Forty rhizobia nodulating four Acacia species (A. gummifera, A. raddiana, A. cyanophylla, and A. horrida) were isolated from different sites in Morocco. These rhizobia were compared by analyzing both the 16S rRNA gene (rDNA) and the 16S-23S rRNA spacer by PCR with restriction fragment length polymorphism (RFLP) analysis. Analysis of the length of 16S-23S spacer showed a considerable diversity within these microsymbionts, but RFLP analysis of the amplified spacer revealed no additional heterogeneity. Three clusters were identified when 16S rDNA analysis was carried out. Two of these clusters include some isolates which nodulate, nonspecifically, the four Acacia species. These clusters, A and B, fit within the Sinorhizobiumlineage and are closely related to S. meliloti and S. fredii, respectively. The third cluster appeared to belong to theAgrobacterium-Rhizobium galegae phylum and is more closely related to the Agrobacterium tumefaciens species. These relations were confirmed by sequencing a representative strain from each cluster.
The phenotypic characteristics of 48 isolates obtained from root nodules of four Acacia species (Acacia cyanophylla, A. gummifera, A. horrida and A. raddiana) growing in soils collected from the arid and Saharan regions of Morocco were studied. The rhizobia were very diverse with respect to their cross‐nodulation patterns, as well as their physiological and biochemical properties. Dendrograms obtained through computer numerical analysis of 52 phenotypic characteristics showed that isolates could fit into four clusters below the boundary level of 0·85 average distance and that they were very distinct from the reference strains. Some interesting isolates for inoculation trials have been identified. They were able to grow at pH ranging from 4 to 9, tolerate a high salt concentration (3% NaCl) and grew at a maximum temperature between 35 and 40 °C.
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