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Here we give names to three new species of Paraburkholderia that can remain in symbiosis indefinitely in the spores of a soil dwelling eukaryote, Dictyostelium discoideum . The new species P. agricolaris sp. nov. , P. hayleyella sp. nov. , and P. bonniea sp. nov . are widespread across the eastern USA and were isolated as internal symbionts of wild-collected D. discoideum . We describe these sp. nov. using several approaches. Evidence that they are each a distinct new species comes from their phylogenetic position, average nucleotide identity, genome-genome distance, carbon usage, reduced length, cooler optimal growth temperature, metabolic tests, and their previously described ability to invade D. discoideum amoebae and form a symbiotic relationship . All three of these new species facilitate the prolonged carriage of food bacteria by D. discoideum, though they themselves are not food. Further studies of the interactions of these three new species with D. discoideum should be fruitful for understanding the ecology and evolution of symbioses. 

A species can benefit or be hurt by other species. For example, honeybees and flowering plants help each other to flourish, while lions and gazelles behave in ways that decrease each other’s populations. Understanding these relationships is important for controlling pests and diseases. Sometimes it is easiest to study such interactions by looking at simple ones that happen on a small scale. Amoebas are common soil organisms that have the same basic organization as human cells. They are much larger and more complex than the bacteria that also live in the soil. How exactly the amoebas and bacteria interact in the soil is an important question, particularly as some of the bacteria can also live inside amoebas. Does this intimate relationship help or harm the amoeba? Shu, Brock, Geist et al. studied the relationship between a widely studied species of social amoeba and two species of bacteria that can live inside it. Some of the amoebas naturally contained one of the bacteria species, and others were infected with the bacteria in experiments. Throughout the entire life cycle of the amoebas, the bacteria lived inside them. During one part of the life cycle, amoebas form so-called fruiting bodies, which release spores that can develop into new amoebas. Shu et al. found that both types of bacteria alter the structure of the fruiting bodies in ways that reduce how well the spores disperse. One of the bacteria species, called Burkholderia hayleyella, harmed the amoebas a lot. It caused most harm to amoebas that do not naturally host the bacteria. This indicates that the amoebas that do host this species may have evolved to avoid its worst effects. The amoebas have many similarities to the white blood cells that clear bacteria from the human body. Certain bacteria can get inside white blood cells, causing diseases such as tuberculosis. Understanding how bacteria harm amoebas might be useful for understanding such diseases, and developing treatments for them. Though the bacteria Shu et al. studied are not toxic to humans, they are closely related to bacteria that are harmful. It is therefore possible that some bacteria that infect humans first evolve to infect amoebas. 

Abstract The establishment of symbioses between eukaryotic hosts and bacterial symbionts in nature is a dynamic process. The formation of such relationships depends on the life history of both partners. Bacterial symbionts of amoebae may have unique evolutionary trajectories to the symbiont lifestyle, because bacteria are typically ingested as prey. To persist after ingestion, bacteria must first survive phagocytosis. In the social amoebaDictyostelium discoideum, certain strains ofBurkholderiabacteria are able to resist amoebal digestion and maintain a persistent relationship that includes carriage throughout the amoeba's social cycle that culminates in spore formation. SomeBurkholderiastrains allow their host to carry other bacteria, as food. This carried food is released in new environments in a trait called farming. To better understand the diversity and prevalence ofBurkholderiasymbionts and the traits they impart to their amoebae hosts, we first screened 700 natural isolates ofD. discoideumand found 25% infected withBurkholderia. We next used a multilocus phylogenetic analysis and identified two independent transitions byBurkholderiato the symbiotic lifestyle. Finally, we tested the ability of 38 strains ofBurkholderiafromD. discoideum, as well as strains isolated from other sources, for traits relevant to symbiosis inD. discoideum. OnlyD. discoideumnative isolates belonging to theBurkholderia agricolaris,B. hayleyella, andB. bonnieaspecies were able to form persistent symbiotic associations withD. discoideum.TheBurkholderia–Dictyosteliumrelationship provides a promising arena for further studies of the pathway to symbiosis in a unique system.