Mame Ourèye Sy

Actinorhizal nitrogen fixing nodules: infection process, molecular biology and genomics

Actinorhizal hosts are non-leguminous perennial plants belonging to 8 angiosperm families. They are capable of forming root nodules as a result of infection by a nitrogen-fixing actinomycete called Frankia. Actinorhizal nodules consist of multiple lobes, each of which represents a modified lateral root with infected cells in the expanded cortex. This article summarizes the most recent knowledge about this original symbiotic process. The infection process is described both at cytological and molecular levels. The use of transgenic Casuarinaceae for studying in actinorhizal nodules the regulation of several symbiotic promoters from legumes is also discussed. With progress in plant genome sequencing, comparative genomics in legumes and actinorhizal plants should contribute to the understanding of the evolutionary history of nitrogen-fixing symbioses. Key words : Nitrogen-fixation, actinorhizal nodules, Frankia, Casuarina, symbiotic gene. African Journal of Biotechnology Vol. 2 (12), pp. 528-538, December 2003

Tolerance to environmental stress by the nitrogen-fixing actinobacterium Frankia and its role in actinorhizal plants adaptation

Environmental stresses are caused by human activities or natural events. Several of them including salinity, heavy metals, and extreme temperature affect both soil characteristics and plant growth and productivity. Actinorhizal plants are pioneer species that are able to grow in poor soils and improve soil fertility. They are widely used in agroforestry for different purposes including reclamation of degraded and contaminated lands. This capacity is mainly due to the plants forming a nitrogen-fixing symbiosis with actinobacteria known as Frankia. In comparison to uninoculated plants, plants in symbiosis with Frankia have significantly improved plant growth, total biomass, and nitrogen and chlorophyll content which enhance the development of actinorhizal plants and their resistance to abiotic stresses. However, to optimize the adaptation of actinorhizal species to different environments, selection of both symbiotic partners is necessary. Frankia strains vary in their sensitivity and response to stress including salinity, heavy metals, extreme pH and drought. In this paper, we review the response of different Frankia strains to environmental stresses and their role that they play in the adaptation of actinorhizal plants to stressful conditions.

NaCl Effects on In Vitro Germination and Growth of Some Senegalese Cowpea (Vigna unguiculata (L.) Walp.) Cultivars

Cowpea (Vigna unguiculata (L.) Walp.) is one of the most important grain legumes in sub-Saharian regions. It contributes to man food security by providing a protein-rich diet. However, its production is limited by abiotic stresses such as salinity. This study aims to evaluate the salt tolerance of 15 cowpea cultivars, at germination stage. The seed germination process consisted of sowing them in agarified water (8 g·L−1) supplemented with 6 different concentrations of NaCl (0, 10, 50, 100, 150, and 200 mM). Results highlighted that high salt concentrations drastically reduced germination and significantly delayed the process for all varieties. A cowpea varietal effect towards the salt tolerance was noticed. Genotypes Diongoma, 58-78, and 58-191 were more salt-tolerant cultivars while Mougne and Yacine were more salt-sensitive ones as confirmed in the three groups of the dendrogram. NaCl effects on the early vegetative growth of seedlings were assessed with a tolerant (58-191) and a susceptible (Yacine) cultivar. Morphological (length and dry biomass) and physiological (chlorophyll and proline contents) parameter measurements revealed a negative effect of high (NaCl). However, 58-191 was much more salt tolerant, and the chlorophyll and proline contents were higher than those of Yacine genotype at increasing salt concentrations.

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.

Permanent Draft Genome Sequence for Frankia sp. Strain CeD, a Nitrogen-Fixing Actinobacterium Isolated from the Root Nodules of Casuarina equistifolia Grown in Senegal

ABSTRACT Frankia strain CeD is a member of Frankia lineage Ib that is able to reinfect plants of the Casuarina families. Here, we report a 5.0-Mbp draft genome sequence with a G+C content of 70.1% and 3,847 candidate protein-encoding genes.

Molecular Biology of Tropical Nitrogen-Fixing Trees in the Casuarinaceae Family

The Casuarinaceae family includes about eighty species of shrubs and trees belonging to four genera, namely: Allocasuarina, Casuarina, Ceuthostoma, and Gymnostoma. Casuarinaceae are primarily native to the Southern Hemisphere, mostly Australia and Indo-Pacific areas, from Malaysia to Polynesia. However, the range of distribution of some genera such as Casuarina has been extended considerably through artificial dissemination. All members of the family are characterized by highly reduced leaves and photosynthetic deciduous branchlets (Midgley et al., 1983; National Research Council, 1984).

Symbiotic Performance of Diverse Frankia Strains on Salt-Stressed Casuarina glauca and Casuarina equisetifolia Plants

Symbiotic nitrogen-fixing associations between Casuarina trees and the actinobacteria Frankia are widely used in agroforestry in particular for salinized land reclamation. The aim of this study was to analyze the effects of salinity on the establishment of the actinorhizal symbiosis between C. glauca and two contrasting Frankia strains (salt sensitive; CcI3 vs. salt tolerant; CeD) and the role of these isolates in the salt tolerance of C. glauca and C. equisetifolia plants. We show that the number of root nodules decreased with increasing salinity levels in both plants inoculated with CcI3 and CeD. Nodule formation did not occur in seedlings inoculated with CcI3 and CeD, at NaCl concentrations above 100 and 200 mM, respectively. Salinity also affected the early deformation of plant root hairs and reduced their number and size. In addition, expression of symbiotic marker Cg12 gene, which codes for a subtilase, was reduced at 50 mM NaCl. These data suggest that the reduction of nodulation in C. glauca under salt stress is in part due to inhibition of early mechanisms of infection. We also show that prior inoculation of C. glauca and C. equisetifolia with Frankia strains CcI3 and CeD significantly improved plant height, dry biomass, chlorophyll and proline contents at all levels of salinity tested, depending on the Casuarina-Frankia association. There was no correlation between in vitro salt tolerance of Frankia strains and efficiency in planta under salt-stressed conditions. Our results strongly indicate that increased N nutrition, photosynthesis potential and proline accumulation are important factors responsible for salt tolerance of nodulated C. glauca and C. equisetifolia.

Symbiotic ability of diverse Frankia strains on Casuarina glauca plants in hydroponic conditions

Frankia is a soil actinomycete that forms nitrogen-fixing root nodule symbioses with eight angiosperms families including Casuarinaceae. Knowledge on symbiotic performance of several isolated strains with Casuarina species is limited. In this study, we characterized a collection of Frankia strains based on their growth kinetics and their symbiotic ability with Casuarina glauca specie. Results showed that Frankia strains Allo2, CcI3, CeD and Cg70.9 do not exhibit the same symbiotic ability both for their infectivity and effectiveness towards Casuarina glauca. All strains were able to infect and improve C. glauca plants growth. Frankia isolate CcI3 better improved plants height while CeD and Cg70.9 strains formed more nodules on inoculated plants roots. However, there is no correlation between the number of nodules formed and the effectiveness of strains. These preliminary results give ideas on which Frankia strains to use for obtaining an efficient symbiosis with C. glauca. However, others plant species and growth conditions should be tested for a longer period to better optimize the use of Frankia in reforestation programs.

Permanent Draft Genome Sequence of Frankia sp. Strain Allo2, a Salt-Tolerant Nitrogen-Fixing Actinobacterium Isolated from the Root Nodules of Allocasuarina

ABSTRACT Frankia sp. strain Allo2 is a member of Frankia lineage Ib, which is able to reinfect plants of the Casuarinaceae family, and exhibits a high level of salt tolerance compared to other isolates. Here, we report the 5.3-Mbp draft genome sequence of Frankia sp. strain Allo2 with a G+C content of 70.0% and 4,224 candidate protein-encoding genes.

Remediation of Heavy Metal-Contaminated Soils and Enhancement of Their Fertility with Actinorhizal Plants

Heavy metal-contaminated soils are a major concern for worldwide agricultural production. The main sources of environment pollution by heavy metal are the rapid urbanization and/or industrial growth. These pollutants induce the loss of the structure and the function of soil and microbial community. They affect human health through food chain and water contamination. To rehabilitate lands degraded by heavy metal, some efficiency and low-cost approaches have to be promoted widely. Phytoremediation is a potential tool to clean up pollutants from the contaminated site, implement a ground cover, and improve soil fertility. It is well known that symbiotic microorganisms tolerant to heavy metal increase plant resistance and toxic metal alleviation. So, management of the relationship between hyper-accumulator plants and these microorganisms increases the potential of this biological tool. In this chapter, we discuss (1) the use of actinorhizal plants for remediation of heavy metal-contaminated soils, (2) the impact of mycorrhizal fungi and/or nitrogen-fixing bacteria on plant adaptation and performance in heavy metal-contaminated sites, and (3) the positive effect of actinorhizal plants on soil fertility of heavy metal-contaminated soil.