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1. The Bradyrhizobium Sp. LmicA16 Type VI Secretion System Is Required for Efficient Nodulation of Lupinus Spp.

Author

De Sousa, B. F. S., Ruiz-Argueso, T., Tighilt, L, Palacios, J. M, Imperial, J., Rey, L., Boulila, F, Giraud, E., De Sousa, B. F. S., Ruiz-Argueso, T., Tighilt, L, Palacios, J. M, Imperial, J., Rey, L., Boulila, F, and Giraud, E.

Abstract

Many bacteria of the genus Bradyrhizobium are capable of inducing nodules in legumes. In this work, the importance of a type VI secretion system (T6SS) in a symbiotic strain of the genus Bradyrhizobium is described. T6SS of Bradyrhizobium sp. LmicA16 (A16) is necessary for efficient nodulation with Lupinus micranthus and Lupinus angustifolius. A mutant in the gene vgrG, coding for a component of the T6SS nanostructure, induced less nodules and smaller plants than the wild-type (wt) strain and was less competitive when co-inoculated with the wt strain. A16 T6SS genes are organized in a 26-kb DNA region in two divergent gene clusters of nine genes each. One of these genes codes for a protein (Tsb1) of unknown function but containing a methyltransferase domain. A tsb1 mutant showed an intermediate symbiotic phenotype regarding vgrG mutant and higher mucoidity than the wt strain in free-living conditions. T6SS promoter fusions to the lacZ reporter indicate expression in nodules but not in free-living cells grown in different media and conditions. The analysis of nodule structure revealed that the level of nodule colonization was significantly reduced in the mutants with respect to the wt strain.

Published
2021

2. Cytisus villosus from Northeastern Algeria is nodulated by genetically diverse Bradyrhizobium strains.

Author

Ahnia H, Boulila F, Boulila A, Boucheffa K, Durán D, Bourebaba Y, Salmi A, Imperial J, Ruiz-Argüeso T, and Rey L

Subjects
Algeria, Bacterial Proteins genetics, Bradyrhizobium genetics, Cluster Analysis, DNA, Bacterial chemistry, DNA, Bacterial genetics, DNA, Ribosomal chemistry, DNA, Ribosomal genetics, Genes, Bacterial, Lupinus microbiology, Lupinus physiology, Molecular Sequence Data, Multilocus Sequence Typing, Phylogeny, Polymerase Chain Reaction, RNA, Ribosomal, 16S genetics, Random Amplified Polymorphic DNA Technique, Spartium microbiology, Spartium physiology, Bradyrhizobium classification, Bradyrhizobium isolation & purification, Cytisus microbiology, Cytisus physiology, Plant Root Nodulation
Abstract

Fifty-one rhizobial strains isolated from root nodules of Cytisus villosus growing in Northeastern Algeria were characterized by genomic and phenotypic analyses. Isolates were grouped into sixteen different patterns by PCR-RAPD. The phylogenetic status of one representative isolate from each pattern was examined by multilocus sequence analyses of four housekeeping genes (16S rRNA, glnII, recA, and atpD) and one symbiotic gene (nodC). Analysis of 16S rRNA gene sequences showed that all the isolates belonged to the genus Bradyrhizobium. Phylogenetic analyses based on individual or concatenated genes glnII, recA, and atpD indicated that strains cluster in three distinct groups. Ten out of the sixteen strains grouped together with Bradyrhizobium japonicum, while a second group of four clustered with Bradyrhizobium canariense. The third group, represented by isolates CTS8 and CTS57, differed significantly from all other bradyrhizobia known to nodulate members of the Genisteae tribe. In contrast with core genes, sequences of the nodC symbiotic gene from all the examined strains form a homogeneous group within the genistearum symbiovar of Bradyrhizobium. All strains tested nodulated Lupinus angustifolius, Lupinus luteus, and Spartium junceum but not Glycine max. From these results, it is concluded that C. villosus CTS8 and CTS57 strains represent a new lineage within the Bradyrhizobium genus.

Published
2014
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