Isolation of mutants of the nitrogen-fixing actinomycete Frankia.

Frankia is a nitrogen (N)-fixing multicellular actinomycete which establishes root-nodule symbiosis with actinorhizal plants. Several aspects of Frankia N fixation and symbiosis are distinct, but genes involved in the specific features are largely unknown because of the lack of an efficient mutant screening method. In this study, we isolated mutants of Frankia sp.

Codon-optimized antibiotic resistance gene improves efficiency of transient transformation in Frankia.

Frankia is a unique actinobacterium having abilities to fix atmospheric dinitrogen and to establish endosymbiosis with trees, but molecular bases underlying these interesting characteristics are poorly understood because of a lack of stable transformation system. Extremely high GC content of Frankia genome (more than 70 percent) can be a hindrance to successful transformation. We generated a synthetic gentamicin resistance gene whose codon usage is optimized to Frankia (fgmR) and evaluated its usefulness as a selection marker using a transient transformation system.

Characteristics of bacteroids in indeterminate nodules of the leguminous tree Leucaena glauca.

Rhizobia establish symbiosis with legumes. Bacteroids in indeterminate nodules of Inverted Repeat Lacking Clade (IRLC) legumes undergo terminal differentiation caused by Nodule-specific Cysteine-Rich peptides (NCRs). Microscopic observations of bacteroids and the detection of NCRs in indeterminate nodules of the non-IRLC legume Leucaena glauca were performed.

The Frankia alni symbiotic transcriptome.

The actinobacteria Frankia spp. are able to induce the formation of nodules on the roots of a large spectrum of actinorhizal plants, where they convert dinitrogen to ammonia in exchange for plant photosynthates. In the present study, transcriptional analyses were performed on nitrogen-replete free-living Frankia alni cells and on Alnus glutinosa nodule bacteria, using whole-genome microarrays.

Identification by suppression subtractive hybridization of Frankia genes induced under nitrogen-fixing conditions.

Frankia is an actinobacterium that fixes nitrogen under both symbiotic and free-living conditions. We identified genes upregulated in free-living nitrogen-fixing cells by using suppression subtractive hybridization. They included genes with predicted functions related to nitrogen fixation, as well as with unknown function.

Transient transformation of frankia by fusion marker genes in liquid culture.

Frankia is a nitrogen-fixing actinobacterium that establishes root nodule symbiosis with actinorhizal plants. The molecular basis of the symbiosis is largely unknown because genetic manipulation of Frankia has not been feasible. In this study we made novel technical attempts to transform Frankia strain CcI3.