Azospirillum–Rhizobium interaction leading to a plant growth stimulation without nodule formation

Abstract

The effect of inoculation of white clover plants with mixed cultures of Rhizobium trifolii strain ANU870 and Azospirillum brasilense strain SP245 was examined. Ratios of Rhizobium–Azospirillum (R:A) of 1:200 to 1:2500 caused an inhibition of nodulation. However, these nonnodulated plants did not show nitrogen-deficiency symptoms when grown on nitrogenfree medium. When these plants were assayed for acetylene reduction activity a low level of ethylene production was detected. A significant increase in plant dry weights was also observed. Isolation of viable bacterial cells from surface-sterilized root segments of plants inoculated with an R:A ratio of 1:200 revealed that 80% of the bacterial population was made up of the Azospirillum strain. Under laboratory conditions transfer of the Rhizobium Sym(biosis) plasmid pBRIAN to strain SP245 was observed ex planta. However, the Sym plasmid was unstable in Azospirillum. A high frequency of Tn5 transfer from pBRIAN to strain SP245 occurred when strains ANU870 and SP245 were mixed in the rhizosphere and (or) in the root tissue. Tn5 transposed preferentially into the smallest indigenous plasmid of strain SP245 and was easily lost when this strain (SP245::Tn5) was not maintained on selective medium. This mutated Azospirillum strain caused plant growth stimulation when inoculated onto white clover plants.