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Riemerella anatipestifer  is a gram-negative bacterium that is the first naturally competent bacterium identified in the family  Flavobacteriaceae . However, the determinants that influence the natural transformation and the underlying mechanism remain unknown. In this study, we evaluated the effects of various nutritional factors of the GCB medium [glucose, L-glutamine, vitamin B1, Fe (NO 3 ) 3 , NaCl, phosphate, and peptone], on the natural transformation of  R. anatipestifer  ATCC 11845. Among the assayed nutrients, peptone and phosphate affected the natural transformation of  R. anatipestifer  ATCC 11845, and the transformation frequency was significantly decreased when phosphate or peptone was removed from the GCB medium. When the iron chelator 2,2′-dipyridyl (Dip) was added, the transformation frequency was decreased by approximately 100-fold and restored gradually when Fe (NO 3 ) 3  was added, suggesting that the natural transformation of  R. anatipestifer  ATCC 11845 requires iron. Given the importance of TonB in nutrient transportation, we further identified whether TonB is involved in the natural transformation of  R. anatipestifer  ATCC 11845. Mutation of  tonBA  or  tonBB , but not  tbfA , was shown to inhibit the natural transformation of  R. anatipestifer  ATCC 11845 in the GCB medium. In parallel, it was shown that the  tonBB  mutant, but not the  tonBA  mutant, decreased iron acquisition in the GCB medium. This result suggested that the  tonBB  mutant affects the natural transformation frequency due to the deficiency of iron utilization.

Effect of Nutritional Determinants and TonB on the Natural Transformation of Riemerella anatipestifer