Characterization of the indole-3 acetic acid (IAA) biosynthetic pathway in an epiphytic strain of Erwinia herbicola and IAA production in vitro

An epiphytic strain of Erwinia herbicola (strain 299R) synthesized indole-3-acetic acid (IAA) from indole-3-pyruvic acid and indole-3-acetaldehyde, but not from indole-3-acetamide and other intermediates of various IAA biosynthetic pathways in enzyme assays. TLC, HPLC, and CC-MS analyses revealed the presence of indole-3-pyruvic acid, indole-3-ethanol, and IAA in culture supernatants of strain 299R. Indole-3-acetaldehyde was detected in enzyme assays. Furthermore, strain 299R genomic DNA shared no homology with the iaaM and iaaH genes from Pseudomonas syringae pv. savastanoi, even in Southern hybridizations performed under low-stringency conditions. These observations strongly suggest that unlike gall-forming bacteria which can synthesize IAA by indole-3-acetamide, the indole-3-pyruvic acid pathway is the primary route for IAA biosynthesis in this plant-associated strain. IAA synthesis in tryptophan-supplemented cultures of strain 299R was over 10-fold higher under nitrogen-limiting conditions, indicating a possible role for IAA production by bacterial epiphytes in the acquisition of nutrients during growth in their natural habitat.