DNA homology within the Rhizobiaceae : a thesis presented in partial fulfilment of the requirements for the degree of Master of Science in Microbiology at Massey University

Abstract

The relationship of rhizobia that nodulate Galega officinalis to the known species of Rhizobium and Bradyrhizobium was investigated. Similarly, the recently discovered fast­ growing soybean nodulating group of rhizobia was studied. Both groups were investigated using DNA:DNA hybridization as well as nodulation on legumes and phage-typing.

The Galega nodulating rhizobia were found to form a distinct DNA homology group. The mean relative homology of 11 strains of Galega nodulating rhizobia with the reference strains gal 1 and gal NW 3, which effectively nodulate Galega officinalis, was significantly higher than the mean relative homology of other groups of rhizobia.

The Galega rhizobia only nodulated Galega officinalis and formed a distinct phage-typing group in agreement with the DNA homology results. These rhizobia therefore appear to form a unique taxonomic group within the genus Rhizobium.

The fast-growing soybean nodulating rhizobia formed a distinct DNA homology group with at least two subgroups. The mean relative homology of 11 of these strains with the reference strains USDA 208 and USDA 191 which nodulate Glycine max, was significantly higher than the mean relative homology of other groups of rhizobia. Low DNA homologies were found between the fast-growing soybean strains and Bradyrhizobium japonicum ATCC 10324.

The fast-growing soybean nodulating rhizobia nodulated glycine max and formed ineffective nodules on Lotus pedunculatus. None of these strains were lysed by the bacteriophages used in the study, but as yet, no bacteriophage specific for this group of rhizobia has been isolated. The fast­ growing soybean nodulating rhizobia were concluded to be taxonomically distinct from other species of Rhizobium. The thermal stability of reassociated DNA duplexes was examined for both the Galega and fast-growing soybean rhizobia and further indicated the uniqueness of both groups.

The use of colony hybridization as a means of identifying different strains of Rhizoiium was investigated and was found to be useful in distinguishing between genetically distinct rhizobia and to identify rhizobia within root nodules.