Characterisation of rhizobia for the new annual pasture legume Scorpiurus muricatus targeted for medium-to-low rainfall areas of southern Australia

Legumes play an integral role in increasing agricultural productivity, particularly in low input agricultural systems in Australia, due to their ability to form symbiotic interactions with a group of soil bacteria called rhizobia. However, in medium-to-low rainfall areas of southern Australia, there is a lack of suitable annual pasture legumes, which is limiting agricultural productivity and profitability in these farming systems. Scorpiurus muricatus is an annual legume from the Mediterranean which possesses high nutritive value and palatability for livestock, is high yielding, capable of self-seeding and is well-adapted to hot and dry summers. As such, S. muricatus is currently being evaluated as a new pasture legume for southern Australia. Crucial to the success of introducing this legume will be the availability of a highly effective rhizobial inoculant strain. This thesis therefore sought to characterise the phylogeny, free-living and symbiotic phenotype of a range of bacteria isolated from Scorpiurus spp. A total of 19 strains were investigated, with 16s rRNA sequencing demonstrating that 18 of these strains belonged to the genus Mesorhizobium, with the remaining strain (WSM1184) most closely related to Agrobacterium tumefaciens. Analysis of nifH and nodC symbiosis genes further showed that the characterised Mesorhizobium strains generally shared highly similar sequences for these loci, indicating a comparatively high degree of genetic similarity. In particular, WSM1343 (isolated from Scorpiurus sulcatus growing in Morocco) and WSM1386 (isolated from S. sulcatus in Manjimup, Western Australia) were shown to share highly similar symbiosis genes, but divergent 16S rRNA genes, suggesting the possibility that these strains may contain symbiosis genes on mobile Integrative and Conjugative Elements (ICEs). While the temperature tolerance and apparent optimum growth temperature of the test strains of 28°C was consistent with that commonly reported for Mesorhizobium s