Mutualistic co‐evolution of T3SSs during the establishment of symbiotic relationships between Vigna radiata and Bradyrhizobia

This study supports the idea that the evolution of type III secretion system (T3SS) is one of the factors that controls Vigna radiata–bradyrhizobia symbiosis. Based on phylogenetic tree data and gene arrangements, it seems that the T3SSs of the Thai bradyrhizobial strains SUTN9‐2, DOA1, and DOA9 and the Senegalese strain ORS3257 may share the same origin. Therefore, strains SUTN9‐2, DOA1, DOA9, and ORS3257 may have evolved their T3SSs independently from other bradyrhizobia, depending on biological and/or geological events. For functional analyses, the rhcJ genes of ORS3257, SUTN9‐2, DOA9, and USDA110 were disrupted. These mutations had cultivar‐specific effects on nodulation properties. The T3SSs of ORS3257 and DOA9 showed negative effects on V. radiata nodulation, while the T3SS of SUTN9‐2 showed no effect on V. radiata symbiosis. In the roots of V. radiata CN72, the expression levels of the PR1 gene after inoculation with ORS3257 and DOA9 were significantly higher than those after inoculation with ORS3257 ΩT3SS, DOA9 ΩT3SS, and SUTN9‐2. The T3Es from ORS3257 and DOA9 could trigger PR1 expression, which ultimately leads to abort nodulation. In contrast, the T3E from SUTN9‐2 reduced PR1 expression. It seems that the mutualistic relationship between SUTN9‐2 and V. radiata may have led to the selection of the most well‐adapted combination of T3SS and symbiotic bradyrhizobial genotype.