Contrasted evolutionary histories of two Toll-like receptors (Tlr4 and Tlr7) in wild rodents (MURINAE)

Background: In vertebrates, it has been repeatedly demonstrated that genes encoding proteins involved in pathogen-recognition by adaptive immunity (e. g. MHC) are subject to intensive diversifying selection. On the other hand, the role and the type of selection processes shaping the evolution of innate-immunity genes are currently far less clear. In this study we analysed the natural variation and the evolutionary processes acting on two genes involved in the innate-immunity recognition of Microbe-Associated Molecular Patterns (MAMPs). Results: We sequenced genes encoding Toll-like receptor 4 (Tlr4) and 7 (Tlr7), two of the key bacterial-and viral-sensing receptors of innate immunity, across 23 species within the subfamily Murinae. Although we have shown that the phylogeny of both Tlr genes is largely congruent with the phylogeny of rodents based on a comparably sized non-immune sequence dataset, we also identified several potentially important discrepancies. The sequence analyses revealed that major parts of both Tlrs are evolving under strong purifying selection, likely due to functional constraints. Yet, also several signatures of positive selection have been found in both genes, with more intense signal in the bacterial-sensing Tlr4 than in the viral-sensing Tlr7. 92% and 100% of sites evolving under positive selection in Tlr4 and Tlr7, respectively, were located in the extracellular domain. Directly in the Ligand-Binding Region (LBR) of TLR4 we identified two rapidly evolving amino acid residues and one site under positive selection, all three likely involved in species-specific recognition of lipopolysaccharide of gram-negative bacteria. In contrast, all putative sites of LBRTLR7 involved in the detection of viral nucleic acids were highly conserved across rodents. Interspecific differences in the predicted 3D-structure of the LBR of both Tlrs were not related to phylogenetic history, while analyses of protein charges clearly discriminated Rattini and Murini clades. Conclusions: In consequence of the constraints given by the receptor protein function purifying selection has been a dominant force in evolution of Tlrs. Nevertheless, our results show that episodic diversifying parasite-mediated selection has shaped the present species-specific variability in rodent Tlrs. The intensity of diversifying selection was higher in Tlr4 than in Tlr7, presumably due to structural properties of their ligands

Analysis of the Cultivable Endophytic Bacterial Diversity in the Date Palm (Phoenix dactylifera L.) and Evaluation of Its Antagonistic Potential against Pathogenic Fusarium Species that Cause Date Palm Bayound DiseaseAnalysis of the Cultivable Endophytic

Abstract Biological control still remains an unexploited issue in southern countries such as Tunisia. Thus, the present study focused on the diversity of cultivable endophytic bacteria in the internal tissues (roots and leaves) of Tunisian date palm trees (Phoenix dactylifera L.). In order to assess their antagonistic potential towards date palm pathogens, particularly Fusarium. Indeed, the Genus Fusarium includes the causative agent of the Bayound disease, Fusarium oxysporum, a major treat for date production North Africa. Twenty two bacterial isolates presenting distinct colony morphology on TSA media were selected. The latter were characterized using Gram staining, biochemical tests, and molecular identification techniques based on 16S rRNA gene sequencing. Cultivable endophytic isolates were assigned into seven distinct groups. The species Arthrobacter agilis and Bacillus subtilis exhibited lasting antagonistic properties against a range of Fusarium species including the causing agent of the Bayoud disease, thus demonstrating their strong potential for future applications in the inoculation of date palm trees for biocontrol purposes. The isolates showed extracellular enzymatic activity including cellulase (76, 92%), protease (69, 23%) and amylase (38, 46%). This study thus demonstrates for the first time that the diversity of endophytic bacteria is abundant in date palm trees (Phoenix dactylifera L.) and could present varying biotechnological applications and particularly disease control