Pyruvate dehydrogenase activity is important for colonization of seeds and roots by Enterobacter cloacae

Enterobacter cloacae is a plant-beneficial bacterium that shows promise for suppression of damping-off of cucumber and other crops caused by Pythium ultimum. We have been using a mutational approach to determine the E. cloacae genes important in bacterial–plant and bacterial–pathogen interactions in the spermosphere and rhizosphere. E. cloacae M43 is a transposon mutant of E. cloacae 501R3 that was significantly impaired in colonization of seeds and roots of diverse crop plants. Strain M43 did not increase in population on cucumber, sunflower, and wheat seeds and was significantly reduced in growth on pea seeds relative to strain 501R3. Populations of M43 were also dramatically lower than those of strain 501R3 in cucumber, pea, sunflower, and wheat rhizosphere in 42 d experiments. Molecular characterization of M43 demonstrated that there was a single transposon insertion in the genome of this strain and that this insertion was in a region of the E. cloacae genome with a high degree of DNA sequence identity with aceF. aceF encodes the dihydrolipoamide acetyltransferase subunit of the pyruvate dehydrogenase complex (PDHC). Cell lysates from strain 501R3 grown on minimal medium plus 50 mM glycerol and 2 mM acetate contained 0.011±0.0036 U pyruvate dehydrogenase activity while cell lysates from M43 grown under identical conditions contained no detectable pyruvate dehydrogenase activity. Additionally, the nutritional use profile of M43 under aerobic and anaerobic conditions was as expected for an ace mutant. Experiments reported here strongly suggest a role for aceF and the PDHC in colonization of seeds and roots of diverse crop plants by E. cloacae.