Multilocus sequence analysis of root nodule isolates from Lotus arabicus (Senegal), Lotus creticus, Argyrolobium uniflorum and Medicago sativa (Tunisia) and description of Ensifer numidicus sp. nov. and Ensifer garamanticus sp. nov.

Nine isolates from Argyrolobium uniflorum, Lotus creticus , Medicago sativa (Tunisia) and Lotus arabicus (Senegal) were analysed by multilocus sequence analysis (MLSA) of five housekeeping genes (recA, atpD, glnA, gltA and thrC), the 16S rRNA gene and the nodulation gene nodA. Analysis of the individual and concatenated gene sequences demonstrated that the nine new strains constituted three stable, well-supported (bootstrap and gene sequence similarity values) monophyletic clusters, A, B and C, all belonging to the branch of the genus Ensifer, regardless of the phylogenetic reconstruction method used (maximum likelihood, maximum-parsimony, neighbour-joining). The three groups were further characterized by API 100 auxanographic tests, host specificity and nodA gene sequence analysis. On the basis of these data, clusters A and C are suggested as representing two novel species within the genus Ensifer, for which the names Ensifer numidicus sp. nov. (type strain ORS 1407(T)=LMG 24690(T)=CIP 109850(T)) and Ensifer garamanticus sp. nov. (type strain ORS 1400(T)=LMG 24692(T)=CIP 109916(T)) are proposed. The cluster B strains were assigned to Ensifer adhaerens genomovar A.

Phenotypical and genotypical characteristics of root-nodulating bacteria isolated from annual Medicago spp. in Soummam Valley (Algeria).

AIMS: In the framework of agro-pastoral system management using local annual medics coupled with their native root-nodulating bacteria to extend pasture zones, increase forage yields and improve ovine and bovine breeding in Algeria, we investigated diversity of rhizobia from annual Medicago spp. (Medicago arabica, Medicago polymorpha, Medicago minima and Medicago orbicularis). METHODS AND RESULTS: Ten nodulating-isolates were characterized by morphological, cultural, physiological and biochemical features, SDS-PAGE analysis and PCR-RFLP of 16S rDNA. The results show some degree of genetic diversity among the isolates; three can be affiliated to Sinorhizobium meliloti, one to Rhizobium galegae and six were separate. CONCLUSIONS: Local annual medics would have a high degree of specificity in their symbiotic interaction. Furthermore, our results support the presence of Rh. galegae in the Mediterranean region. SIGNIFICANCE AND IMPACT OF THE STUDY: This work is a preliminary step towards selection of efficient symbiotic Medicago-rhizobia to develop inoculants for management of agro-pastoral systems using local annual medics in Algeria.