjust its allocation to self-provisioning while negotiating the ratio at which benefits are exchanged. To understand how these two features of trade affect mutualisms, we used the example of the plant–arbuscular mycorrhizal mutualism and modeled uptake and trade of two resources, phosphorus and carbon. In most contexts, the fungus specialized on phosphorus uptake while the plant took up both phosphorus and carbon. However, when phosphorus was abundant and light was scarce, the plant specialized, taking up only carbon and relying on trade for phosphorus. Resource availability was the most important factor determining specialization and the outcome of negotiation and trade, but other aspects of the context were also important. These results suggest experiments to link these two key features of trade with environmental conditions to determine the outcome of mutualism.

title: Snowdrift game dynamics and facultative cheating in yeast
abstract: The origin of cooperation is a central challenge to our understanding of evolution. The fact that microbial interactions can be manipulated in ways that animal interactions cannot has led to a growing interest in microbial models of cooperation and competition. For the budding yeast Saccharomyces cerevisiae to grow on sucrose, the disaccharide must first be hydrolysed by the enzyme invertase. This hydrolysis reaction is performed outside the cytoplasm in the periplasmic space between the plasma membrane and the cell wall. Here we demonstrate that the vast majority (∼99 per cent) of the monosaccharides created by sucrose hydrolysis diffuse away before they can be imported into the cell, serving to make invertase production and secretion a cooperative behaviour. A mutant cheater strain that does not produce invertase is able to take advantage of and invade a population of wild-type cooperator cells. However, over a wide range of conditions, the wild-type cooperator can also invade a population of cheater cells. Therefore, we observe steady-state coexistence between the two strains in well-mixed culture resulting from the fact that rare strategies outperform common strategies—the defining features of what game theorists call the snowdrift game. A model of the cooperative interaction incorporating nonlinear benefits explains the origin of this coexistence. We are able to alter the outcome of the competition by varying either the cost of cooperation or the glucose concentration in the media. Finally, we note that glucose repression of invertase expression in wild-type cells produces a strategy that is optimal for the snowdrift game—wild-type cells cooperate only when competing against cheater cells.

title: Cooperation for direct fitness benefits
abstract: Studies of the evolution of helping have traditionally used the explanatory frameworks of reciprocity and altruism towards relatives, but recently there has been an increasing interest in other kinds of explanations. We review the success or otherwise of work investigating alternative processes and mechanisms, most of which fall under the heading of cooperation for direct benefits. We evaluate to what extent concepts such as by-product benefits, pseudo-reciprocity, sanctions and partner choice, markets and the build-up of cross-species spatial trait correlations have contributed to the study of the evolution of cooperation. We conclude that these alternative ideas are successful and show potential to further increase our understanding of cooperation. We also bring up the origin and role of common interest in the evolution of cooperation, including the appearance of organisms. We note that there are still unresolved questions about the main processes contributing to the evolution of common interest. Commenting on the broader significance of the recent developments, we argue that they represent a justified balancing of the importance given to different major hypotheses for the evolution of cooperation. This balancing is beneficial because it widens considerably the range of phenomena addressed and, crucially, encourages empirical testing of important theoretical alternatives.

title: Experimental evidence that partner choice is a driving force in the payoff distribution among cooperators or mutualists: the cleaner fish case
abstract: Supply and demand largely determine the price of goods on human markets. It has been proposed that in animals, similar forces influence the payoff distribution between trading partners in Sexual selection, intraspecific cooperation and interspecific mutualism. Here we present the first experimental evidence supporting biological market theory in it study on cleaner fish, Labroides dimidiatus. Cleaners interact with two classes of clients: choosy client species with access to several cleaners usually do not queue for service and do not return if ignored, while resident client species with access to only one cleaning station do queue or return. We used plexiglas plates with equal amounts of food to stimulate these behaviours of the two client classes. Cleaners soon inspected 'choosy' plates before 'resident' plates. This supports previous field observations that suggest that client species with access to several cleaners exert choice to receive better(immediate) service.

title: Competition encourages cooperation: client fish receive higher-quality service when cleaner fish compete
abstract: Partner choice is likely to be an important mechanism for the maintenance of cooperation in many mutualisms, and cleaner mutualisms among fish have been a model system for testing predictions of partner choice theories. Fish seeking to be cleaned (clients) face two potential problems: cleaners may cheat them by feeding on tissue or mucous, and they may have to wait for service from a cleaner. Previous work on interactions between the bluestreak cleaner wrasse, Labroides dimidiatus, and its clients indicates that cleaners provide higher-quality service to species that have the option of switching partners. These results have been interpreted as evidence that partner choice leads cleaners to behave cooperatively; however, alternative explanations exist for these between-species patterns. Here I focus on interactions between the bluestreak cleaner wrasse and a single client species that varies in its ability to switch partners to test the prediction that clients receive higher-quality service when cleaners have to compete for access to them. The results indicate that cleaners give ornate butterflyfish, Chaetodon ornatissimus, priority of access to their services when these clients can switch partners; however, ornate butterflyfish are not cheated less often when they have access to multiple cleaner stations, and they frequently respond to cheating by aggressively chasing cleaners. In combination with previous work, these results indicate that partner choice influences cooperative interactions between bluestreak cleaner wrasse and their clients, but that some clients with the ability to switch partners may use other strategies (i.e. punishment) to limit cheating by cleaners.

title: Choosy reef fish select cleaner fish that provide high-quality service
abstract: Reef fish that actively visit cleaner fish to have parasites and dead or infected tissue removed face two potential problems: they might have to wait while cleaners inspect other clients, and cleaners might feed on healthy body tissue, a behaviour that is referred to as cheating. Individuals of some ‘client’ species have large home ranges, which cover several cleaning stations, while others have small territories or home ranges with access to only one cleaning station. The former can thus choose between cleaners, while the latter cannot. We investigated whether clients with large home ranges change cleaning partners to outplay cleaners against each other to achieve (1) priority of access over clients with no choice at cleaning stations and (2) control over cheating by cleaners. We followed individuals of longnosed parrotfish, Hipposcarus harid , for up to 120 min in their natural environment and noted their interactions with cleaner wrasses, Labroides dimidiatus . Individuals were likely to return to the same cleaning station if the previous interaction had ended without conflict but changed cleaners for the next inspection if they had been either cheated or ignored, at least if the time between two consecutive visits was short. The overall attractiveness of a cleaning station seemed to be largely independent of service quality, which appeared to be similar at all stations. This is the first empirical evidence that the option to change partners is used as a control mechanism to stabilize cooperative behaviour.

title: Grooming Reciprocity in Female Tibetan Macaques Macaca Thibetana
abstract: Grooming among nonhuman primates is widespread and may represent an important service commodity that is exchanged within a biological marketplace. In this study, using focal animal sampling methods, we recorded grooming relationships among 12 adult females in a free‐ranging group of Tibetan macaques (Macaca thibetana) at Huangshan, China, to determine the influence of rank and kinship on grooming relationships, and whether females act as reciprocal traders (exchange grooming received for grooming given) or interchange traders (interchange grooming for social tolerance or other commodities). The results showed that: (1) grooming given was positively correlated with grooming received; (2) kinship did not exert a significant influence on grooming reciprocity; and (3) grooming reciprocity occurred principally between individuals of adjacent rank; however, when females of different rank groomed, females tended to groom up the hierarchy (lower ranking individuals groomed higher ranking individuals more than vice versa). Our results support the contention that both grooming reciprocity and the interchange of grooming for tolerance represent important social tactics used by female Tibetan macaques.  2012.

title: Payment for sex in a macaque mating market
abstract: In primate sexual relationships, males and females can cooperate through social trade. Market-like trading of sexual activity has been theorized, but no data have yet been presented that clearly show its existence. I collected data to test whether biological market theory could account for exchanges of male-to-female grooming and sexual activity in longtailed macaques. I explored male-to-female grooming, rates of sexual activity, and groomingemating interchanges, which were male-to-female grooming bouts that directly involved mating. Male-to-female grooming mainly occurred when females were sexually active, and males groomed females longer per bout when mating, inspection, or presentation of female hindquarters was involved. Moreover, male-to-female grooming was associated with an increase in female rates for all forms of sexual activity, where in contrast, female-to-male grooming was associated with decreased rates of mating in the groomed males. Males did not preferentially mate with swollen females or invest more grooming in them during groomingemating interchanges, as swellings did not seem to be a reliable indicator of female fertility. Rank status was correlated with grooming payment during groomingemating interchanges in favour of higher-ranked males and females. In support of a biological market interpretation, the amount of grooming a male performed on a female during groomingemating interchanges was related to the current supply of females around the interaction. The results provided evidence of a groomingemating trade that was influenced by a mating market.

title: Chacma baboon mating markets: competitor suppression mediates the potential for intersexual exchange
abstract: Contrary to the expectations of biological market theory, in species where sexual coercion is effective males often exchange resources or services with females for the opportunity to mate. This suggests that an ability to control mating partners does not preclude the need for their cooperation. We argue that this is because, in many systems, female resistance to coercion can precipitate intermale competition and, as such, a male’s mating strategy may often be better served by securing female compliance through affiliative rather than agonistic means. Based on this reasoning, we predicted that the need for males to exchange resources/services for mating access with females will be absent only when intermale power differentials are such that dominant individuals can secure uncontested access to receptive females. Accordingly, data from our long-term study site revealed no support for a biological markets model of intersexual mating exchange in chacma baboons, where the mating monopolies of alpha males are near absolute. Specifically, we found that males groomed females substantially less than they were groomed by them, their propensity to groom was poorly described by measures of female fertility, male rank, and the operational sex ratio, and their mating success was not associated with their grooming effort. We further predicted that an additional consequence of the degree of competitor suppression seen in chacma would be a reversal in the direction of intersexual trade of services for mating. We found that female grooming was positively associated with the probability that they would successfully initiate a copulation. Our study strongly suggests that it is variance in competitor suppression, not partner control, that mediates the potential for and direction of intersexual cooperative exchange.  biological markets, coercion, competitor suppression, intersexual cooperation.

title: Ant-aphid mutualisms: the impact of honeydew production and honeydew sugar composition on ant preferences
abstract: The honeydew composition and production of four aphid species feeding on Tanacetum vulgare, and mutualistic relationships with the ant Lasius niger were studied. In honeydew of Metopeurum fuscoviride and Brachycaudus cardui, xylose, glucose, fructose, sucrose, maltose, melezitose, and raffinose were detected. The proportion of trisaccharides (melezitose, raffinose) ranged between 20% and 35%. No trisaccharides were found in honeydew of Aphis fabae, and honeydew of Macrosiphoniella tanacetaria consisted of only xylose, glucose and sucrose. M. fuscoviride produced by far the largest amounts of honeydew per time unit (880 μg/aphid per hour), followed by B. cardui (223 μg/aphid per hour), A. fabae (133 μg/aphid per hour) and M. tanacetaria (46 μg/aphid per hour). The qualitative and quantitative honeydew production of the aphid species corresponded well with the observed attendance by L. niger. L. niger workers preferred trisaccharides over disaccharides and monosaccharides when these sugars were offered in choice tests. The results are consistent with the ants' preference for M. fuscoviride, which produced the largest amount of honeydew including a considerable proportion of the trisaccharides melezitose and raffinose. The preference of L. niger for B. cardui over A. fabae, both producing similar amounts of honeydew, may be explained by the presence of trisaccharides and the higher total sugar concentration in B. cardui honeydew. M. tanacetaria, which produced only low quantities of honeydew with no trisaccharides was not attended at all by L. niger.

title: ECOLOGY AND EVOLUTION OF APHID-ANT INTERACTIONS
abstract: ▪ Abstract Aphids and ants are two abundant and highly successful insect groups, which often live in the same habitat and therefore are likely to interact with one another. Whether the outcome of such an interaction is a predator-prey or mutualistic one is dependent on what each partner has to offer relative to the needs of the other. Consequently, understanding why some aphids enter mutualistic interactions with ants is dependent on understanding the physiological, ecological, and evolutionary traits of both partners. This includes an appreciation of the spatial, temporal, and taxonomic context in which mutualistic interactions developed. In this review, we use aphid-ant interactions to illustrate the whole range of interactions from antagonistic to mutualistic as well as to identify the processes affecting the degree of association and in particular the context within which such interactions evolved. The constraints of establishing and maintaining beneficial interactions between aphids and ants is addre...

title: Market effects offset the reciprocation of grooming in free-ranging redfronted lemurs, Eulemur fulvus rufus

abstract: Social grooming is a commonly observed affiliative behaviour in primates. Grooming has been suggested to represent a service in a biological marketplace, exchanged either for grooming or for other social commodities or services. Accordingly, grooming is predicted to be approximately reciprocated within a dyad when no other services are being exchanged, but it should be more asymmetrical if partners have different quantities of other services to offer. We analysed 412 grooming bouts observed in four groups of free-ranging redfronted lemurs to test this prediction. Grooming in this species seems to take place in a highly reciprocal manner because partners usually alternate in the roles of groomer and gromee within a grooming bout. However, within dyads there were asymmetries in the duration of grooming given and received. In both sexes, more grooming was directed from low-ranking towards high-ranking individuals than vice versa, and in males this asymmetry became more pronounced as the number of subordinates per group increased. Grooming in bisexual dyads was generally skewed in favour of males, but patterns of grooming between the sexes were less clear than within the sexes. In addition, aggression occurred at high frequencies between classes of individuals that were characterized by nonreciprocal grooming, suggesting that grooming may serve as a means to reduce aggression in dyads with a high potential for conflicts. Taken together, our results indicate that a strict reciprocation of grooming can be offset by power differentials between partners, where grooming appears to be traded by subordinates in exchange for the tolerance of dominants.

title: When females trade grooming for grooming: testing partner control and partner choice models of cooperation in two primate species
abstract: We tested predictions following from the biological market paradigm using reciprocated grooming sessions among the adult females in a sooty mangabey, Cercocebus atys, group with 35 females (Ivory Coast) and in two groups of vervet monkeys, Chlorocebus aethiops (South Africa) with four and seven females, respectively. Closely ranked females often groomed frequently. The exchanges within such dyads were generally characterized by time matching, but the subordinates groomed for longer than their dominant partners. The reciprocal nature of over 90% of the grooming sessions allowed us to investigate ‘partner control’ strategies such as ‘parcelling’ and ‘raising the stakes’. Females of both species neither parcelled nor gradually invested more grooming in the course of sessions. Rather, the longer bouts of a grooming session were usually at the beginning of the session and the length of the first bout reliably predicted the length of the whole session for frequently grooming partners. Furthermore, we compared potential trust-building behaviour (or ‘strategies’) in frequent and infrequent grooming partners. We found that infrequent groomers of both species showed no signs of trust building and that the first bout they invested in a grooming session did not predict the session length. We conclude that each female has a good knowledge of her value as a grooming partner within each dyad and knows how much she has to invest to receive a satisfactory amount of grooming within the same session.

title: Grooming and the value of social relationships in cooperatively breeding meerkats
abstract: In social mammals, grooming is used not only for hygienic purposes; it also serves social functions. We examined the grooming patterns of dominant males in groups of cooperatively breeding meerkats, Suricata suricatta, to test the hypothesis that the distribution of grooming reflects the value of social relationships. Grooming between dominant individuals was the most commonly observed interaction. Grooming by the dominant male was less common when the dominant female was in oestrus. Grooming between dominant males and subordinates was asymmetrical, with dominant males receiving more grooming and providing less grooming to subordinates. Because subordinate males defend the group against extragroup males, the dominant male was predicted to groom subordinate males more often than to groom subordinate females. Although the sex of a subordinate did not affect the probability of grooming occurring, dominant males groomed subordinate males for longer than they groomed subordinate females. The duration of grooming by a dominant male was longer in small groups than large groups, suggesting that grooming activity is constrained by time. In contrast, subordinates groomed dominant males for longer in larger groups, and as a result, the asymmetry of grooming exchanges was exaggerated in larger groups. This result is consistent with the biological market theory, which predicts that the value of subordinates is dependent on group size and that subordinates will alter grooming efforts accordingly. These results suggest that meerkat grooming serves several social functions, such as the maintenance of valuable and/or fitness-enhancing relationships.

title: Negotiation, Sanctions, and Context Dependency in the Legume-Rhizobium Mutualism
abstract: Two important questions about mutualisms are how the fitness costs and benefits to the mutualist partners are determined and how these mechanisms affect the evolutionary dynamics of the mutualism. We tackle these questions with a model of the legume-rhizobium symbiosis that regards the mutualism outcome as a result of biochemical negotiations between the plant and its nodules. We explore the fitness consequences of this mechanism to the plant and rhizobia and obtain four main results. First, negotiations permit the plant to differentially reward more-cooperative rhizobia—a phenomenon termed “plant sanctions”—but only when more-cooperative rhizobia also provide the plant with good outside options during negotiations with other nodules. Second, negotiations may result in seemingly paradoxical cases where the plant is worse off when it has a “choice” between two strains of rhizobia than when infected by either strain alone. Third, even when sanctions are effective, they are by themselves not sufficient to maintain cooperative rhizobia in a population: less cooperative strains always have an advantage at the population level. Finally, partner fidelity feedback, together with genetic correlations between a rhizobium strain’s cooperativeness and the outside options it provides, can maintain cooperative rhizobia. Our results show how joint control over the outcome of a mutualism through the proximate mechanism of negotiation can affect the evolutionary dynamics of interspecific cooperation.

title: Expanding comparative–advantage biological market models: contingency of mutualism on partner's resource requirements and acquisition trade–offs
abstract: We expand the comparative–advantage biological market–modelling framework to show how differences between partners, both in their abilities to acquire two resources and in their requirements for those resources, can affect the net benefit of participating in interspecific resource exchange. In addition, the benefits derived from resource trading depend strongly on the nature of the trade–off between the acquisition of one resource and the acquisition of another, described here by the shape (linear, convex or concave) of the resource acquisition constraints of the individuals involved. Combined with previous results, these analyses provide a suite of predictions about whether or not resource exchange is beneficial for two heterospecific individuals relative to a strategy of non–interaction. The benefit derived from resource exchange depends on three factors: (i) relative differences between the partners in their resource acquisition abilities; (ii) relative differences between the partners in their resource requirements; and (iii) variation in the shape of resource acquisition trade–offs. We find that such an explicit consideration of resource requirements and acquisition abilities can provide useful and sometimes non–intuitive predictions about the benefits of resource exchange, and also which resources should be traded by which species.

title: Honeydew production and honeydew sugar composition of polyphagous black bean aphid, Aphis fabae (Hemiptera: Aphididae) on various host plants and implications for ant-attendance
abstract: The black bean aphid, Aphis fabae, is polyphagous and its life cycle involves seasonal migration between summer and winter host plants. The aphids are regularly tended by honeydew-collecting ants. This study investigates whether differences exist in honeydew production and honeydew sugar composition for A. fabae subspecies feeding on various host plants and whether such dif- ferences reflect differences in the intensity of ant-atttendance ( Lasius niger). A. f. fabae feeding on the perennial summer host, Tanacetum vulgare, produced twice the amount of honeydew (ca. 110 µg per aphid . h -1 ) than when feeding on the annual host plants Vicia faba or Chenopodium album. Honeydew production of A. f. cirsiia- canthoides feeding on the creeping thistle Cirsium arvense was the highest measured in this study (ca. 150 µg per aphid . h -1 ). Total sugar concentration in the honeydew of A. f. fabae did not differ when feeding on various summer hosts, whereas the honeydew of A. f. cirsiiacanthoides on C. arvense contained a significantly higher amount of total sugars. The trisaccharide melezitose was the dominant sugar in all the honeydew samples, except for the honeydew of A. f. fabae and A. f. evonymi feeding on the woody winter host plant Evonymus europaeus. The highest proportion of melezitose (80% of total sugars) was found in the honeydew of A. f. cirsi- iacanthoides feeding on C. arvense. In this subspecies, the intensity of ant-attendance was also highest. The results confirm our hypothesis, that the sugar richness of the honeydew (rate of honeydew secretion × total sugar concentration) along with the presence of the attractant sugar melezitose are the critical factors in determining the extent of ant-attendance. In A. f. fabae feeding on the spindle tree E. europaeus, the total sugar concentration of the honeydew as well as the sugar composi- tion differed significantly between generations.

title: Mating First, Mating More: Biological Market Fluctuation in a Wild Prosimian
abstract: In biology, economics, and politics, distributive power is the key for understanding asymmetrical relationships and it can be obtained by force (dominance) or trading (leverage). Whenever males cannot use force, they largely depend on females for breeding opportunities and the balance of power tilts in favour of females. Thus, males are expected not only to compete within their sex-class but also to exchange services with the opposite sex. Does this mating market, described for humans and apes, apply also to prosimians, the most ancestral primate group? To answer the question, we studied a scent-oriented and gregarious lemur, Propithecus verreauxi (sifaka), showing female dominance, promiscuous mating, and seasonal breeding. We collected 57 copulations involving 8 males and 4 females in the wild (Berenty Reserve, South Madagascar), and data (all occurrences) on grooming, aggressions, and marking behaviour. We performed the analyses via exact Spearman and matrix correlations. Male mating priority rank correlated with the frequency of male countermarking over female scents but not with the proportion of fights won by males over females. Thus, males competed in an olfactory tournament more than in an arena of aggressive encounters. The copulation frequency correlated neither with the proportion of fights won by males nor with the frequency of male countermarking on female scents. Male-to-female grooming correlated with female-to-male grooming only during premating. Instead, in the mating period male-to-female grooming correlated with the copulation frequency. In short, the biological market underwent seasonal fluctuations, since males bargained grooming for sex in the mating days and grooming for itself in the premating period. Top scent-releasers gained mating priority (they mated first) and top groomers ensured a higher number of renewed copulations (they mated more). In conclusion, males maximize their reproduction probability by adopting a double tactic and by following market fluctuations.

title: Variation and the response to variation as a basis for successful cooperation
abstract: In applying game theory to problems in biology, differences between individuals are often ignored. In particular, when analysing the evolution of cooperation it is often implicitly assumed that ignoring variation will produce predictions that approximate the solution when differences are included. This need not be true. As we demonstrate, differences are not innocuous noise, but can fundamentally change the nature of a game. Even small amounts of variability can stabilize cooperation by, for example, maintaining the need to deal with cheaters. Differences promote the need to learn about others in an interaction, leading to contingent behaviour that can reduce conflict, and to negotiated outcomes that may or may not be more cooperative than unconditional actions. Once there are mechanisms such as mutation and environmental influences that maintain variation within populations, whether cooperation evolves may depend on the variation in the cooperativeness trait. Variation means that it may be worth taking a chance that a partner is cooperative by being cooperative. When there are markets, so that individuals can break off interactions to seek a better partner, variation promotes choosiness and hence penalizes those uncooperative individuals, who are rejected. Variation promotes the need to monitor the previous behaviour of others, and once this social sensitivity exists, the need to maintain a good reputation can promote cooperation.

title: Supply and demand determine the market value of food providers in wild vervet monkeys
abstract: Animals neither negotiate verbally nor conclude binding contracts, but nevertheless regularly exchange goods and services without overt coercion and manage to arrive at agreements over exchange rates. Biological market theory predicts that such exchange rates fluctuate according to the law of supply and demand. Previous studies showed that primates pay more when commodities become scarcer: subordinates groomed dominants longer before being tolerated at food sites in periods of shortage; females groomed mothers longer before obtaining permission to handle their infants when there were fewer newborns and males groomed fertile females longer before obtaining their compliance when fewer such females were present. We further substantiated these results by conducting a 2-step experiment in 2 groups of free-ranging vervet monkeys in the Loskop Dam Nature Reserve, South Africa. We first allowed a single low-ranking female to repeatedly provide food to her entire group by triggering the opening of a container and measured grooming bouts involving this female in the hour after she made the reward available. We then measured the shifts in grooming patterns after we added a second food container that could be opened by another low-ranking female, the second provider. All 4 providers received more grooming, relative to the amount of grooming they provided themselves. As biological market theory predicts, the initial gain of first providers was partially lost again after the introduction of a second provider in both groups. We conclude that grooming was fine-tuned to changes in the value of these females as social partners.

title: Host sanctions and the legume–rhizobium mutualism
abstract: Explaining mutualistic cooperation between species remains one of the greatest problems for evolutionary biology. Why do symbionts provide costly services to a host, indirectly benefiting competitors sharing the same individual host? Host monitoring of symbiont performance and the imposition of sanctions on ‘cheats’ could stabilize mutualism. Here we show that soybeans penalize rhizobia that fail to fix N2 inside their root nodules. We prevented a normally mutualistic rhizobium strain from cooperating (fixing N2) by replacing air with an N2-free atmosphere (Ar:O2). A series of experiments at three spatial scales (whole plants, half root systems and individual nodules) demonstrated that forcing non-cooperation (analogous to cheating) decreased the reproductive success of rhizobia by about 50%. Non-invasive monitoring implicated decreased O2 supply as a possible mechanism for sanctions against cheating rhizobia. More generally, such sanctions by one or both partners may be important in stabilizing a wide range of mutualistic symbioses.

title: Failure to fix nitrogen by non-reproductive symbiotic rhizobia triggers host sanctions that reduce fitness of their reproductive clonemates
abstract: The legume–rhizobia symbiosis is a classical mutualism where fixed carbon and nitrogen are exchanged between the species. Nonetheless, the plant carbon that fuels nitrogen (N2) fixation could be diverted to rhizobial reproduction by ‘cheaters’—rhizobial strains that fix less N2 but potentially gain the benefit of fixation by other rhizobia. Host sanctions can decrease the relative fitness of less-beneficial reproductive bacteroids and prevent cheaters from breaking down the mutualism. However, in certain legume species, only undifferentiated rhizobia reproduce, while only terminally differentiated rhizobial bacteroids fix nitrogen. Sanctions were, therefore, tested in two legume species that host non-reproductive bacteroids. We demonstrate that even legume species that host non-reproductive bacteroids, specifically pea and alfalfa, can severely sanction undifferentiated rhizobia when bacteroids within the same nodule fail to fix N2. Hence, host sanctions by a diverse set of legumes play a role in maintaining N2 fixation.

title: Preferential allocation to beneficial symbiont with spatial structure maintains mycorrhizal mutualism.
abstract: Mutualisms, beneficial interactions between species, are expected to be unstable because delivery of benefit likely involves fitness costs and selection should favour partners that deliver less benefit. Yet, mutualisms are common and persistent, even in the largely promiscuous associations between plants and soil microorganisms such as arbuscular mycorrhizal fungi. In two different systems, we demonstrate preferential allocation of photosynthate by host plants to the more beneficial of two AM fungal symbionts. This preferential allocation could allow the persistence of the mutualism if it confers sufficient advantage to the beneficial symbiont that it overcomes the cost of mutualism. We find that the beneficial fungus does increase in biomass when the fungi are spatially separated within the root system. However, in well-mixed fungal communities, non-beneficial fungi proliferate as expected from their reduced cost of mutualism. Our findings suggest that preferential allocation within spatially structured microbial communities can stabilize mutualisms between plants and root symbionts.

title: Reciprocal Rewards Stabilize Cooperation in the Mycorrhizal Symbiosis
abstract: Plants and their associated fungi reward partners that offer the best resources to sustain mutualism in complex systems. Plants and their arbuscular mycorrhizal fungal symbionts interact in complex underground networks involving multiple partners. This increases the potential for exploitation and defection by individuals, raising the question of how partners maintain a fair, two-way transfer of resources. We manipulated cooperation in plants and fungal partners to show that plants can detect, discriminate, and reward the best fungal partners with more carbohydrates. In turn, their fungal partners enforce cooperation by increasing nutrient transfer only to those roots providing more carbohydrates. On the basis of these observations we conclude that, unlike many other mutualisms, the symbiont cannot be “enslaved.” Rather, the mutualism is evolutionarily stable because control is bidirectional, and partners offering the best rate of exchange are rewarded.

title: Patterns of below-ground plant interconnections established by means of arbuscular mycorrhizal networks.
abstract: The ability of arbuscular mycorrhizal (AM) networks originating from plants of different species, genera and families to become interconnected by means of hyphal anastomoses was assessed. An in vivo two-dimensional experimental model system was used to reveal the occurrence of linkages between contiguous mycorrhizal networks spreading from Allium porrum root systems and those originating from Daucus carota, Gossypium hirsutum, Lactuca sativa, Solanum melongena, colonized by Glomus mosseae. Percentages of hyphal contacts leading to anastomosis between extraradical networks originating from different plant species ranged from 44% in the pairing A. porrum-S. melongena to 49% in A. porrum-G. hirsutum. DAPI and Sytox stainings detected nuclei in the middle of fusion bridges connecting different mycorrhizal networks. Present data suggest that, by means of anastomoses, AM fungal mycelium would potentially create an indefinitely large network interconnecting different plants in a community, and that, in the absence of sexual recombination, the intermingling of nuclei in extraradical mycelium may provide endless opportunities for the exchange of genetic material.

title: Underground resource allocation between individual networks of mycorrhizal fungi.
abstract: Fusions between individual mycelia of arbuscular mycorrhizal (AM) fungi have been observed in two-dimensional systems but never in soil systems. Here, phosphorus ((32)P) labelling was used to demonstrate nutrient transfer between individual mycelia and to investigate the possible role of anastomosis. Trifolium subterraneum colonized by Glomus mosseae were grown in root-retaining mesh bags, which were placed 20 cm apart. The mycelium of one plant, the donor, had access to (32)P-labelled soil placed adjacent to the mesh bag. Transfer of (32)P from the donor mycelium to the receiver plant was measured at three harvests. In a second-harvest control treatment the receiver was colonized by Glomus caledonium in order to determine whether transfer occurred by other means than hyphal fusions. Significant amounts of P were transferred to the receiver plant at the last harvests when the two mycelia of G. mosseae overlapped. The transfer probably occurred via anastomoses between the mycelia as no transfer of (32)P was detected between the mycelia of different fungi at the second harvest. The indicated ability of AM fungal mycelia to anastomose in soil has implications for the formation of large plant-interlinking functional networks, long-distance nutrient transport and retention of nutrients in readily plant-available pools.

title: Plants as resource islands and storage units--adopting the mycocentric view of arbuscular mycorrhizal networks.
abstract: The majority of herbaceous plants are connected by arbuscular mycorrhizal (AM) fungi in complex networks, but how this affects carbon (C) and phosphorus (P) allocation among symbionts is poorly understood. We utilized a monoxenic AM system where hyphae from donor roots colonized two younger receiver roots of varying C status. AM fungal C allocation from donor to receiver compartments was followed by measuring the (13)C contents in fungal- and plant-specific lipids, and P movement from a hyphal compartment was traced using (33)P. Four times more (13)C was translocated from donor to C-limited receiver roots, but C remained in fungal tissue. Root C status did not influence the overall AM colonization, but arbuscule density was twice as high in non-C-limited roots, and they received 10 times more (33)P. The number of hyphal connections between compartments did not influence C and P allocation. Interestingly, there were more fungal storage lipids, but fewer structural lipids inside C-limited roots. Our results indicate that AM colonization may poorly reflect host quality as C can be supplied from neighboring roots. A mycocentric view of the symbiosis is proposed where C-delivering hosts are resource islands for the exchange of P for C, and C-limited hosts are storage units.

title: Cooperation and its Evolution
abstract: This collection reports on the latest research on an increasingly pivotal issue for evolutionary biology: cooperation. The chapters are written from a variety of disciplinary perspectives and utilize research tools that range from empirical survey to conceptual modeling, reflecting the rich diversity of work in the field. They explore a wide taxonomic range, concentrating on bacteria, social insects, and, especially, humans. Part I ("Agents and Environments") investigates the connections of social cooperation in social organizations to the conditions that make cooperation profitable and stable, focusing on the interactions of agent, population, and environment. Part II ("Agents and Mechanisms") focuses on how proximate mechanisms emerge and operate in the evolutionary process and how they shape evolutionary trajectories. Throughout the book, certain themes emerge that demonstrate the ubiquity of questions regarding cooperation in evolutionary biology: the generation and division of the profits of cooperation; transitions in individuality; levels of selection, from gene to organism; and the "human cooperation explosion" that makes our own social behavior particularly puzzling from an evolutionary perspective.

title: Partner choice in Medicago Truncatula–Sinorhizobium symbiosis
abstract: In nitrogen-fixing symbiosis, plant sanctions against ineffective bacteria have been demonstrated in previous studies performed on soybean and yellow bush lupin, both developing determinate nodules with Bradyrhizobium sp. strains. In this study, we focused on the widely studied symbiotic association Medicago truncatula–Sinorhizobium meliloti, which forms indeterminate nodules. Using two strains isolated from the same soil and displaying different nitrogen fixation phenotypes on the same fixed plant line, we analysed the existence of both partner choice and plant sanctions by performing split-root experiments. By measuring different parameters such as the nodule number, the nodule biomass per nodule and the number of viable rhizobia per nodule, we showed that M. truncatula is able to select rhizobia based on recognition signals, both before and after the nitrogen fixation step. However, no sanction mechanism, described as a decrease in rhizobia fitness inside the nodules, was detected. Consequently, even if partner choice seems to be widespread among legumes, sanction of non-effective rhizobia might not be universal.

title: Carbon availability triggers fungal nitrogen uptake and transport in arbuscular mycorrhizal symbiosis
abstract: The arbuscular mycorrhizal (AM) symbiosis, formed between the majority of land plants and ubiquitous soil fungi of the phylum Glomeromycota, is responsible for massive nutrient transfer and global carbon sequestration. AM fungi take up nutrients from the soil and exchange them against photosynthetically fixed carbon (C) from the host. Recent studies have demonstrated that reciprocal reward strategies by plant and fungal partners guarantee a “fair trade” of phosphorus against C between partners [Kiers ET, et al. (2011) ], but whether a similar reward mechanism also controls nitrogen (N) flux in the AM symbiosis is not known. Using mycorrhizal root organ cultures, we manipulated the C supply to the host and fungus and followed the uptake and transport of N sources in the AM symbiosis, the enzymatic activities of arginase and urease, and fungal gene expression in the extraradical and intraradical mycelium. We found that the C supply of the host plant triggers the uptake and transport of N in the symbiosis, and that the increase in N transport is orchestrated by changes in fungal gene expression. N transport in the symbiosis is stimulated only when the C is delivered by the host across the mycorrhizal interface, not when C is supplied directly to the fungal extraradical mycelium in the form of acetate. These findings support the importance of C flux from the root to the fungus as a key trigger for N uptake and transport and provide insight into the N transport regulation in the AM symbiosis.

title: Spatial polymorphism of bacteriocins and other allelopathic traits
abstract: SummaryMany bacterial species carry plasmids that encode both the production of a highly specific toxin (bacteriocin) that kills competitors of the same species and immunity to that toxin. A great diversity of bacteriocins is produced within a single species. I present a model for the dynamics of competition between allelopathic and susceptible types. The model applies to most kinds of allelopathic competition. My primary goal is, however, to explain the widespread genetic polymorphism for bacteriocins. The model includes competition for scarce resources, competition through toxin production, spatial diffusion of individuals and toxins at varying rates and spatial variation in habitat quality. I draw three main conclusions from this ‘reaction—diffusion’ model. (1) Polymorphism of toxin producers and susceptibles cannot be maintained within a single spatial location when individuals and the toxin mix randomly. (2) Susceptibles are generally favoured in poor habitats, where the rate of resource competition per interaction increases relative to the resource-independent rate of toxic killing. By contrast, toxic producers are generally favoured in good habitats, where the rate of resource competition is lower. (3) Spatial variation in habitat quality can lead to spatial polymorphism; susceptibles tend to win in poor habitats and producers tend to win in good habitats.

title: Self-structuring in spatial evolutionary ecology.
abstract: Spatial self-structuring has been a focus of recent interest among evolutionary ecologists. We review recent developments in the study of the interplay between spatial self-structuring and evolution. We first discuss the relative merits of the various theoretical approaches to spatial modelling in ecology. Second, we synthesize the main theoretical studies of the evolution of cooperation in spatially structured populations. We show that population viscosity is generally beneficial to cooperation, because cooperators can reap additional benefits from being clustered. A similar mechanism can explain the evolution of honest communication and of reduced virulence in host-parasite interactions. We also discuss some recent innovative empirical results that test these theories. Third, we show the relevance of these results to the general field of evolutionary ecology. An important conclusion is that kin selection is the main process that drives evolution of cooperation in viscous populations. Many results of kin selection theory can be recovered as emergent properties of spatial ecological dynamics. We discuss the implications of these results for the study of multilevel selection and evolutionary transitions. We conclude by sketching some perspectives for future research, with a particular emphasis on the topics of evolutionary branching, criticality, spatial fluctuations and experimental tests of theoretical predictions. Space is the place - Sun Ra.

title: Dynamics in the mixed microbial concourse.
abstract: Isolated, clonal populations of cells are rarely found in nature. The emergent properties of microbial consortia present a challenge for the systems approach to biology, as chances for competition, communication, or collaboration multiply with the number of interacting agents. This review focuses on recent work on intercourse within biofilms, among quorum-sensing populations, and between cross-feeding metabolic cooperators. New tools from synthetic biology allow microbial interactions to be designed and tightly controlled, creating valuable model systems. We address both natural and synthetic partnerships, with an emphasis on how system behaviors derive from the properties of their components. Essential features of microbial biology arose in the context of a very mixed culture and cannot be understood without unscrambling it.

title: NOVEL COOPERATION EXPERIMENTALLY EVOLVED BETWEEN SPECIES
abstract: Cooperation violates the view of “nature red in tooth and claw” that prevails in our understanding of evolution, yet examples of cooperation abound. Most work has focused on maintenance of cooperation within a single species through mechanisms such as kin selection. The factors necessary for the evolutionary origin of aiding unrelated individuals such as members of another species have not been experimentally tested. Here, I demonstrate that cooperation between species can be evolved in the laboratory if (1) there is preexisting reciprocation or feedback for cooperation, and (2) reciprocation is preferentially received by cooperative genotypes. I used a two species system involving Salmonella enterica ser. Typhimurium and an Escherichia coli mutant unable to synthesize an essential amino acid. In lactose media Salmonella consumes metabolic waste from E. coli, thus creating a mechanism of reciprocation for cooperation. Growth in a spatially structured environment assured that the benefits of cooperation were preferentially received by cooperative genotypes. Salmonella evolved to aid E. coli by excreting a costly amino acid, however this novel cooperation disappeared if the waste consumption or spatial structure were removed. This study builds on previous work to demonstrate an experimental origin of interspecific cooperation, and to test the factors necessary for such interactions to arise.

title: Phototrophic consortia: model systems for symbiotic interrelations between prokaryotes
abstract: Most symbiotic prokaryotes known to date have been found in association with eukaryotes, whereas only few (3.5%) bacteria or archaea are known that have established interactions with other prokaryotes. As revealed by direct microscopic investigations, however, multiple morphotypes of highly structured associations of different prokaryotes exist in nature. These so-called consortia appear to represent the most developed type of bacterial interaction. Phototrophic consortia are associations of green sulfur bacteria that surround a central chemotrophic bacterium. In some natural environments, almost all cells of green sulfur bacteria occur in the associated state, i.e. as epibionts of phototrophic consortia. In contrast to earlier speculations, the central bacterium belongs to the β-Proteobacteria. Within the consortia, the green sulfur bacterial epibionts grow photolithoautotrophically, utilizing exogenous sulfide as photosynthetic electron donor. The entire consortium does not appear to be independent of organic carbon compounds, since it exhibits chemotaxis towards 2-oxoglutarate and, as demonstrated by microautoradiography, can also incorporate this compound. Intact consortia exhibit a scotophobic response in which the bacteriochlorophylls of the epibionts function as light sensors, whereas the chemotrophic central bacterium confers motility upon the association. Hence, a signal exchange must occur between the different bacteria. Based on their 16S rRNA gene sequences, the epibionts represent distinct phylotypes that are often only distantly related to known species of green sulfur bacteria. Since phototrophic consortia have recently become available in enrichment cultures, they can now serve as suitable model systems for the investigation of the molecular mechanisms of cell-cell recognition and signal exchange, and for studies of the coevolution of nonrelated prokaryotes.

title: Close Interspecies Interactions between Prokaryotes from Sulfureous Environments
abstract: Green sulfur bacteria are obligate photolithoautotrophs that require highly reducing conditions for growth and can utilize only a very limited number of carbon substrates. These bacteria thus inhabit a very narrow ecologic niche. However, several green sulfur bacteria have overcome the limits of immobility by entering into a symbiosis with motile Betaproteobacteria in a type of multicellular association termed phototrophic consortia. One of these consortia, “Chlorochromatium aggregatum,” has recently been established as the first culturable model system to elucidate the molecular basis of this symbiotic interaction. It consists of 12–20 green sulfur bacteria epibionts surrounding a central, chemoheterotrophic betaproteobacterium in a highly ordered fashion. Recent genomic, transcriptomic, and proteomic studies of “C. aggregatum” and its epibiont provide insights into the molecular basis and the origin of the stable association between the two very distantly related bacteria. While numerous genes of central metabolic pathways are upregulated during the specific symbiosis and hence involved in the interaction, only a limited number of unique putative symbiosis genes have been detected in the epibiont. Green sulfur bacteria therefore are preadapted to a symbiotic lifestyle. The metabolic coupling between the bacterial partners appears to involve amino acids and highly specific ultrastructures at the contact sites between the cells. Similarly, the interaction in the equally well studied archaeal consortia consisting of Nanoarchaeum equitans and its host Ignicoccus hospitalis is based on the transfer of amino acids while lacking the highly specialized contact sites observed in phototrophic consortia.

title: Symbiosis: mechanisms and model systems.
abstract: Foreword K. Jeon.  Symbionts Keystone Species D.P. Zook. Morphological aspects P. Nardon, H. Charles. Symbiosis and Their Consequences for Community and Applied Ecology D. Secord. II: Origin & Evolution. What's in a Tree? Does Horizontal Gene Transfer Determine Microbial Taxonomy L. Olendzenski, et al. The Commonality of Cyanobacterial Endosymbioses Does Not Support the Endosymbiotic Theory for Origin of Eukaryotic Organelles T.E. Jensen. Non Symbiotic Origin of Locomotory Organelles M. Rizzotti. The Cyanelle (Muroplast) of Cyano paradoxa: A Paradigm for Endosymbiotic Organelle Evolution W. Loffelhardt, H.J. Bohnert. The Syntrophy Hypothesis for the Origin of Eukaryotes P. Lopez-Garcia, D. Moreira. Thermodynamic Ecology of Hydrogen-based Syntrophy D.L. Valentine. Nutritional Syntrophies and Consortia as Models for the Origin of Mitochondria D.G. Searcy. Reversion of Endosymbiosis? The Case of Bleaching in Euglana J. Krajcovic et al. Symbiosomes R. Hinde, D.A. Trautman. The Abscence of Nitrogen-Fixing Organelles Due to Timing of the Nitrogen Crisis Ch.P. McKay, R. Navarro-Gonzalez.  Formation of Eukaryotic Cell Nuclei by the Symbiosis of Archaea in Bacteria T. Shinozawa, et al. III: Bacteria, Cyanobacteria & Algae.  The Microenvironment and Photosynthetic Performance of Prochloron sp. in Symbiosis with Didemnid Ascidians M. Kuhl, A.W.D. Larkum.IV: Fungi Symbiosis. Survival Strategies in Arbuscular Mycorrhizal Symbionts M. Giovannetti. Three Part Harmony -- Ascophyllum and its Symbionts D.J. Garbary, R.J. Reckert. Arbuscular Mycorrhizae Fungi and Their Endobacteria P. Bonfante, et al. V: Lichens. Origin and Evolution of Green Lichen Algae T. Friedl, D. Bhattacharya. Lichenization of the Trentepohliales R.L. Chapman, D.A. Waters.  Reflections on a Perplexing and Controversial Lichen Photobiont V. Ahmadjian.  Symbiosis in Plants. Algae Living on Trees and in Other Strange Places W. Reisser. Cycad Coralloid Roots Housing Cyanobacteria M. Grilli Caiola. Evolution of Epicholoe/Neotyphodium Endophytes and Other Clavicipitalean J.F. White, et al. The Effect of Endophytic Fungi on Host Plant Morphogenesis S. Scannerini, et al. VII: Association with Protozoa. Anaerobic Ciliates and Their Methanogenic Endosymbionts J.H.P. Hackstein, et al. Endosymbiosis of Beta-Proteobacteria in trypanosomatid Protozoa M.C.M. Motta, et al. Ectosymbiosis in Ciliated Protozoa G. Rosati.  Symbiosis Between Foraminifera and Algae J.J. Lee. Protistan-Prokaryotic Symbiosis in Deep-Sea Sulfidic Sediments K.R. Buck, J.M. Bernhard.  Symbiosis in Insects and Higher Animals. Sponge/ Algal Symbiosis: A Diversity of Associations D.A. Trautman, R. Hinde. The Taxonomy and Evolution of the Zooxanthellae-Coral Symbiosis S.Karako, et al. Algal Symbiosis in Flatworms A.M. McCoy, I. Balzer.  Fungi in Relationship with Insect and Other Arthropods R.W. Lichtwardt. Evolution of Ascomycete-Arthropoda Symbioses J.W. Spatafora. The Laboulbeniales - An Enigmatic

title: A marine microbial consortium apparently mediating anaerobic oxidation of methane
abstract: A large fraction of globally produced methane is converted to CO2 by anaerobic oxidation in marine sediments. Strong geochemical evidence for net methane consumption in anoxic sediments is based on methane profiles, radiotracer experiments and stable carbon isotope data. But the elusive microorganisms mediating this reaction have not yet been isolated, and the pathway of anaerobic oxidation of methane is insufficiently understood. Recent data suggest that certain archaea reverse the process of methanogenesis by interaction with sulphate-reducing bacteria. Here we provide microscopic evidence for a structured consortium of archaea and sulphate-reducing bacteria, which we identified by fluorescence in situ hybridization using specific 16S rRNA-targeted oligonucleotide probes. In this example of a structured archaeal-bacterial symbiosis, the archaea grow in dense aggregates of about 100 cells and are surrounded by sulphate-reducing bacteria. These aggregates were abundant in gas-hydrate-rich sediments with extremely high rates of methane-based sulphate reduction, and apparently mediate anaerobic oxidation of methane.

title: Altruism in viscous populations — an inclusive fitness model
abstract: SummaryA viscous population (Hamilton, 1964) is one in which the movement of organisms from their place of birth is relatively slow. This viscosity has two important effects: one is that local interactions tend to be among relatives, and the other is that competition for resources tends to be among relatives. The first effect tends to promote and the second to oppose the evolution of altruistic behaviour. In a simulation model of Wilsonet al. (1992) these two factors appear to exactly balance one another, thus opposing the evolution of local altruistic behaviour. Here I show, with an inclusive fitness model, that the same result holds in a patchstructured population.

title: Can altruism evolve in purely viscous populations?
abstract: SummaryLimited dispersal is often thought to facilitate the evolution of altruism by increasing the degree of relatedness among interacting individuals. Limited dispersal can have additional effects, however, such as local population regulation, that inhibits the evolution of altruism. Many models of structured populations assume that a viscous stage of the life cycle alternates with a global mixing stage, which allows the advantages of interactions among close relatives without the disadvantages of local population regulation. Here we analyse a computer simulation model of ‘pure’ population viscosity, in which offspring are always deposited close to parents and no global mixing stage exists. As expected, limited dispersal generates a high coefficient of relatedness among interacting individuals. Patches of altruists, however, are unable to ‘export’ their productivity to other regions of the landscape and are easily invaded by selfish types from neighbouring patches. Unlike models of alternating viscosity, in which high relatedness and local population regulation can be decoupled, these two opposing effects are inextricably linked in purely viscous populations, which therefore are not conducive to the evolution of altruistic traits.

title: Natural selection for costly nutrient recycling in simulated microbial metacommunities.
abstract: Recycling of essential nutrients occurs at scales from microbial communities to global biogeochemical cycles, often in association with ecological interactions in which two or more species utilise each others' metabolic by-products. However, recycling loops may be unstable; sequences of reactions leading to net recycling may be parasitised by side-reactions causing nutrient loss, while some reactions in any closed recycling loop are likely to be costly to participants. Here we examine the stability of nutrient recycling loops in an individual-based ecosystem model based on microbial functional types that differ in their metabolism. A supplied nutrient is utilised by a "source" functional type, generating a secondary nutrient that is subsequently used by two other types-a "mutualist" that regenerates the initial nutrient at a growth rate cost, and a "parasite" that produces a refractory waste product but does not incur any additional cost. The three functional types are distributed across a metacommunity in which separate patches are linked by a stochastic diffusive migration process. Regions of high mutualist abundance feature high levels of nutrient recycling and increased local population density leading to greater export of individuals, allowing the source-mutualist recycling loop to spread across the system. Individual-level selection favouring parasites is balanced by patch-level selection for high productivity, indirectly favouring mutualists due to the synergistic productivity benefits of the recycling loop they support. This suggests that multi-level selection may promote nutrient cycling and thereby help to explain the apparent ubiquity and stability of nutrient recycling in nature.

title: Auto-/heterotrophic endosymbiosis evolves in a mature stage of ecosystem development in a microcosm composed of an alga, a bacterium and a ciliate
abstract: We investigate an ecological mechanism by which endosymbiotic associations evolve, with a particular focus on the relationship between the evolution of endosymbiosis between auto- and heterotrophic organisms, and the stages of ecosystem development. For this purpose we conducted a long-term microcosm culture composed of three species, a green alga (Chlorella vulgaris), a bacterium (Escherichia coli), and a ciliated protozoan (Tetrahymena thermophila) for 3 years. During this culture T. thermophila cells harboring Chlorella cells emerged by phagocytotic uptake, and increased in frequency, reaching ca. 80-90%. This level was maintained in the late stage of ecosystem dynamics. Analysis of the ecosystem dynamics in the microcosm revealed that a complex causal process through direct/indirect interactions among ecosystem components led to reduction in dissolved O2 and food (E. coli) available to the T. thermophila, which gave a selective advantage to the organisms in the endosymbiotic association. This result suggests that the endosymbiosis evolves in a mature stage of ecosystem development, where reproduction and survival of prospective partner organisms is highly resource-limited and density-dependent, favoring efficient matter/energy transfers among participating organisms due to physical proximity. Consequently, a complex web of interactions and pathways of matter/energy flow in ecosystem evolves from an initially simple one.

title: Extracellular electron transfer via microbial nanowires
abstract: Microbes that can transfer electrons to extracellular electron acceptors, such as Fe(iii) oxides, are important in organic matter degradation and nutrient cycling in soils and sediments. Previous investigations on electron transfer to Fe(iii) have focused on the role of outer-membrane c-type cytochromes. However, some Fe(iii) reducers lack c-cytochromes. Geobacter species, which are the predominant Fe(iii) reducers in many environments, must directly contact Fe(iii) oxides to reduce them, and produce monolateral pili that were proposed, on the basis of the role of pili in other organisms, to aid in establishing contact with the Fe(iii) oxides. Here we report that a pilus-deficient mutant of Geobacter sulfurreducens could not reduce Fe(iii) oxides but could attach to them. Conducting-probe atomic force microscopy revealed that the pili were highly conductive. These results indicate that the pili of G. sulfurreducens might serve as biological nanowires, transferring electrons from the cell surface to the surface of Fe(iii) oxides. Electron transfer through pili indicates possibilities for other unique cell-surface and cell–cell interactions, and for bioengineering of novel conductive materials.

title: Direct Exchange of Electrons Within Aggregates of an Evolved Syntrophic Coculture of Anaerobic Bacteria
abstract: Wired for Life Syntrophic bacteria live on the metabolic by-products of a partner species. The exchange of the by-products accompanies a flow of electrons in the opposite direction that helps some species grow in conditions that would otherwise be unfavorable. In mixed anaerobic cultures of two related Geobacter species, Summers et al. (p. 1413) observed that one species evolved to promote the transfer of electrons directly to the other, in large aggregated cell clusters, without coupling to common anaerobic by-products such as hydrogen or formate. Selection pressures in nine parallel populations all resulted in a point mutation that truncated a protein involved in the production of small hairlike projections involved in intercellular communication—pili—and indirectly increased the expression of a c-type multiheme cytochrome responsible for extracellular electron transfer. The evolved aggregates were conductive, suggesting that the direct exchange of electrons between partner species is a possible alternative route to anaerobic syntrophy rather than interspecies hydrogen transfer; indeed, deleting a gene that encodes a hydrogenase involved in hydrogen transfer conferred a growth advantage in the cocultures. Direct cell-to-cell electron transfer occurs between two related species of bacteria. Microbial consortia that cooperatively exchange electrons play a key role in the anaerobic processing of organic matter. Interspecies hydrogen transfer is a well-documented strategy for electron exchange in dispersed laboratory cultures, but cooperative partners in natural environments often form multispecies aggregates. We found that laboratory evolution of a coculture of Geobacter metallireducens and Geobacter sulfurreducens metabolizing ethanol favored the formation of aggregates that were electrically conductive. Sequencing aggregate DNA revealed selection for a mutation that enhances the production of a c-type cytochrome involved in extracellular electron transfer and accelerates the formation of aggregates. Aggregate formation was also much faster in mutants that were deficient in interspecies hydrogen transfer, further suggesting direct interspecies electron transfer.

title: Potential for Direct Interspecies Electron Transfer in Methanogenic Wastewater Digester Aggregates
abstract: Mechanisms for electron transfer within microbial aggregates derived from an upflow anaerobic sludge blanket reactor converting brewery waste to methane were investigated in order to better understand the function of methanogenic consortia. The aggregates were electrically conductive, with conductivities 3-fold higher than the conductivities previously reported for dual-species aggregates of Geobacter species in which the two species appeared to exchange electrons via interspecies electron transfer. The temperature dependence response of the aggregate conductance was characteristic of the organic metallic-like conductance previously described for the conductive pili of Geobacter sulfurreducens and was inconsistent with electron conduction through minerals. Studies in which aggregates were incubated with high concentrations of potential electron donors demonstrated that the aggregates had no significant capacity for conversion of hydrogen to methane. The aggregates converted formate to methane but at rates too low to account for the rates at which that the aggregates syntrophically metabolized ethanol, an important component of the reactor influent. Geobacter species comprised 25% of 16S rRNA gene sequences recovered from the aggregates, suggesting that Geobacter species may have contributed to some but probably not all of the aggregate conductivity. Microorganisms most closely related to the acetate-utilizing Methanosaeta concilii accounted for more than 90% of the sequences that could be assigned to methane producers, consistent with the poor capacity for hydrogen and formate utilization. These results demonstrate for the first time that methanogenic wastewater aggregates can be electrically conductive and suggest that direct interspecies electron transfer could be an important mechanism for electron exchange in some methanogenic systems.  The conversion of waste organic matter to methane is an important bioenergy strategy, and a similar microbial metabolism of complex organic matter in anaerobic soils and sediments plays an important role in the global carbon cycle. Studies with laboratory cultures have demonstrated that hydrogen or formate can serve as an electron shuttle between the microorganisms degrading organic compounds and methanogens. However, the importance of hydrogen and formate as intermediates in the conversion of organic matter to methane in natural communities is less clear. The possibility that microorganisms within some natural methanogenic aggregates may directly exchange electrons, rather than producing hydrogen or formate as an intermediary electron carrier, is a significant paradigm shift with implications for the modeling and design of anaerobic wastewater reactors and for understanding how methanogenic communities will respond to environmental perturbations. The conversion of waste organic matter to methane is an important bioenergy strategy, and a similar microbial metabolism of complex organic matter in anaerobic soils and sediments plays an important role in the global carbon cycle. Studies with laboratory cultures have demonstrated that hydrogen or formate can serve as an electron shuttle between the microorganisms degrading organic compounds and methanogens. However, the importance of hydrogen and formate as intermediates in the conversion of organic matter to methane in natural communities is less clear. The possibility that microorganisms within some natural methanogenic aggregates may directly exchange electrons, rather than producing hydrogen or formate as an intermediary electron carrier, is a significant paradigm shift with implications for the modeling and design of anaerobic wastewater reactors and for understanding how methanogenic communities will respond to environmental perturbations.

title: Defending against parasites: fungus-growing ants combine specialized behaviours and microbial symbionts to protect their fungus gardens
abstract: Parasites influence host biology and population structure, and thus shape the evolution of their hosts. Parasites often accelerate the evolution of host defences, including direct defences such as evasion and sanitation and indirect defences such as the management of beneficial microbes that aid in the suppression or removal of pathogens. Fungus-growing ants are doubly burdened by parasites, needing to protect their crops as well as themselves from infection. We show that parasite removal from fungus gardens is more complex than previously realized. In response to infection of their fungal gardens by a specialized virulent parasite, ants gather and compress parasitic spores and hyphae in their infrabuccal pockets, then deposit the resulting pellet in piles near their gardens. We reveal that the ants' infrabuccal pocket functions as a specialized sterilization device, killing spores of the garden parasite Escovopsis. This is apparently achieved through a symbiotic association with actinomycetous bacteria in the infrabuccal pocket that produce antibiotics which inhibit Escovopsis. The use of the infrabuccal pocket as a receptacle to sequester Escovopsis, and as a location for antibiotic administration by the ants' bacterial mutualist, illustrates how the combination of behaviour and microbial symbionts can be a successful defence strategy for hosts.

title: Symbiotic bacteria on the cuticle of the leaf-cutting ant Acromyrmex subterraneus subterraneus protect workers from attack by entomopathogenic fungi
abstract: Although only discovered in 1999, the symbiotic filamentous actinobacteria present on the integument of certain species of leaf-cutting ants have been the subject of intense research. These bacteria have been shown to specifically suppress fungal garden parasites by secretion of antibiotics. However, more recently, a wider role for these bacteria has been suggested from research revealing their generalist anti-fungal activity. Here we show, for the first time, evidence for a role of these bacteria in the defence of young worker ants against a fungal entomopathogen. Experimental removal of the bacterial bio-film using an antibiotic resulted in a significant increase in susceptibility of worker ants to infection by the entomopathogenic fungus Metarhizium anisopliae. This is the first direct evidence for the advantage of maintaining a bacterial bio-film on the cuticle as a defensive strategy of the ants themselves and not exclusively for protection of the fungus garden.

title: Ecological interactions drive evolutionary loss of traits.
abstract: Loss of traits can dramatically alter the fate of species. Evidence is rapidly accumulating that the prevalence of trait loss is grossly underestimated. New findings demonstrate that traits can be lost without affecting the external phenotype, provided the lost function is compensated for by species interactions. This is important because trait loss can tighten the ecological relationship between partners, affecting the maintenance of species interactions. Here, we develop a new perspective on so-called `compensated trait loss' and how this type of trait loss may affect the evolutionary dynamics between interacting organisms. We argue that: (1) the frequency of compensated trait loss is currently underestimated because it can go unnoticed as long as ecological interactions are maintained; (2) by analysing known cases of trait loss, specific factors promoting compensated trait loss can be identified and (3) genomic sequencing is a key way forwards in detecting compensated trait loss. We present a comprehensive literature survey showing that compensated trait loss is taxonomically widespread, can involve essential traits, and often occurs as replicated evolutionary events. Despite its hidden nature, compensated trait loss is important in directing evolutionary dynamics of ecological relationships and has the potential to change facultative ecological interactions into obligatory ones.

title: Using the Acropora digitifera genome to understand coral responses to environmental change
abstract: Despite the enormous ecological and economic importance of coral reefs, the keystone organisms in their establishment, the scleractinian corals, increasingly face a range of anthropogenic challenges including ocean acidification and seawater temperature rise. To understand better the molecular mechanisms underlying coral biology, here we decoded the approximately 420-megabase genome of Acropora digitifera using next-generation sequencing technology. This genome contains approximately 23,700 gene models. Molecular phylogenetics indicate that the coral and the sea anemone Nematostella vectensis diverged approximately 500 million years ago, considerably earlier than the time over which modern corals are represented in the fossil record (∼240 million years ago). Despite the long evolutionary history of the endosymbiosis, no evidence was found for horizontal transfer of genes from symbiont to host. However, unlike several other corals, Acropora seems to lack an enzyme essential for cysteine biosynthesis, implying dependency of this coral on its symbionts for this amino acid. Corals inhabit environments where they are frequently exposed to high levels of solar radiation, and analysis of the Acropora genome data indicates that the coral host can independently carry out de novo synthesis of mycosporine-like amino acids, which are potent ultraviolet-protective compounds. In addition, the coral innate immunity repertoire is notably more complex than that of the sea anemone, indicating that some of these genes may have roles in symbiosis or coloniality. A number of genes with putative roles in calcification were identified, and several of these are restricted to corals. The coral genome provides a platform for understanding the molecular basis of symbiosis and responses to environmental changes.

title: Symbiotic diversity in marine animals: the art of harnessing chemosynthesis
abstract: Chemosynthetic symbioses between bacteria and marine invertebrates were discovered 30 years ago at hydrothermal vents on the Galapagos Rift. Remarkably, it took the discovery of these symbioses in the deep sea for scientists to realize that chemosynthetic symbioses occur worldwide in a wide range of habitats, including cold seeps, whale and wood falls, shallow-water coastal sediments and continental margins. The evolutionary success of these symbioses is evident from the wide range of animal groups that have established associations with chemosynthetic bacteria; at least seven animal phyla are known to host these symbionts. The diversity of the bacterial symbionts is equally high, and phylogenetic analyses have shown that these associations have evolved on multiple occasions by convergent evolution. This Review focuses on the diversity of chemosynthetic symbionts and their hosts, and examines the traits that have resulted in their evolutionary success.

title: Metaproteomics of a gutless marine worm and its symbiotic microbial community reveal unusual pathways for carbon and energy use
abstract: Low nutrient and energy availability has led to the evolution of numerous strategies for overcoming these limitations, of which symbiotic associations represent a key mechanism. Particularly striking are the associations between chemosynthetic bacteria and marine animals that thrive in nutrient-poor environments such as the deep sea because the symbionts allow their hosts to grow on inorganic energy and carbon sources such as sulfide and CO2. Remarkably little is known about the physiological strategies that enable chemosynthetic symbioses to colonize oligotrophic environments. In this study, we used metaproteomics and metabolomics to investigate the intricate network of metabolic interactions in the chemosynthetic association between Olavius algarvensis, a gutless marine worm, and its bacterial symbionts. We propose previously undescribed pathways for coping with energy and nutrient limitation, some of which may be widespread in both free-living and symbiotic bacteria. These pathways include (i) a pathway for symbiont assimilation of the host waste products acetate, propionate, succinate and malate; (ii) the potential use of carbon monoxide as an energy source, a substrate previously not known to play a role in marine invertebrate symbioses; (iii) the potential use of hydrogen as an energy source; (iv) the strong expression of high-affinity uptake transporters; and (v) as yet undescribed energy-efficient steps in CO2 fixation and sulfate reduction. The high expression of proteins involved in pathways for energy and carbon uptake and conservation in the O. algarvensis symbiosis indicates that the oligotrophic nature of its environment exerted a strong selective pressure in shaping these associations.

title: Mechanisms of cache retrieval in long-term hoarding birds
abstract: Food hoarding and memory have primarily been studied in two bird families, the Corvidae (crows, jays, nutcrackers, etc.) and the Paridae (tits, titmice and chickadees). In both families there are species that hoard large quantities of seeds and nuts in the autumn and depend on these stores during the winter. Caches are concealed or highly inconspicuous and the most efficient way to retrieve them is to remember the exact locations. However, a long-term memory for a large number of caches may be physiologically expensive, and especially after long retention intervals, an alternative strategy could be to retrieve caches by cheaper but less efficient methods. Very few studies have been designed to investigate the decay of the memory in birds, but both field observations and experiments point in the same direction: although long-term hoarding corvids seem to possess an accurate long-term memory, long-term hoarding parids do not appear to. I discuss possible reasons for this and suggest that differences between the families in their degree of dependence on stored food or/and size-related limitations of brain capacity may be important.

title: Ants farm subterranean aphids mostly in single clone groups - an example of prudent husbandry for carbohydrates and proteins?
abstract: BackgroundMutualistic interactions are wide-spread but the mechanisms underlying their evolutionary stability and ecological dynamics remain poorly understood. Cultivation mutualisms in which hosts consume symbionts occur in phylogenetically diverse groups, but often have symbiont monocultures for each host. This is consistent with the prediction that symbionts should avoid coexistence with other strains so that host services continue to benefit relatives, but it is less clear whether hosts should always favor monocultures and what mechanisms they might have to manipulate symbiont diversity. Few mutualisms have been studied in sufficient genetic detail to address these issues, so we decided to characterize symbiont diversity in the complex mutualism between multiple root aphid species and Lasius flavus ants. After showing elsewhere that three of these aphid species have low dispersal and mostly if not exclusively asexual reproduction, we here investigate aphid diversity within and between ant nest mounds.ResultsThe three focal species (Geoica utricularia, Forda marginata and Tetraneura ulmi) had considerable clonal diversity at the population level. Yet more than half of the ant mounds contained just a single aphid species, a significantly higher percentage than expected from a random distribution. Over 60% of these single-species mounds had a single aphid clone, and clones tended to persist across subsequent years. Whenever multiple species/clones co-occurred in the same mound, they were spatially separated with more than 95% of the aphid chambers containing individuals of a single clone.ConclusionsL. flavus “husbandry” is characterized by low aphid “livestock” diversity per colony, especially at the nest-chamber level, but it lacks the exclusive monocultures known from other cultivation mutualisms. The ants appear to eat most of the early instar aphids, so that adult aphids are unlikely to face limited phloem resources and scramble competition with other aphids. We suggest that such culling of carbohydrate-providing symbionts for protein ingestion may maintain maximal host yield per aphid while also benefitting the domesticated aphids as long as their clone-mates reproduce successfully. The cost-benefit logic of this type of polyculture husbandry has striking analogies with human farming practices based on slaughtering young animals for meat to maximize milk-production by a carefully regulated adult livestock population.

title: Tit for tat? A mycorrhizal fungus accumulates phosphorus under low plant carbon availability.
abstract: The exchange of carbohydrates and mineral nutrients in the arbuscular mycorrhizal (AM) symbiosis must be controlled by both partners in order to sustain an evolutionarily stable mutualism. Plants downregulate their carbon (C) flow to the fungus when nutrient levels are sufficient, while the mechanism controlling fungal nutrient transfer is unknown. Here, we show that the fungus accumulates nutrients when connected to a host that is of less benefit to the fungus, indicating a potential of the fungus to control the transfer of nutrients. We used a monoxenic in vitro model of root organ cultures associated with Glomus intraradices, in which we manipulated the C availability to the plant. We found that G. intraradices accumulated up to seven times more nutrients in its spores, and up to nine times more in its hyphae, when the C pool available to the associated roots was halved. The strongest effect was found for phosphorus (P), considered to be the most important nutrient in the AM symbiosis. Other elements such as potassium and chorine were also accumulated, but to a lesser extent, while no accumulation of iron or manganese was found. Our results suggest a functional linkage between C and P exchange.

title: Dictyostelium: Evolution, Cell Biology, and the Development of Multicellularity
abstract: Preface 1. A brief introduction to Dictyostelium discoideum and its relatives 2. A history of research on Dictyostelium discoideum 3. The evolutionary biology of Dictyostelium 4. The genome and genetics 5. Membranes and organelles of Dictyostelium 6. Cell motility and the cytoskeleton 7. The transition from growth to development: from starvation to self sustaining cAMP signal relay 8. Chemotaxis and aggregation 9. Differentiation and adhesion in the aggregate 10. Behavior of cells in the slug 11.  Formation and germination of spores 13. Resources References Index.

title: Primitive agriculture in a social amoeba
abstract: Agriculture has been a large part of the ecological success of humans. A handful of animals, notably the fungus-growing ants, termites and ambrosia beetles, have advanced agriculture that involves dispersal and seeding of food propagules, cultivation of the crop and sustainable harvesting. More primitive examples, which could be called husbandry because they involve fewer adaptations, include marine snails farming intertidal fungi and damselfish farming algae. Recent work has shown that microorganisms are surprisingly like animals in having sophisticated behaviours such as cooperation, communication and recognition, as well as many kinds of symbiosis. Here we show that the social amoeba Dictyostelium discoideum has a primitive farming symbiosis that includes dispersal and prudent harvesting of the crop. About one-third of wild-collected clones engage in husbandry of bacteria. Instead of consuming all bacteria in their patch, they stop feeding early and incorporate bacteria into their fruiting bodies. They then carry bacteria during spore dispersal and can seed a new food crop, which is a major advantage if edible bacteria are lacking at the new site. However, if they arrive at sites already containing appropriate bacteria, the costs of early feeding cessation are not compensated for, which may account for the dichotomous nature of this farming symbiosis. The striking convergent evolution between bacterial husbandry in social amoebas and fungus farming in social insects makes sense because multigenerational benefits of farming go to already established kin groups.

title: Structured habitats and the evolution of anticompetitor toxins in bacteria.
abstract: We demonstrate that in liquid cultures, defined in this study as a mass habitat, the outcome of competition between Escherichia coli that produce an antibacterial toxin (colicin) and sensitive E. coli is frequency dependent; the colicinogenic bacteria are at an advantage only when fairly common (frequencies in excess of 2 X 10(-2)). However, we also show that in a soft agar matrix, a structured habitat, the colicinogenic bacteria have an advantage even when initially rare (frequencies as low as 10(-6)). These contrasting outcomes are attributed to the colicinogenic bacteria's lower intrinsic growth rate relative to the sensitive bacteria and the different manner in which bacteria and resources are partitioned in the two types of habitats. Bacteria in a liquid culture exist as randomly distributed individuals and the killing of sensitive bacteria by the colicin augments the amount of resource available to the colicinogenic bacteria to an extent identical to that experienced by the surviving sensitive bacteria. On the other hand, the bacteria in a soft agar matrix exist as single-clone colonies. As the colicinogenic colonies release colicin, they kill neighboring sensitive bacteria and form an inhibition zone around themselves. By this action, they increase the concentration of resources around themselves and overcome their growth rate disadvantage. We suggest that structured habitats are more favorable for the evolution of colicinogenic bacteria.

title: Bacteriocins, spite and virulence
abstract: There has been much interest in using social evolution theory to predict the damage to a host from parasite infection, termed parasite virulence. Most of this work has focused on how high kinship between the parasites infecting a host can select for more prudent exploitation of the host, leading to a negative relationship between virulence and parasite kinship. However, it has also been shown that if parasites can cooperate to overcome the host, then high parasite kinship within hosts can select for greater cooperation and higher growth rates, hence leading to a positive relationship between virulence and parasite kinship. We examine the impact of a spiteful behaviour, chemical (bacteriocin) warfare between microbes, on the evolution of virulence, and find a new relationship: virulence is maximized when the frequency of kin among parasite's; social partners is low or high, and is minimized at intermediate values. This emphasizes how biological details can fundamentally alter the qualitative nature of theoretical predictions made by models of parasite virulence.

title: Antagonistic interactions peak at intermediate genetic distance in clinical and laboratory strains of Pseudomonas aeruginosa
abstract: BackgroundBacteria excrete costly toxins to defend their ecological niche. The evolution of such antagonistic interactions between individuals is expected to depend on both the social environment and the strength of resource competition. Antagonism is expected to be weak among highly similar genotypes because most individuals are immune to antagonistic agents and among dissimilar genotypes because these are unlikely to be competing for the same resources and antagonism should not yield much benefit. The strength of antagonism is therefore expected to peak at intermediate genetic distance.ResultsWe studied the ability of laboratory strains of Pseudomonas aeruginosa to prevent growth of 55 different clinical P. aeruginosa isolates derived from cystic fibrosis patients. Genetic distance was determined using genetic fingerprints. We found that the strength of antagonism was maximal among genotypes of intermediate genetic distance and we show that genetic distance and resource use are linked.ConclusionsOur results suggest that the importance of social interactions like antagonism may be modulated by the strength of resource competition.

title: Kin discrimination and cooperation in microbes.
abstract: Recognition of relatives is important in microbes because they perform many behaviors that have costs to the actor while benefiting neighbors. Microbes cooperate for nourishment, movement, virulence, iron acquisition, protection, quorum sensing, and production of multicellular biofilms or fruiting bodies. Helping others is evolutionarily favored if it benefits others who share genes for helping, as specified by kin selection theory. If microbes generally find themselves in clonal patches, then no special means of discrimination is necessary. Much real discrimination is actually of kinds, not kin, as in poison-antidote systems, such as bacteriocins, in which cells benefit their own kind by poisoning others, and in adhesion systems, in which cells of the same kind bind together. These behaviors can elevate kinship generally and make cooperation easier to evolve and maintain.

title: Mutualistic Fungi Control Crop Diversity in Fungus-Growing Ants
abstract: Leaf-cutting ants rear clonal fungi for food and transmit the fungi from mother to daughter colonies so that symbiont mixing and conflict, which result from competition between genetically different clones, are avoided. Here we show that despite millions of years of predominantly vertical transmission, the domesticated fungi actively reject mycelial fragments from neighboring colonies, and that the strength of these reactions are in proportion to the overall genetic difference between these symbionts. Fungal incompatibility compounds remain intact during ant digestion, so that fecal droplets, which are used for manuring newly grown fungus, elicit similar hostile reactions when applied to symbionts from other colonies. Symbiont control over new mycelial growth by manurial imprinting prevents the rearing of multiple crops in fungus gardens belonging to the same colony.

title: Leaf-cutting ant fungi produce cell wall degrading pectinase complexes reminiscent of phytopathogenic fungi
abstract: BackgroundLeaf-cutting (attine) ants use their own fecal material to manure fungus gardens, which consist of leaf material overgrown by hyphal threads of the basidiomycete fungus Leucocoprinus gongylophorus that lives in symbiosis with the ants. Previous studies have suggested that the fecal droplets contain proteins that are produced by the fungal symbiont to pass unharmed through the digestive system of the ants, so they can enhance new fungus garden growth.ResultsWe tested this hypothesis by using proteomics methods to determine the gene sequences of fecal proteins in Acromyrmex echinatior leaf-cutting ants. Seven (21%) of the 33 identified proteins were pectinolytic enzymes that originated from the fungal symbiont and which were still active in the fecal droplets produced by the ants. We show that these enzymes are found in the fecal material only when the ants had access to fungus garden food, and we used quantitative polymerase chain reaction analysis to show that the expression of six of these enzyme genes was substantially upregulated in the fungal gongylidia. These unique structures serve as food for the ants and are produced only by the evolutionarily advanced garden symbionts of higher attine ants, but not by the fungi reared by the basal lineages of this ant clade.ConclusionsPectinolytic enzymes produced in the gongylidia of the fungal symbiont are ingested but not digested by Acromyrmex leaf-cutting ants so that they end up in the fecal fluid and become mixed with new garden substrate. Substantial quantities of pectinolytic enzymes are typically found in pathogenic fungi that attack live plant tissue, where they are known to breach the cell walls to allow the fungal mycelium access to the cell contents. As the leaf-cutting ant symbionts are derived from fungal clades that decompose dead plant material, our results suggest that their pectinolytic enzymes represent secondarily evolved adaptations that are convergent to those normally found in phytopathogens.

title: THE EVOLUTION OF SPITE: POPULATION STRUCTURE AND BACTERIOCIN‐MEDIATED ANTAGONISM IN TWO NATURAL POPULATIONS OF XENORHABDUS BACTERIA
abstract: Spite occurs when an individual harms itself in the act of harming other individuals. Such behaviors were once assumed to be of limited evolutionary importance, as the conditions for the evolution of spite were thought to be too restrictive. Recent theoretical work, however, suggests that spatial population structure, which allows local competition among genotypes, could favor the evolution of spite. One of the clearest examples of spite is the costly production and release by bacteria of toxins (called bacteriocins) that can kill unrelated strains of the same species. Here, we establish the existence of spatial structure in two natural populations of bacteriocin‐producing bacteria. Specifically, relatedness decreased with increasing spatial distance between the field isolates. In addition, toxin‐mediated inhibitions were found only between isolates that were collected more than 1 m apart and that were generally less than 80% similar in their genomic fingerprints. Taken together, the results suggest that the bacteria are spatially structured, with mixing of genotypes and spiteful interactions at the boundaries between demes.

title: Spiteful Interactions in a Natural Population of the Bacterium Xenorhabdus bovienii
abstract: An individual behaves spitefully when it harms itself in the act of harming other individuals. One of the clearest potential examples of spite is the costly production and release of toxins called bacteriocins. Bacteriocins are toxins produced by bacteria that can kill closely related strains of the same species. Theoretical work has predicted that bacteriocin‐mediated interactions could play an important role in maintaining local genetic and/or species diversity, but these interactions have not been studied at biologically relevant scales in nature. Here we studied toxin production and among‐strain inhibitions in a natural population of Xenorhabdus bovienii. We found genetic differences and inhibitions between colonies that were collected only a few meters apart. These results suggest that spite exists in natural populations of bacteria.

title: Spiteful interactions between sympatric natural isolates of Xenorhabdus bovienii benefit kin and reduce virulence
abstract: Spite occurs when an individual harms itself in the act of harming others. Spiteful behaviour may be more pervasive in nature than commonly thought. One of the clearest examples of spite is the costly production and release of bacteriocins, antimicrobial toxins noted for their ability to kill conspecifics. A key question is to what extent these toxins provide a fitness advantage to kin of the producer cell, especially in natural communities. Additionally, when bacteria are involved in parasitic relationships, spiteful interactions are predicted to lower bacterial densities within a host, causing a reduction in parasite‐induced virulence. Using five sympatric, field‐collected genotypes of the insect pathogen Xenorhabdus bovienii, we experimentally demonstrate that bacteriocin production benefits kin within the host, and that it slows the mortality rate of the host. These results confirm that spite among naturally coexisting bacterial clones can be a successful kin‐selected strategy that has emergent effects on virulence.

title: Partner Choice in Nitrogen-Fixation Mutualisms of Legumes and Rhizobia1
abstract: Mutualistic interactions are widespread and obligatory for many organisms, yet their evolutionary persistence in the face of cheating is theoretically puzzling. Nutrient-acquisition symbioses between plants and soil microbes are critically important to plant evolution and ecosystem function, yet we know almost nothing about the evolutionary dynamics and mechanisms of persistence of these ancient mutualisms. Partner-choice and partner-fidelity are mechanisms for dealing with cheaters, and can theoretically allow mutualisms to persist despite cheaters. Many models of cooperative behavior assume pairwise interactions, while most plant-microbe nutrient-acquisition symbioses involve a single plant interacting with numerous microbes. Market models, in contrast, are well suited to mutualisms in which single plants attempt to conduct mutually beneficial resource exchange with multiple individuals. Market models assume that one partner chooses to trade with a subset of individuals selected from a market of potential partners. Hence, determining whether partner-choice occurs in plant-microbe mutualisms is critical to understanding the evolutionary persistence and dynamics of these symbioses. The nitrogen-fixation/carbon-fixation mutualism between leguminous plants and rhizobial bacteria is widespread, ancient, and important for ecosystem function and human nutrition. It also involves single plants interacting simultaneously with several to many bacterial partners, including ineffective (“cheating”) strains. We review the existing literature and find that this mutualism displays several elements of partner-choice, and may match the requirements of the market paradigm. We conclude by identifying profitable questions for future research.

title: Single-cell MALDI-MS as an analytical tool for studying intrapopulation metabolic heterogeneity of unicellular organisms.
abstract: Heterogeneity is a characteristic feature of all populations of living organisms. Here we make an attempt to validate a single-cell mass spectrometric method for detection of changes in metabolite levels occurring in populations of unicellular organisms. Selected metabolites involved in central metabolism (ADP, ATP, GTP, and UDP-Glucose) could readily be detected in single cells of Closterium acerosum by means of negative-mode matrix-assisted laser desorption/ionization (MALDI) mass spectrometry (MS). The analytical capabilities of this approach were characterized using standard compounds. The method was then used to study populations of individual cells with different levels of the chosen metabolites. With principal component analysis and support vector machine algorithms, it was possible to achieve a clear separation of individual C. acerosum cells in different metabolic states. This study demonstrates the suitability of mass spectrometric analysis of metabolites in single cells to measure cell-population heterogeneity.

title: Phenotypic heterogeneity in mycobacterial stringent response
abstract: BackgroundA common survival strategy of microorganisms subjected to stress involves the generation of phenotypic heterogeneity in the isogenic microbial population enabling a subset of the population to survive under stress. In a recent study, a mycobacterial population of M. smegmatis was shown to develop phenotypic heterogeneity under nutrient depletion. The observed heterogeneity is in the form of a bimodal distribution of the expression levels of the Green Fluorescent Protein (GFP) as reporter with the gfp fused to the promoter of the rel gene. The stringent response pathway is initiated in the subpopulation with high rel activity.ResultsIn the present study, we characterise quantitatively the single cell promoter activity of the three key genes, namely, mprA, sigE and rel, in the stringent response pathway with gfp as the reporter. The origin of bimodality in the GFP distribution lies in two stable expression states, i.e., bistability. We develop a theoretical model to study the dynamics of the stringent response pathway. The model incorporates a recently proposed mechanism of bistability based on positive feedback and cell growth retardation due to protein synthesis. Based on flow cytometry data, we establish that the distribution of GFP levels in the mycobacterial population at any point of time is a linear superposition of two invariant distributions, one Gaussian and the other lognormal, with only the coefficients in the linear combination depending on time. This allows us to use a binning algorithm and determine the time variation of the mean protein level, the fraction of cells in a subpopulation and also the coefficient of variation, a measure of gene expression noise.ConclusionsThe results of the theoretical model along with a comprehensive analysis of the flow cytometry data provide definitive evidence for the coexistence of two subpopulations with overlapping protein distributions.

title: Individuality in bacteria.
abstract: While traditionally microbiologists have examined bacterial behavior averaged over large populations, increasingly we are becoming aware that bacterial populations can be composed of phenotypically diverse individuals generated by a variety of mechanisms. Though the results of different mechanisms, the phenomena of bistability, persistence, variation in chemotactic response, and phase and antigenic variation are all strategies to develop population-level diversity. The understanding of individuality in bacteria requires an appreciation of their environmental and ecological context, and thus evolutionary theory regarding adaptations to time-variable environments is becoming more applicable to these problems. In particular, the application of game and information theory to bacterial individuality has addressed some interesting problems of bacterial behavior. In this review we discuss the mechanisms of generating population-level variability, and the application of evolutionary theory to problems of individuality in bacteria.

title: Erratum: Microbial individuality in the natural environment
abstract: The ISME Journal (2013) 7, 465–467; doi:10.1038/ismej.2012.131 Since the publication of this article, the author has identified an error contained within the following sentence (located on page 1): However, there is a third possibility that transcends the traditional concept of natureand nurture as determinants of the phenotype: that phenotypic diversity is produced independently of genetic for example, see environmental variation.

title: Evolutionary ecology of mycorrhizal functional diversity in agricultural systems
abstract: The root systems of most agronomic crops are colonized by diverse assemblages of arbuscular mycorrhizal fungi (AMF), varying in the functional benefits (e.g. nutrient transfer, pathogen protection, water uptake) provided to hosts. Little is known about the evolutionary processes that shape the composition of these fungal assemblages, nor is it known whether more diverse assemblages are beneficial to crop productivity. In this review we aim to identify the evolutionary selection pressures that shape AMF diversity in agricultural systems and explore whether promotion of AMF diversity can convincingly be linked to increases in agricultural productivity and/or sustainability. We then ask whether farmers can (and should) actively modify evolutionary selection pressures to increase AMF functioning. We focus on three agriculturally imposed selection regimes: tillage, fertilization, and continuous monoculture. We find that the uniform nature of these practices strongly selects for dominance of few AMF species. These species exhibit predictable, generally non‐beneficial traits, namely heavy investment in reproduction at the expense of nutrient scavenging and transfer processes that are beneficial for hosts. A number of focus‐points are given based on empirical and theoretical evidence that could be utilized to slow down negative selection pressures on AMF functioning, therein increasing crop benefit.

title: Chemotaxis in Bacteria
abstract: Motile Escherichia coli placed at one end of a capillary tube containing an energy source and oxygen migrate out into the tube in one or two bands, which are clearly visible to the naked eye and can also be demonstrated by photography, microscopy, and densitometry and by assaying for bacteria throughout the tube. The formation of two bands is not due to heterogeneity among the bacteria, since the bacteria in each band, when reused, will form two more bands. If an anaerobically utilizable energy source such as galactose is present in excess over the oxygen, the first band consumes all the oxygen and a part of the sugar and the second band uses the residual sugar anaerobically. On the other hand, if oxygen is present in excess over the sugar, the first band oxidizes all the sugar and leaves behind unused oxygen, and the second band uses up the residual oxygen to oxidize an endogenous energy source. The essence of the matter is that the bacteria create a gradient of oxygen or of an energy source, and then they move preferentially in the direction of the higher concentration of the chemical. As a consequence, bands of bacteria (or rings of bacteria in the case of agar plates) form and move out. These results show that E. coli is chemotactic toward oxygen and energy sources such as galactose, glucose, aspartic acid, threonine, or serine. The full repertoire of chemotactic responses by E. coli is no doubt greater than this, and a more complete list remains to be compiled. The studies reported here demonstrate that chemotaxis allows bacteria to find that environment which provides them with the greatest supply of energy. It is clearly an advantage for bacteria to be able to carry out chemotaxis, since by this means they can avoid unfavorable conditions and seek optimum surroundings. Finally, it is necessary to acknowledge the pioneering work of Englemann, Pfeffer, and the other late-19thcentury biologists who discovered chemotaxis in bacteria, and to point out that the studies reported here fully confirm the earlier reports of Beijerinck (4) and Sherris and his collaborators (5,6) on a band of bacteria chemotactic toward oxygen. By using a chemically defined medium instead of a complex broth, it has been possible to study this band more closely and to demonstrate in addition the occurrence of a second band of bacteria chemotactic toward an energy source. Beijerinck (4) did, in fact, sometimes observe a second band, but he did not offer an explanation for it.

title: Flagella-driven chemotaxis towards exudate components is an important trait for tomato root colonization by Pseudomonas fluorescens.
abstract: Motility is a major trait for competitive tomato root-tip colonization by Pseudomonas fluorescens. To test the hypothesis that this role of motility is based on chemotaxis toward exudate components, cheA mutants that were defective in flagella-driven chemotaxis but retained motility were constructed in four P. fluorescens strains. After inoculation of seedlings with a 1:1 mixture of wild-type and nonmotile mutants all mutants had a strongly reduced competitive root colonizing ability after 7 days of plant growth, both in a gnotobiotic sand system as well as in nonsterile potting soil. The differences were significant on all root parts and increased from root base to root tip. Significant differences at the root tip could already be detected after 2 to 3 days. These experiments show that chemotaxis is an important competitive colonization trait. The best competitive root-tip colonizer, strain WCS365, was tested for chemotaxis toward tomato root exudate and its major identified components. A chemotactic response was detected toward root exudate, some organic acids, and some amino acids from this exudate but not toward its sugars. Comparison of the minimal concentrations required for a chemotactic response with concentrations estimated for exudates suggested that malic acid and citric acid are among major chemo-attractants for P. fluorescens WCS365 cells in the tomato rhizosphere.

title: Boreal feather mosses secrete chemical signals to gain nitrogen.
abstract: The mechanistic basis of feather moss-cyanobacteria associations, a main driver of nitrogen (N) input into boreal forests, remains unknown. Here, we studied colonization by Nostoc sp. on two feather mosses that form these associations (Pleurozium schreberi and Hylocomium splendens) and two acrocarpous mosses that do not (Dicranum polysetum and Polytrichum commune). We also determined how N availability and moss reproductive stage affects colonization, and measured N transfer from cyanobacteria to mosses. The ability of mosses to induce differentiation of cyanobacterial hormogonia, and of hormogonia to then colonize mosses and re-establish a functional symbiosis was determined through microcosm experiments, microscopy and acetylene reduction assays. Nitrogen transfer between cyanobacteria and Pleurozium schreberi was monitored by secondary ion mass spectrometry (SIMS). All mosses induced hormogonia differentiation but only feather mosses were subsequently colonized. Colonization on Pleurozium schreberi was enhanced during the moss reproductive phase but impaired by elevated N. Transfer of N from cyanobacteria to their host moss was observed. Our results reveal that feather mosses likely secrete species-specific chemo-attractants when N-limited, which guide cyanobacteria towards them and from which they gain N. We conclude that this signalling is regulated by N demands of mosses, and serves as a control of N input into boreal forests.

title: Plant-animal mutualism in biological markets: evolutionary and ecological dynamics driven by non-heritable phenotypic variance.
abstract: Mutualism between plants and animals, such as in pollination and seed dispersal, is a fundamental mechanism facilitating the productivity and biodiversity of ecosystems, and it is often considered as an analog of a free-market economy. The coevolution of plant reward and animal choosiness, however, involves an apparent paradox due to incomplete information and limited mutation rates: plant rewards evolve only when animals are choosy, but choosy animals purge the heritable variations of plants, which then favors less choosy animals. Here we use a two-species mathematical model to illustrate how non-heritable phenotypic variances of plants may facilitate the coevolution of rewards and choosiness and solve the paradox with low mutation rates. We simultaneously track the ecological and evolutionary dynamics and show that the population ratio links the two processes and tunes the stable eco-evolutionary equilibrium. Numerical simulations confirm the analytic prediction with varying mutation rates (heritable variance). The efficiency of a biological market is generally suboptimal due to the information constraint and individual competition.

title: Cheating can stabilize cooperation in mutualisms
abstract: Mutualisms present a challenge for evolutionary theory. How is cooperation maintained in the face of selection for selfishness and cheating? Both theory and data suggest that partner choice, where one species preferentially directs aid to the more cooperative members of the other species, is central to cooperation in many mutualisms. However, the theory has only so far considered the evolutionary effects of partner choice on one of the species in a mutualism in isolation. Here, we investigate the co-evolution of cooperation and choice in a choosy host and its symbiont. Our model reveals that even though choice and cooperation may be initially selected, it will often be unstable. This is because choice reduces variation in the symbiont and, therefore, tends to remove the selective incentive for its own maintenance (a scenario paralleled in the lek paradox in female choice and policing in within-species cooperation). However, we also show that when variability is reintroduced into symbionts each generation, in the form of less cooperative individuals, choice is maintained. This suggests that the presence of cheaters and cheater species in many mutualisms is central to the maintenance of partner choice and, paradoxically, cooperation itself.

title: Supply and demand predict male grooming of swollen females in captive chimpanzees, Pan troglodytes

abstract: The notion of a biological marketplace as a framework for explaining cooperative acts among animals is well supported. However, direct evidence for a biological market in a mating context is limited to a handful of studies. We collected data over a 3-year period to test supply/demand predictions of male grooming of swollen females according to a biological market in captive chimpanzees while controlling for other potential interchange. Females received more grooming from and gave less grooming to males when swollen than when not swollen, with parous females receiving more grooming from males than nulliparous females. The central tenet of a biological market that the market value of a commodity should vary according to its availability was supported. Males groomed swollen females less as the availability of swollen females in the group increased. There was also an effect of male dominance rank such that higher ranking males groomed swollen females less than lower ranking males. Taken together, our findings provide evidence for male grooming of swollen females according to a biological market while controlling for potential confounds.

title: Supply and Demand Determine the Market Value of Access to Infants in the Golden Snub-Nosed Monkey (Rhinopithecus roxellana)
abstract: According to a biological market paradigm, trading decisions between partners will be influenced by the current ‘exchange rate’ of commodities (good and services), which is affected by supply and demand, and the trader’s ability to outbid competitors. In several species of nonhuman primates, newborn infants are attractive to female group members and may become a desired commodity that can be traded for grooming within a biological market place. We investigated whether grooming was interchanged for infant handling in female golden snub-nosed monkeys (Rhinopithecus roxellana) inhabiting the Qinling Mountains of central China. R. roxellana exhibit a multilevel social organization characterized by over 100 troop members organized into 6–11 one-male units each composed one adult male and several adult females and their offspring. Behavioral data were collected over the course of 28 months on grooming patterns between mothers with infants less than 6 months old (N = 36) and other adult female troop members. Our results provide strong evidence for the interchange of grooming for access to infants. Grooming for infant access was more likely to be initiated by potential handlers (nonmothers) and less likely reciprocated by mothers. Moreover, grooming bout duration was inversely related to the number of infants per female present in each one-male unit indicating the possibility of a supply and demand market effect. The rank difference between mothers and handlers was negatively correlated with grooming duration. With increasing infant age, the duration of grooming provided by handlers was shorter suggesting that the ‘value’ of older infants had decreased. Finally, frequent grooming partners were allowed to handle and maintain access to infants longer than infrequent groomers. These results support the contention that grooming and infant handling may be traded in R. roxellana and that the price individuals paid for access to infants fluctuated with supply and demand.

title: Mutualisms in a changing world: an evolutionary perspective.
abstract: Ecology Letters (2010) 13: 1459-1474  There is growing concern that rapid environmental degradation threatens mutualistic interactions. Because mutualisms can bind species to a common fate, mutualism breakdown has the potential to expand and accelerate effects of global change on biodiversity loss and ecosystem disruption. The current focus on the ecological dynamics of mutualism under global change has skirted fundamental evolutionary issues. Here, we develop an evolutionary perspective on mutualism breakdown to complement the ecological perspective, by focusing on three processes: (1) shifts from mutualism to antagonism, (2) switches to novel partners and (3) mutualism abandonment. We then identify the evolutionary factors that may make particular classes of mutualisms especially susceptible or resistant to breakdown and discuss how communities harbouring mutualisms may be affected by these evolutionary responses. We propose a template for evolutionary research on mutualism resilience and identify conservation approaches that may help conserve targeted mutualisms in the face of environmental change.

title: Mediating mutualisms: farm management practices and evolutionary changes in symbiont co-operation
abstract: Root symbionts (rhizobia and arbuscular mycorrhizae) are often assumed to increase agricultural productivity consistently. However, rhizobial and mycorrhizal strains vary in effectiveness, resulting in symbiotic associations that range from parasitic to mutualistic. 2. The extent to which different farming practices mediate evolutionary changes along this continuum of symbiont effectiveness is rarely discussed. However, evolutionary theory suggests that (i) fertilizer use will favour parasitism unless host-plants impose sanctions against less-effective mutualists; (ii) tillage will have contrasting effects because it decreases within-plant symbiont relatedness but also decreases the risk that mutualism will benefit future competitors; (iii) crop rotation can act as a selective agent against dominating symbiont genotypes; and (iv) rhizobial inoculation adds beneficial strains to the soil but may increase the frequency of mixed nodules that allow parasitic strains to escape host sanctions. 3. However, the existing empirical data are inadequate to test our predictions thoroughly. Changes in species composition have been documented as a result of management practices, but evolutionary changes in symbiont effectiveness are difficult to detect. Therefore, a major aim of this study was to stimulate research that will assesses directly changes in symbiotic effectiveness as a function of management practices.

title: Resource stoichiometry elucidates the structure and function of arbuscular mycorrhizas across scales.
abstract: Despite the fact that arbuscular mycorrhizal (AM) associations are among the most ancient, abundant and important symbioses in terrestrial ecosystems, there are currently few unifying theories that can be used to help understand the factors that control their structure and function. This review explores how a stoichiometric perspective facilitates integration of three complementary ecological and evolutionary models of mycorrhizal structure and function. AM symbiotic function should be governed by the relative availability of carbon, nitrogen and phosphorus (trade balance model) and allocation to plant and fungal structures should depend on the availabilities of these resources (functional equilibrium model). Moreover, in an evolutionary framework, communities of plants and AM fungi are predicted to adapt to each other and their local soil environment (co-adaptation model). Anthropogenic enrichment of essential resources in the environment is known to impact AM symbioses. A more predictive theory of AM structure and function will help us to better understand how these impacts may influence plant communities and ecosystem properties.

title: The Origins of Cooperative Bacterial Communities
abstract: Bacteria live in complex multispecies communities. Intimately interacting bacterial cells are ubiquitous on biological and mineral surfaces in all habitats. Molecular and cellular biologists have unraveled some key mechanisms that modulate bacterial interactions, but the ecology and evolution of these associations remain poorly understood. One debate has focused on the relative importance of cooperation among cells in bacterial communities. Some researchers suggest that communication and cooperation, both within and among bacterial species, have produced emergent properties that give such groups a selective advantage. Evolutionary biologists have countered that the appearance of group-level traits should be viewed with caution, as natural selection almost invariably favors selfishness. A recent theory by Morris, Lenski, and Zinser, called the Black Queen Hypothesis, gives a new perspective on this debate (J. J. Morris, R. E. Lenski, and E. R. Zinser, mBio 3(2):e00036-12, 2012). These authors present a model that reshapes a decades-old idea: cooperation among species can be automatic and based upon purely selfish traits. Moreover, this hypothesis stands in contrast to the Red Queen Hypothesis, which states that species are in constant evolutionary conflict. Two assumptions serve as the core of the Black Queen model. First, bacterial functions are often leaky, such that cells unavoidably produce resources that benefit others. Second, the receivers of such by-products will tend to delete their own costly pathways for those products, thus building dependency into the interactions. Although not explicitly required in their model, an emergent prediction is that the initiation of such dependency can favor the spread of more obligate coevolved partnerships. This new paradigm suggests that bacteria might often form interdependent cooperative interactions in communities and moreover that bacterial cooperation should leave a clear genomic signature via complementary loss of shared diffusible functions.

title: The Black Queen Hypothesis: Evolution of Dependencies through Adaptive Gene Loss
abstract: Reductive genomic evolution, driven by genetic drift, is common in endosymbiotic bacteria. Genome reduction is less common in free-living organisms, but it has occurred in the numerically dominant open-ocean bacterioplankton Prochlorococcus and “Candidatus Pelagibacter,” and in these cases the reduction appears to be driven by natural selection rather than drift. Gene loss in free-living organisms may leave them dependent on cooccurring microbes for lost metabolic functions. We present the Black Queen Hypothesis (BQH), a novel theory of reductive evolution that explains how selection leads to such dependencies; its name refers to the queen of spades in the game Hearts, where the usual strategy is to avoid taking this card. Gene loss can provide a selective advantage by conserving an organism’s limiting resources, provided the gene’s function is dispensable. Many vital genetic functions are leaky, thereby unavoidably producing public goods that are available to the entire community. Such leaky functions are thus dispensable for individuals, provided they are not lost entirely from the community. The BQH predicts that the loss of a costly, leaky function is selectively favored at the individual level and will proceed until the production of public goods is just sufficient to support the equilibrium community; at that point, the benefit of any further loss would be offset by the cost. Evolution in accordance with the BQH thus generates “beneficiaries” of reduced genomic content that are dependent on leaky “helpers,” and it may explain the observed nonuniversality of prototrophy, stress resistance, and other cellular functions in the microbial world.

title: Individual-Level Bet Hedging in the Bacterium Sinorhizobium meliloti
abstract: The expression of phenotypic variability can enhance geometric mean fitness and act as a bet-hedging strategy in unpredictable environments. Metazoan bet hedging usually involves phenotypic diversification among an individual's offspring, such as differences in seed dormancy. Virtually all known microbial bet-hedging strategies, in contrast, rely on low-probability stochastic switching of a heritable phenotype by individual cells in a clonal group. This is less effective at generating within-group diversity when group size is small. Here we describe a novel microbial bet-hedging behavior that resembles individual-level metazoan bet hedging. Sinorhizobium meliloti stores carbon and energy in poly-3-hydroxybutyrate (PHB) as a contingency against carbon scarcity. We show that, when starved, dividing S. meliloti bet hedge by forming two daughter cells with different phenotypes. These have high and low PHB levels and are suited to long- and short-term starvation, respectively. The low-PHB cells have greater competitiveness for resources, whereas the high-PHB cells can survive for over a year without food, perhaps until a legume host is next available.

title: Bacterial persistence and bet hedging in Sinorhizobium meliloti.
abstract: We recently described a novel bethedging mechanism in which the bacterium Sinorhizobium meliloti responds to starvation by forming two discrete cell types via cell division. The old-pole daughter cell retains most of the resource, polyhydroxybutyrate (PHB) and is capable of surviving long-term starvation, while the low-PHB, new-pole daughter cell is capable of quickly resuming growth when starvation ends. Here we present additional data showing that the high-PHB, old-pole cells are similar to bacterial persisters, characterized by metabolic dormancy and antibiotic tolerance. Using two independent methods, we generated clonal populations of S. meliloti that varied in the frequency of the high- and low-PHB phenotypes, and then challenged these populations with ampicillin. Populations containing more high-PHB cells were significantly more antibiotic-tolerant. In a separate experiment, we used GFP fluorescence as a marker of overall metabolic activity. After 24 hours of starvation, new-pole cells were 64% brighter than their old-pole sister cells, demonstrating that the divergence in metabolic rate is rapid.

title: Bacterial farming by the fungus Morchella crassipes
abstract: The interactions between bacteria and fungi, the main actors of the soil microbiome, remain poorly studied. Here, we show that the saprotrophic and ectomycorrhizal soil fungus Morchella crassipes acts as a bacterial farmer of Pseudomonas putida, which serves as a model soil bacterium. Farming by M. crassipes consists of bacterial dispersal, bacterial rearing with fungal exudates, as well as harvesting and translocation of bacterial carbon. The different phases were confirmed experimentally using cell counting and 13C probing. Common criteria met by other non-human farming systems are also valid for M. crassipes farming, including habitual planting, cultivation and harvesting. Specific traits include delocalization of food production and consumption and separation of roles in the colony (source versus sink areas), which are also found in human agriculture. Our study evidences a hitherto unknown mutualistic association in which bacteria gain through dispersal and rearing, while the fungus gains through the harvesting of an additional carbon source and increased stress resistance of the mycelium. This type of interaction between fungi and bacteria may play a key role in soils.

title: Darwinian Agriculture: When Can Humans Find Solutions Beyond The Reach of Natural Selection?
abstract: Progress in genetic improvement of crop yield potential has slowed since 1985. Simultaneously, more sustainable management of agricultural ecosystems is needed. A better understanding of natural selection can help solve both problems. We illustrate this point with two specific examples. First, the genetic legacy of crop plants has been refined by millions of years of natural selection, often driven by competition among plants. We therefore suggest that most simple, tradeoff‐free options to increase competitiveness (e.g., increased gene expression, or minor modifications of existing plant genes) have already been tested by natural selection. Further genetic improvement of crop yield potential over the next decade will mainly involve tradeoffs, either between fitness in past versus present environments, or between individual competitiveness and the collective performance of plant communities. Eventually, we may develop the ability to predict the consequences of genetic alterations so radical that they have not yet been tested by natural selection. Second, natural selection acts mainly at the level of genes, individuals, and family groups, rather than ecosystems as a whole. Consequently, there is no reason to expect the structure of natural ecosystems (diversity, spatial, or temporal patterns) to be a reliable blueprint for agricultural ecosystems. Natural ecosystems are nonetheless an important source of information that could be used to improve agriculture.

title: The Evolution of Mutualism in Gut Microbiota Via Host Epithelial Selection
abstract: A computer model of the gut shows how a host can readily select friendly bacteria over harmful bacteria through a process called “selectivity amplification.”

title: Host-Bacterial Mutualism in the Human Intestine
abstract: The distal human intestine represents an anaerobic bioreactor programmed with an enormous population of bacteria, dominated by relatively few divisions that are highly diverse at the strain/subspecies level. This microbiota and its collective genomes (microbiome) provide us with genetic and metabolic attributes we have not been required to evolve on our own, including the ability to harvest otherwise inaccessible nutrients. New studies are revealing how the gut microbiota has coevolved with us and how it manipulates and complements our biology in ways that are mutually beneficial. We are also starting to understand how certain keystone members of the microbiota operate to maintain the stability and functional adaptability of this microbial organ.

title: Animal Behavior and the Microbiome
abstract: Feedbacks between microbiomes and their hosts affect a range of animal behaviors. Human bodies house trillions of symbiotic microorganisms. The genes in this human microbiome outnumber human genes by 100 to 1, and their study is providing profound insights into human health. But humans are not the only animals with microbiomes, and microbiomes do not just impact health. Recent research is revealing surprising roles for microbiomes in shaping behaviors across many animal taxa—shedding light on how behaviors from diet to social interactions affect the composition of host-associated microbial communities (1, 2), and how microbes in turn influence host behavior in dramatic ways (2–6).