title: Experimental evolution of nodule intracellular infection in legume symbionts.
abstract: Soil bacteria known as rhizobia are able to establish an endosymbiosis with legumes that takes place in neoformed nodules in which intracellularly hosted bacteria fix nitrogen. Intracellular accommodation that facilitates nutrient exchange between the two partners and protects bacteria from plant defense reactions has been a major evolutionary step towards mutualism. Yet the forces that drove the selection of the late event of intracellular infection during rhizobium evolution are unknown. To address this question, we took advantage of the previous conversion of the plant pathogen Ralstonia solanacearum into a legume-nodulating bacterium that infected nodules only extracellularly. We experimentally evolved this draft rhizobium into intracellular endosymbionts using serial cycles of legume-bacterium cocultures. The three derived lineages rapidly gained intracellular infection capacity, revealing that the legume is a highly selective environment for the evolution of this trait. From genome resequencing, we identified in each lineage a mutation responsible for the extracellular-intracellular transition. All three mutations target virulence regulators, strongly suggesting that several virulence-associated functions interfere with intracellular infection. We provide evidence that the adaptive mutations were selected for their positive effect on nodulation. Moreover, we showed that inactivation of the type three secretion system of R. solanacearum that initially allowed the ancestral draft rhizobium to nodulate, was also required to permit intracellular infection, suggesting a similar checkpoint for bacterial invasion at the early nodulation/root infection and late nodule cell entry levels. We discuss our findings with respect to the spread and maintenance of intracellular infection in rhizobial lineages during evolutionary times.
authors: INRA, Laboratoire des Interactions Plantes-Microorganismes (LIPM), UMR441, Castanet-Tolosan, France.
authors: Su Hua
authors: SH
authors: Guan
authors: Carine
authors: C
authors: Gris
authors: Stéphane
authors: S
authors: Cruveiller
authors: Cécile
authors: C
authors: Pouzet
authors: Lena
authors: L
authors: Tasse
authors: Aurélie
authors: A
authors: Leru
authors: Aline
authors: A
authors: Maillard
authors: Claudine
authors: C
authors: Médigue
authors: Jacques
authors: J
authors: Batut
authors: Catherine
authors: C
authors: Masson-Boivin
authors: Delphine
authors: D
authors: Capela
mesh_terms: D058947:Bacterial Secretion Systems / Q000235:genetics; D005075:Biological Evolution; D007887:Fabaceae / Q000382:microbiology*; D009154:Mutation; D018517:Plant Roots / Q000382:microbiology; D012231:Rhizobium / Q000235:genetics* / Q000472:pathogenicity; D013559:Symbiosis / Q000235:genetics*; D037521:Virulence Factors / Q000235:genetics
chemical_list: D058947:Bacterial Secretion Systems; D037521:Virulence Factors