Genetic organization and functional analysis of the type III secretion system of Bradyrhizobium elkanii.

Cloning and sequencing of a 47.1-kb chromosomal DNA region revealed the presence of a type III secretion system (T3SS) in Bradyrhizobium elkanii USDA61. The identified genes are likely to encode the transcriptional activator TtsI, core components of the secretion apparatus and secreted proteins. Several ORFs within the cluster are not conserved in other rhizobia. Nine tts box motifs, a promoter element of TtsI-regulated genes, were found; six of them upstream of annotated genes. For functional analyses, the rhcC2 and rhcJ genes were disrupted. These mutations had a cultivar-specific effect on nodulation. Vigna radiata cv. KPS1 developed nodules if infected with the mutant strains but not with the wild type. In contrast, V. radiata cv. CN36 was nodulated by all strains. Nodulation of rj(1) soybean depended on the T3SS. A comparison of the protein patterns from supernatants of the wild type and rhcJ mutant by two-dimensional gel electrophoresis revealed proteins that are secreted only in the wild-type background. These results show that B. elkanii encodes a functional T3SS that is involved in the interaction with host legumes.