1887

Abstract

STM815 is a β-rhizobial strain that can effectively nodulate several species of the large legume genus . Two Tn-induced mutants of this strain, KM16-22 and KM51, failed to form root nodules on , but still caused root hair deformation, which is one of the early steps of rhizobial infection. Both mutants grew well in a complex medium. However, KM16-22 could not grow on minimal medium unless a sugar and a metabolic intermediate such as pyruvate were provided, and KM51 also could not grow on minimal medium unless a sugar was added. The Tn-interrupted genes of the mutants showed strong homologies to , which encodes 2,3-biphosphoglycerate-dependent phosphoglycerate mutase (dPGM), and , which encodes fructose 1,6-bisphosphatase (FBPase). Both enzymes are known to be involved in obligate steps in carbohydrate metabolism. Enzyme assays confirmed that KM16-22 and KM51 had indeed lost dPGM and FBPase activity, respectively, whilst the activities of these enzymes were expressed normally in both free-living bacteria and symbiotic bacteroids of the parental strain STM815. Both mutants recovered their enzyme activity after the introduction of wild-type or genes, were subsequently able to use carbohydrate as a carbon source, and were able to form root nodules on and to fix nitrogen as efficiently as the parental strain. We conclude that the enzymes dPGM and FBPase are essential for the formation of a symbiosis with the host plant.

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2012-04-01
2019-12-05
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