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

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 200

Composite Actinorhizal Plants with Transgenic Roots for the Study of Symbiotic Associations with Frankia

More than 200 species of dicotyledonous plants belonging to eight different families and 24 genera can establish actinorhizal symbiosis with the nitrogen-fixing soil actinomycete Frankia. Compared to the symbiotic interaction between legumes and rhizobia, little is known about the molecular basis of the infection process and nodule formation in actinorhizal plants. Here, we review a gene transfer system based on Agrobacterium rhizogenes that opens the possibility to rapidly analyze the function of candidate symbiotic genes. The transformation protocol generates ?composite plants? that consist of a nontransgenic aerial part with transformed hairy roots. Composite plants have already been obtained in three different species of actinorhizal plants, including the tropical tree species Casuarina glauca, the Patagonian shrub Discaria trinervis, and the nonwoody plant Datisca glomerata. The potential of this technique to advancing our understanding of the molecular mechanisms underlying infection by Frankia is demonstrated by functional analyses of symbiotic genes.Fil: Meriem Benabdoun, Faiza. Université Mentouri; ArgeliaFil: Nambiar Veetil, Mathish. No especifíca;Fil: Imanishi, Leandro Ezequiel. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Svistoonoff, Sergio. No especifíca;Fil: Ykhlef, Nadia. Université Mentouri; ArgeliaFil: Gherbi, Hassen. No especifíca;Fil: Franche, Claudine. No especifíca

Heart of endosymbioses : transcriptomics reveals a conserved genetic program among arbuscular mycorrhizal, actinorhizal and legume-rhizobial symbioses

To improve their nutrition, most plants associate with soil microorganisms, particularly fungi, to form mycorrhizae. A few lineages, including actinorhizal plants and legumes are also able to interact with nitrogen-fixing bacteria hosted intracellularly inside root nodules. Fossil and molecular data suggest that the molecular mechanisms involved in these root nodule symbioses (RNS) have been partially recycled from more ancient and widespread arbuscular mycorrhizal (AM) symbiosis. We used a comparative transcriptomics approach to identify genes involved in establishing these 3 endosymbioses and their functioning. We analysed global changes in gene expression in AM in the actinorhizal tree C. glauca. A comparison with genes induced in AM in Medicago truncatula and Oryza sativa revealed a common set of genes induced in AM. A comparison with genes induced in nitrogen-fixing nodules of C. glauca and M. truncatula also made it possible to define a common set of genes induced in these three endosymbioses. The existence of this core set of genes is in accordance with the proposed recycling of ancient AM genes for new functions related to nodulation in legumes and actinorhizal plants

Casuarina glauca prenodule cells display the same differentiation as the corresponding nodule cells

Recent phylogenetic studies have implied that all plants able to enter root nodule symbioses with nitrogen-fixing bacteria go back to a common ancestor (D.E. Soltis, P.S. Soltis, D.R. Morgan, S.M. Swensen, B.C. Mullin, J.M. Dowd, and P.G. Martin, Proc. Natl. Acad. Sci. USA, 92:2647-2651, 1995). However, nodules formed by plants from different groups are distinct in nodule organogenesis and structure. In most groups, nodule organogenesis involves the induction of cortical cell divisions. In legumes these divisions lead to the formation of a nodule primordium, while in non-legumes they lead to the formation of a so-called prenodule consisting of infected and uninfected cells. Nodule primordium formation does not involve prenodule cells, and the function of prenodules is not known. Here, we examine the differentiation of actinorhizal prenodule cells in comparison to nodule cells with regard to both symbionts. Our findings indicate that prenodules represent primitive symbiotic organs whose cell types display the same characteristics as their nodule counterparts. The results are discussed in the context of the evolution of root nodule symbiose