Roles for Azorhizobial Nod Factors and Surface Polysaccharides in Intercellular Invasion and Nodule Penetration, Respectively

In the symbiotic interaction between Azorhizobium caulinodans and Sesbania rostrata root and stem-borne nodules are formed. The bacteria enter the host via intercellular spaces at lateral or adventitious root bases and form infection pockets in outer cortical layers. Infection threads guide the bacteria to nodule primordia where plant cells are invaded. To identify bacterial functions that are required for this infection process, two mutants defective in nodulation were studied; one produced no Nod factors (nodA mutant), the other had altered surface polysaccharides (SPS) and induced the formation of pseudo-nodules, Bacteria were visualized with the help of a nodA-uidA reporter fusion that was functional during nodule development and in bacteroids. In contrast to the SPS mutant, nodA mutants were unable to colonize outer cortical regions. In mixed inoculations with both mutants, functional nodules were formed, the central tissue of which was occupied by the nodA mutant. These observations suggest that SPS play a role in deeper invasion and that Nod factors are necessary for entry. Simultaneous application of purified Nod factors and nodA mutant bacteria restored the formation of outer cortical infection pockets leading to the conclusion that intercellular infection is an active process that is dependent on bacterial Nod factor signaling.