Diégane Diouf

Hairy root nodulation of Casuarina glauca: a system for the study of symbiotic gene expression in an actinorhizal tree.

The purpose of this study was to establish a fast system for producing transgenic actinorhizal root nodules of Casuarina glauca. Agrobacterium rhizogenes strain A4RS carrying the p35S-gusA-int gene construct was used to induce hairy roots on hypocotyls of 3-week-old C. glauca seedlings. Three weeks after wounding, the original root system was excised, and composite plants consisting of transgenic roots on untransformed shoots were transferred to test tubes to be inoculated with Frankia. The actinorhizal nodules formed on transformed roots had the nitrogenase activity and morphology of untransformed nodules. beta-Glucuronidase (GUS) activity was examined in transgenic roots and nodules by fluorometric and histochemical assays. The results indicate that transgenic nodules generated with this root transformation system could facilitate the molecular study of symbiotic nitrogen fixation in actinorhizal trees.

Genetic Diversity of Acacia seyal Del. Rhizobial Populations Indigenous to Senegalese Soils in Relation to Salinity and pH of the Sampling Sites

The occurrence and the distribution of rhizobial populations naturally associated to Acacia seyal Del. were characterized in 42 soils from Senegal. The diversity of rhizobial genotypes, as characterized by polymerase chain reaction restriction fragment length polymorphism (RFLP) analysis of 16S–23S rDNA, performed on DNA extracted from 138 nodules resulted in 15 clusters. Results indicated the widespread occurrence of compatible rhizobia associated to A. seyal in various ecogeographic areas. However, the clustering of rhizobial populations based on intergenic spacer (IGS) RFLP profiles did not reflect their geographic origin. Four genera were discriminated on the basis of 16S rRNA gene sequences of the strains representative for the IGS-RFLP profiles. The majority of rhizobia associated to A. seyal were affiliated to Mesorhizobium and Sinorhizobium 64 and 29%, respectively, of the different IGS-RFLP profiles. Our results demonstrate the coexistence inside the nodule of plant-pathogenic non-N2-fixing Agrobacterium and Burkholderia strains, which induced the formation of ineffective nodules, with symbiotic rhizobia. Nodulation was recorded in saline soils and/or at low pH values or in alkaline soils, suggesting adaptability of natural rhizobial populations to major ecological environmental stress and their ability to establish symbiotic associations within these soil environments. These results contribute to the progressing research efforts to uncover the biodiversity of rhizobia and to improve nitrogen fixation in agroforestry systems in sub-Saharan Africa.

Symbiosis of Acacia auriculiformis and Acacia mangium with mycorrhizal fungi and Bradyrhizobium spp. improves salt tolerance in greenhouse conditions

The aim of our work was to assess the growth and mineral nutrition of salt stressed Acacia auriculiformis A. Cunn. ex Benth. and Acacia mangium Willd. seedlings inoculated with a combination of selected microsymbionts (bradyrhizobia and mycorrhizal fungi). Plants were grown in greenhouse conditions in non-sterile soil, irrigated with a saline nutrient solution (0, 50 and 100 mm NaCl). The inoculation combinations consisted of the Bradyrhizobium strain Aust 13c for A. mangium and Aust 11c for A. auriculiformis, an arbuscular mycorrhizal fungus (Glomus intraradices, DAOM 181602) and an ectomycorrhizal fungus (Pisolithus albus, strain COI 007). The inoculation treatments were designed to identify the symbionts that might improve the salt tolerance of both Acacia species. The main effect of salinity was reduced tree growth in both acacias. However, it appeared that, compared with controls, both rhizobial and mycorrhizal inoculation improved the growth of the salt-stressed plants, while inoculation with the ectomycorrhizal fungus strain appeared to have a small effect on their growth and mineral nutrition levels. Endomycorrhizal inoculation combined with rhizobial inoculation usually gave good results. Analysis of foliar proline accumulation confirmed that dual inoculation gave the trees better tolerance to salt stress and suggested that the use of this dual inoculum might be beneficial for inoculation of both Acacia species in soils with moderate salt constraints.

Phenotypic and genotypic characteristics of Acacia senegal (L.) Willd. root‐nodulating bacteria isolated from soils in the dryland part of Senegal

Aims:  We characterized phenotypically and genotypically root‐nodulating bacteria associated with Acacia senegal (L.) Willd. isolated from the soils surrounding A. senegal trees in the dry land area of Senegal.

Casuarina in Africa: Distribution, role and importance of arbuscular mycorrhizal, ectomycorrhizal fungi and Frankia on plant development

Exotic trees were introduced in Africa to rehabilitate degraded ecosystems. Introduced species included several Australian species belonging to the Casuarinaceae family. Casuarinas trees grow very fast and are resistant to drought and high salinity. They are particularly well adapted to poor and disturbed soils thanks to their capacity to establish symbiotic associations with mycorrhizal fungi -both arbuscular and ectomycorrhizal- and with the nitrogen-fixing bacteria Frankia. These trees are now widely distributed in more than 20 African countries. Casuarina are mainly used in forestation programs to rehabilitate degraded or polluted sites, to stabilise sand dunes and to provide fuelwood and charcoal and thus contribute considerably to improving livelihoods and local economies. In this paper, we describe the geographical distribution of Casuarina in Africa, their economic and ecological value and the role of the symbiotic interactions between Casuarina, mycorrhizal fungi and Frankia.

Phylogenetic analyses of symbiotic genes and characterization of functional traits of Mesorhizobium spp. strains associated with the promiscuous species Acacia seyal Del.

Aims:  To assess the phenotypic, symbiotic and genotypic diversity scope of Mesorhizobium spp. strains associated with Acacia seyal (Del.) isolated from different agro‐ecological zones in Senegal, and uses of susceptible microbial inoculum in a reafforestation process.

Effect of distance and depth on microbial biomass and mineral nitrogen content under Acacia senegal (L.) Willd. trees

The relations between plants and soil biota involve positive and negative feedbacks between soil organisms, their chemical environment, and plants. Then, characterization of microbial community functioning is important to understand these relations. An experiment was conducted in a field system in the north of Senegal for two years (2005 and 2006) in order to investigate the effect of depth and distance from Acacia senegal tree stem on soil microbial biomass and inorganic-N content. Soils were sampled during dry season (April, T(0)) and wet season (August, T(1)) along transects (R(0), foot tree; R(/2,) approximately 0.50 m distance from the stem; and R, approximately 1 m distance from the stem) and at different layers: 0-25 cm, 25-50 cm and 50-75 cm of A. senegal trees rhizosphere. Total microbial biomass and inorganic-N content were negatively correlated to the distance from tree stem and the depth. The highest values of microbial biomass and mineral nitrogen were found at the foot tree (R(0)) and at 0-25 cm layer. Inorganic-N was mostly in nitrate form (NO(3)(-)) during the dry season. In contrast, during the wet season, inorganic-N was dominated by ammoniac form (NH(4)(+)). Soil total microbial biomass and inorganic-N (NH(4)(+)+NO(3)(-)) were negatively correlated. Our results suggest a positive influence of A. senegal rhizosphere on soil microbial biomass and inorganic-N content.

Analysis of Genetic Diversity and Population Structure of Sesame Accessions from Africa and Asia as Major Centers of Its Cultivation

Sesame is an important oil crop widely cultivated in Africa and Asia. Understanding the genetic diversity of accessions from these continents is critical to designing breeding methods and for additional collection of sesame germplasm. To determine the genetic diversity in relation to geographical regions, 96 sesame accessions collected from 22 countries distributed over six geographic regions in Africa and Asia were genotyped using 33 polymorphic SSR markers. Large genetic variability was found within the germplasm collection. The total number of alleles was 137, averaging 4.15 alleles per locus. The accessions from Asia displayed more diversity than those from Africa. Accessions from Southern Asia (SAs), Eastern Asia (EAs), and Western Africa (WAf) were highly diversified, while those from Western Asia (WAs), Northern Africa (NAf), and Southeastern Africa (SAf) had the lowest diversity. The analysis of molecular variance revealed that more than 44% of the genetic variance was due to diversity among geographic regions. Five subpopulations, including three in Asia and two in Africa, were cross-identified through phylogenetic, PCA, and STRUCTURE analyses. Most accessions clustered in the same population based on their geographical origins. Our results provide technical guidance for efficient management of sesame genetic resources in breeding programs and further collection of sesame germplasm from these different regions.

Changes in Land Use System and Environmental Factors Affect Arbuscular Mycorrhizal Fungal Density and Diversity, and Enzyme Activities in Rhizospheric Soils of Acacia senegal (L.) Willd.

The responses of the soil microbial community features associated to the legume tree Acacia senegal (L.) Willd. including both arbuscular mycorrhizal fungal (AMF) diversity and soil bacterial functions, were investigated under contrasting environmental conditions. Soil samples were collected during dry and rainy seasons in two contrasting rainfall sites of Senegal (Dahra and Goudiry, in arid and semiarid zone, resp.). Soils were taken from the rhizosphere of A. senegal both in plantation and natural stands in comparison to bulk soil. A multiple analysis revealed positive correlations between soil physicochemical properties, mycorrhizal potential and enzyme activities variables. The positive effects of A. senegal trees on soil mycorrhizal potential and enzyme activities indicates that in sahelian regions, AMF spore density and diversity as well as soil microbial functions can be influenced by land-use systems (plantation versus natural population of A. senegal) and environmental conditions such as moisture and soil nutrient contents. Our study underlines the importance of prior natural AMF screening for better combinations of A. senegal seedlings with AMF species to achieve optimum plant growth improvement, and for restoration and reforestation of degraded lands.

Uvitex2B: a rapid and efficient stain for detection of arbuscular mycorrhizal fungi within plant roots

The study of arbuscular mycorrhiza often requires the staining of fungal structures using specific dyes. Fluorescent dyes such as acid fuchsin and wheat germ agglutinin conjugates give excellent results, but these compounds are either hazardous or very expensive. Here, we show that a safer and inexpensive dye, Uvitex2B, can be efficiently used to stain intraradical fungal structures formed by the arbuscular mycorrhizal fungus Glomus intraradices in three plant species: carrot, Casuarina equisetifolia, and Medicago truncatula. The intensity and stability of Uvitex2B allow the acquisition of high-quality images using not only confocal laser scanning microscopy but also epifluorescence microscopy coupled with image deconvolution. Furthermore, we demonstrate that Uvitex2B and β-glucuronidase staining are compatible and can thus be used to reveal arbuscular mycorrhizal structures in the context of promoter activation analysis.