Kin discrimination drives territorial exclusion during Bacillus subtilis swarming and restrains exploitation of surfactin

Swarming is the collective movement of bacteria across a surface. It requires the production of surfactants (public goods) to overcome surface tension and provides an excellent model to investigate bacterial cooperation. Previously, we correlated swarm interaction phenotypes with kin discrimination between B. subtilis soil isolates, by showing that less related strains form boundaries between swarms and highly related strains merge. However, how kin discrimination affects cooperation and territoriality in swarming bacteria remains little explored. Here we show that the pattern of surface colonization by swarming mixtures is influenced by kin types. Closely related strain mixtures colonize the surface in a mixed swarm, while mixtures of less related strains show competitive exclusion as only one strain colonizes the surface. The outcome of nonkin swarm expansion depends on the initial ratio of the competing strains, indicating positive frequency-dependent competition. We find that addition of surfactin (a public good excreted from cells) can complement the swarming defect of nonkin mutants, whereas close encounters in nonkin mixtures lead to territorial exclusion, which limits the exploitation of surfactin by nonkin nonproducers. The work suggests that kin discrimination driven competitive territorial exclusion may be an important determinant for the success of cooperative surface colonization.

Kin discrimination modifies fitness, spatial segregation and matrix sharing between strains with low relatedness in Bacillus subtilis biofilms

Microorganisms in nature form multicellular groups called biofilms. In biofilms bacteria embedded in a matrix of extracellular polymeric substances (EPS) interact intensely, due to their proximity to each other. Most studies have investigated genetically homogeneous biofilms, leaving a gap in knowledge on genetically heterogeneous biofilms. Recent insights show that a Gram-positive model bacterium, Bacillus subtilis, discriminates between strains of high (kin) and low (non-kin) phylogenetic relatedness, reflected in merging (kin) and boundaries (non-kin) between swarms. However, it is not clear how kinship between interacting strains affects their fitness, the genotype distribution, and the EPS sharing in floating biofilms (pellicles). To address this gap in knowledge we cultivate B. subtilis strains as mixtures of kin and non-kin strains in static cultures, allowing them to form pellicles. We show here that in non-kin pellicles only one strain’s fitness was reduced; at the same time, strains segregated into larger patches and exhibited decreased matrix sharing, as compared to kin and isogenic pellicles, in which both strains had comparable colony forming units (CFU) counts and more homogenous cell mixing. Overall, our results emphasize kin discrimination (KD) as a social behavior that shapes fitness, spatial segregation and sharing of the extracellular matrix in genetically heterogenous biofilms of B. subtilis.

Family-specific chemical profiles provide potential kin recognition cues in the sexually cannibalistic spider Argiope bruennichi

Kin recognition, the ability to detect relatives, is important for cooperation, altruism and also inbreeding avoidance. A large body of research on kin recognition mechanisms exists for vertebrates and insects, while little is known for other arthropod taxa. In spiders, nepotism has been reported in social and solitary species. However, there are very few examples of kin discrimination in a mating context, one coming from the orb-weaver Argiope bruennichi . Owing to effective mating plugs and high rates of sexual cannibalism, both sexes of A. bruennichi are limited to a maximum of two copulations. Males surviving their first copulation can either re-mate with the current female (monopolizing paternity) or leave and search for another. Mating experiments have shown that males readily mate with sisters but are more likely to leave after one short copulation as compared with unrelated females, allowing them to search for another mate. Here, we ask whether the observed behaviour is based on chemical cues. We detected family-specific cuticular profiles that qualify as kin recognition cues. Moreover, correlations in the relative amounts of some of the detected substances between sexes within families indicate that kin recognition is likely based on subsets of cuticular substances, rather than entire profiles.

Kin discrimination in allelopathy and consequences for agriculture

Recent research has shown that plants can distinguish genetically-related individuals from strangers (kin recognition) and exhibit more cooperative behaviours towards these more related individuals (kin discrimination). The first evidence for this was found when Cakile edentula plants growing with half-sibs allocated relatively less biomass to roots than plants growing with unrelated individuals, indicating that kin recognition can reduce the intensity of competition (Dudley & File, 2007). Since then, kin discrimination has been shown to result in reduced competition for soil resources (Semchenko, Saar, & Lepik, 2014), light (Crepy & Casal, 2015) and pollinators (Torices, Gómez, & Pannell, 2018). On the other hand, allelopathy, plants producing chemical compounds that negatively affect performance of neighbour plants, has also been widely documented (Inderjit & Duke, 2003) and shown to profoundly affect local species coexistence and plant community structure (Meiners, Kong, Ladwig, Pisula, & Lang, 2012). In crops allelopathy can also be beneficial in suppressing weeds (Macías, Mejías, & Molinillo, 2019). In the current issue, Xu, Cheng, Kong, and Meiners (2021) published the first study to show that kin discrimination can also affect the balance between direct competition for resources and allelopathy, and this together may lead to improved weed suppression in rice.

Kin discrimination promotes horizontal gene transfer between unrelated strains in Bacillus subtilis

Bacillus subtilis is a soil bacterium that is competent for natural transformation. Genetically distinct B. subtilis swarms form a boundary upon encounter, resulting in killing of one of the strains. This process is mediated by a fast-evolving kin discrimination (KD) system consisting of cellular attack and defence mechanisms. Here, we show that these swarm antagonisms promote transformation-mediated horizontal gene transfer between strains of low relatedness. Gene transfer between interacting non-kin strains is largely unidirectional, from killed cells of the donor strain to surviving cells of the recipient strain. It is associated with activation of a stress response mediated by sigma factor SigW in the donor cells, and induction of competence in the recipient strain. More closely related strains, which in theory would experience more efficient recombination due to increased sequence homology, do not upregulate transformation upon encounter. This result indicates that social interactions can override mechanistic barriers to horizontal gene transfer. We hypothesize that KD-mediated competence in response to the encounter of distinct neighbouring strains could maximize the probability of efficient incorporation of novel alleles and genes that have proved to function in a genomically and ecologically similar context.

Size-dependent tradeoffs in aggressive behavior towards kin

Aggression between juveniles can be unexpected, as their primary motivation is to survive until their reproductive stage. However, instances of aggression, which may escalate to cannibalism, can be vital for survival, although the factors (e.g. genetic or environmental) leading to cannibalism vary across taxa. While cannibalism can greatly accelerate individual growth, it may also reduce inclusive fitness when kin are consumed. As a solution to this problem, some cannibals demonstrate kin discrimination and preferentially attack unrelated individuals. Here, we used both experimental and modeling approaches to consider how physical traits (e.g. size in relation to opponent) and genetic relatedness mediate aggressive behavior in dyads of cannibalistic Dendrobates tinctorius tadpoles. We paired sibling, half-sibling, and non-sibling tadpoles of different sizes together in an arena and recorded their aggression and activity. We found that the interaction between size and relatedness predicts aggressive behavior: large non-siblings are significantly more aggressive than large siblings. Unexpectedly, although siblings tended to attack less overall, in size mismatched pairs they attacked faster than in non-sibling treatments. Ultimately, it appears that larval aggression reflects a balance between relatedness and size where individuals trade-off their own fitness with that of their relatives.