Aboubacry Kane

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.

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.

Phylogeny of Nodulation Genes and Symbiotic Diversity of "Acacia senegal" (L.) Willd. and "A. seyal" (Del.) "Mesorhizobium" Strains from Different Regions of Senegal

Acacia senegal and Acacia seyal are small, deciduous legume trees, most highly valued for nitrogen fixation and for the production of gum arabic, a commodity of international trade since ancient times. Symbiotic nitrogen fixation by legumes represents the main natural input of atmospheric N2 into ecosystems which may ultimately benefit all organisms. We analyzed the nod and nif symbiotic genes and symbiotic properties of root-nodulating bacteria isolated from A. senegal and A. seyal in Senegal. The symbiotic genes of rhizobial strains from the two Acacia species were closed to those of Mesorhizobium plurifarium and grouped separately in the phylogenetic trees. Phylogeny of rhizobial nitrogen fixation gene nifH was similar to those of nodulation genes (nodA and nodC). All A. senegal rhizobial strains showed identical nodA, nodC, and nifH gene sequences. By contrast, A. seyal rhizobial strains exhibited different symbiotic gene sequences. Efficiency tests demonstrated that inoculation of both Acacia species significantly affected nodulation, total dry weight, acetylene reduction activity (ARA), and specific acetylene reduction activity (SARA) of plants. However, these cross-inoculation tests did not show any specificity of Mesorhizobium strains toward a given Acacia host species in terms of infectivity and efficiency as stated by principal component analysis (PCA). This study demonstrates that large-scale inoculation of A. senegal and A. seyal in the framework of reafforestation programs requires a preliminary step of rhizobial strain selection for both Acacia species.

Field Trials Reveal Ecotype-Specific Responses to Mycorrhizal Inoculation in Rice.

The overuse of agricultural chemicals such as fertilizer and pesticides aimed at increasing crop yield results in environmental damage, particularly in the Sahelian zone where soils are fragile. Crop inoculation with beneficial soil microbes appears as a good alternative for reducing agricultural chemical needs, especially for small farmers. This, however, requires selecting optimal combinations of crop varieties and beneficial microbes tested in field conditions. In this study, we investigated the response of rice plants to inoculation with arbuscular mycorrhizal fungi (AMF) and plant growth promoting bacteria (PGPB) under screenhouse and field conditions in two consecutive seasons in Senegal. Evaluation of single and mixed inoculations with AMF and PGPB was conducted on rice (Oryza sativa) variety Sahel 202, on sterile soil under screenhouse conditions. We observed that inoculated plants, especially plants treated with AMF, grew taller, matured earlier and had higher grain yield than the non-inoculated plants. Mixed inoculation trials with two AMF strains were then conducted under irrigated field conditions with four O. sativa varieties, two O. glaberrima varieties and two interspecific NERICA varieties, belonging to 3 ecotypes (upland, irrigated, and rainfed lowland). We observed that the upland varieties had the best responses to inoculation, especially with regards to grain yield, harvest index and spikelet fertility. These results show the potential of using AMF to improve rice production with less chemical fertilizers and present new opportunities for the genetic improvement in rice to transfer the ability of forming beneficial rice-microbe associations into high yielding varieties in order to increase further rice yield potentials.

Arbuscular mycorrhizal fungi (AMF) communities associated with cowpea in two ecological site conditions in Senegal

The objective of this study was to characterize the diversity of arbuscular mycorrhizal fungal (AMF) communities colonizing the roots of Vigna unguiculata (L.) plants cultivated in two different sites in Senegal. Roots of cowpea plants and soil samples were collected from two fields (Ngothie and Diokoul) in the rural community of Dya (Senegal). Microscopic observations of the stained roots indicated a high colonization rate in roots from Ngothie site as compared to those from Diokoul site. The partial small subunit of ribosomal DNA genes was amplified from the genomic DNA extracted from these roots by polymerase chain reaction (PCR) with the universal primer NS31 and a fungal-specific primer AML2. Nucleotide sequence analysis revealed that 22 sequences from Ngothie site and only four sequences from Diokoul site were close to those of known arbuscular mycorrhizal fungi. Also, 47.6% of the clones from Ngothie site and 89.47% from Diokoul site were not close to known AMF. A total of 15 operational taxonomic units (OUT) were identified. Phylogenetic analyses showed that these clones belonged to the genera Glomus, Sclerocystis, Rhizophagus, Scutellospora, Gigaspora, Racocetra, Acaulospora and Redeckera. The genus Glomus is the most represented with six OTU, representing 40% of all OTU. Key words: Agriculture, Glomeromycota, Vigna unguiculata, diversity, soil origin.

The rhizosphere of the halophytic grass Sporobolus robustus Kunth hosts rhizobium genospecies that are efficient on Prosopis juliflora (Sw.) DC and Vachellia seyal (Del.) seedlings

The aim of this study was to survey the abundance and genetic diversity of legume-nodulating rhizobia (LNR) in the rhizosphere of a salt-tolerant grass, Sporobolus robustus Kunth, in the dry and rainy seasons along a salinity gradient, and to test their effectiveness on Prosopis juliflora (SW.) DC and Vachellia seyal (Del.) P.J.H. Hurter seedlings. The results showed a significant decrease in LNR population density and diversity in response to salinity, particularly during the dry season. A phylogenetic analysis of the 16S-23S rRNA ITS region clustered the 232 rhizobium isolates into three genera and 12 distinct representative genotypes: Mesorhizobium (8 genotypes), Ensifer (2 genotypes) and Rhizobium (2 genotypes). Of these genotypes, 2 were only found in the dry season, 4 exclusively in the rainy season and 6 were found in both seasons. Isolates of the Mesorhizobium and Ensifer genera were more abundant than those of Rhizobium, with 55%, 44% and 1% of the total strains, respectively. The abundance of the Mesorhizobium isolates appeared to increase in the dry season, suggesting that they were more adapted to environmental aridity than Ensifer genospecies. Conversely, Ensifer genospecies were more tolerant of high salinity levels than the other genospecies. However, Ensifer genospeciesproved to be the most efficient strains on P. juliflora and V. seyal seedlings. We concluded that S. robustus hosts efficient rhizobium strains in its rhizosphere, suggesting its ability to act as a nurse plant to facilitate seedling recruitment of P. juliflora and V. seyal in saline soils.

Growth Response of Different Species and Provenances of Jujube Seedlings to Inoculation with Arbuscular Mycorrhizal Fungi

Many species of Ziziphus are underutilized crops despite their potential usage in agroforestry systems. The aim of our study was to evaluate the effects of arbuscular mycorrhizal fungi (AMF) on the growth and mineral nutrition of Ziziphus spp. and provenances of Z. mauritiana in greenhouse conditions. Three isolates of AMF were used: Rhizophagus irregularis IR27, Funneliformis mosseae DAOM227131, and Rhizophagus irregularis DAOM197198. They were inoculated on seven species of Ziziphus (Z. mauritiana, Z. lotus, Z. spina-christi, Z. mucronata, Z. amphibia, Z. abyssinica, and Z. sphaerocarpa) and six provenances of Z. mauritiana (Senegal, Mali, Mauritania, Burkina Faso, Niger, and India). Plants were grown in nursery receiving 20 g of crude inoculum of AMF. Two greenhouse experiments were set up: (1) a 4×6 factorial design consisting of three AMF, one control (disinfected soil without inoculum), and six provenances of Z. mauritiana and (2) a 4×7 factorial design consisting of three AMF, one control and seven Ziziphus spp. Inoculation by AMF significantly improved growth and mineral nutrition of Ziziphus spp., particularly P nutrition. All the Ziziphus spp. (except for Z. lotus) and provenances of jujube seedlings showed the highest mycorrhizal dependency (MD) values when inoculated with R. irregularis IR27. The differences of MD among the tested Ziziphus spp. seem to be due to the differences in the development of hyphal length in the soil and in P uptake by the external hyphae. Rhizophagus irregularis IR27 constitutes a promising biological tool for the production of higher-quality nursery stock with expected improved performance of Ziziphus spp. and Z. mauritiana provenances in agroforestry systems.

Effect of native and allochthonous arbuscular mycorrhizal fungi on Casuarina equisetifolia growth and its root bacterial community

ABSTRACT Exotic trees are often planted to recover degraded lands. Inoculation with mycorrhizal fungi can improve their survival. Plant growth is partly dependent on the strain used, but little attention has been paid to the selection of mycorrhizal fungi. The aim of this study was to determine whether the growth of Casuarina equisetifolia L. (Johnson) is affected by two different mycorrhizal inocula generated using fungal spores retrieved from an Australian site (allochthonous soil) and a Senegalese site (native soil) under C. equisetifolia trees. Comparative experiments were conducted with plants in a Senegalese soil, previously sterilized or not, and grown in a greenhouse. At harvest, parameters related to plant growth and mycorrhization were evaluated and soil bacterial communities were compared. Tree growth was significantly influenced by both types of inoculants. In unsterilized soil, plants inoculated with the native inoculant were taller than plants inoculated with the allochthonous inoculant and control plants. The frequency of mycorrhization with both inoculants was higher in unsterilized soil. The strongest effects of the mycorhizosphere on the soil microbiome were obtained with the allochthonous inoculum, and analysis of the taxonomic composition revealed mycorrhizal communities specific to each inoculum. These results suggest that the development of C. equisetifolia and its root bacterial community are dependent on the composition of the mycorrhizal inoculum. The functional consequences of this rhizosphere effect in terms of soil fertility should be further studied to better guide reforestation operations.

Growth and physiological responses of Sporobolus robustus kunth seedlings to salt stress

ABSTRACT Seedlings of S. robustus were exposed to increasing NaCl concentrations (0, 50, 100, 150, 200, 250, and 300 mM) for 50, 100, and 150 days, in greenhouse conditions. Total dry weight and salt tolerance index decreased gradually with increasing NaCl concentrations. The optimum growth of S. robustus (4.12 to 5.25 g · plant−1) was obtained between 0 and 150 mM at 150 days after salt stress. Foliar chlorophyll a and total chlorophyll contents increased with NaCl concentration at 50 days after treatment. There was no significant effect of salinity on chlorophyll a, b and total chlorophyll contents at 100 and 150 days after treatment. Higher Na contents were found in the shoots as compared to the roots. The Na content increased, while K decreased with increasing NaCl concentrations, suggesting competitive inhibition between absorptions of Na and K as a consequence, the K/Na ratios in shoots and roots decreased with increasing salinity. The proline contents in S. robustus were more pronounced at 300 mM (2.02 µmol/g), 250 mM (2.64 µmol/g), and 200 mM NaCl concentrations (2.98 µmol/g) for 50, 100, and 150 days, respectively, as compared to the treatment without added NaCl. Overall, S. robustus could be considered as salt tolerant on the basis of their performance in biomass production, accumulation of Na, similar foliar chlorophyll a, chlorophyll b and total chlorophyll contents, and accumulation of proline with increasing salinity. The potential ability of S. robustus to accumulate significant amounts of Na makes this halophyte promising as a desalinization tool of salted soils.